CN102233077A

CN102233077A - Use of medicinal composition in preparation of medicine for improving activity of adenosine 5'-monophosphate-activated protein kinase (AMPK)

Info

Publication number: CN102233077A
Application number: CN2010101563320A
Authority: CN
Inventors: 仝小林; 甄重; 周水平
Original assignee: Tianjin Tasly Pharmaceutical Co Ltd
Current assignee: Tianjin Tasly Pharmaceutical Co Ltd
Priority date: 2010-04-27
Filing date: 2010-04-27
Publication date: 2011-11-09

Abstract

The invention belongs to the field of pharmacy and relates to the use of a medicinal composition in the preparation of a medicine for improving the activity of AMPK. The medicinal composition is prepared according to a formula consisting of the following Chinese medicinal raw materials in part by weight: 5 to 40 parts of mongolian snakegourd root, 10 to 30 parts of root of Chinese thorowax, 3 to 15 parts of immature bitter orange, 1 to 6 parts of raw rheum officinale, 1 to 12 parts of pinellia ternate, 3 to 15 parts of root of scutellaria baicalensis, 1 to 12 parts of coptis chinensis, 3 to 15 parts of white paeony root and 5 to 20 parts of dark plum.

Description

The application of a kind of pharmaceutical composition in the medicine of preparation raising AMPK enzymatic activity

Technical field

The invention belongs to pharmaceutical field, relate to the application of a kind of pharmaceutical composition in improving AMPK enzymatic activity medicine.

Background technology

Type 2 diabetes mellitus (T2DM) is the worldwide public health problem of serious threat human health.The statistical data that International Diabetes Federation (IDF) 2006 delivers shows that 20 years in the past, global diabetics number increased severely to 2.3 hundred million from 3,000 ten thousand.The expert of World Health Organization (WHO) estimates, will reach 3.5 hundred million to global diabetics in 2025.India, China, the U.S. are followed successively by three maximum countries of global diabetics quantity, estimate that Chinese diabetics total number of persons is nearly 4,000 ten thousand, are only second to India, occupy the second place of the world, account for 29.63% of the world.The control of T2DM foreseeable all will be the severe challenge that the mankind face in quite over a long time.

Type 2 diabetes mellitus also is adult's morbidity type diabetes, how to fall ill after 35～40 years old, accounts for diabetics more than 90%.The ability that produces insulin in the type 2 diabetes mellitus ward mate body is not to completely lose, insulin even produce too much in the patient's body that has, but the action effect of insulin has a greatly reduced quality, so the intravital insulin of patient may be in a kind of state of relative shortage.Can stimulate secretion of insulin in the body by some oral drugs.But still have the part patient need as type 1 diabetes, carry out insulinize to the later stage.Type 2 diabetes mellitus roughly has following 3 characteristics: first characteristics, type 2 diabetes mellitus is sent out in the adult well, is many with middle-aged and elderly people especially.Repeatedly diabetes epidemiological study result shows, the age of type 2 diabetes mellitus morbidity is many, and from 40 years old, prevalence increased gradually in 40-60 year, peaks to the geratic period.The maturity-onset diabetes of young morbidity comparatively speaking, and is still very rare.Second characteristic is generally relatively mitigations, hidden of the state of an illness, that is to say with type 1 diabetes and compares, and do not bear down menacingly so.The type 2 diabetes mellitus patient symptom is not obvious, may not be certain each patient all have drink manyly, urinate manyly, the performance of outeat, majority do not become thin significantly yet, to descend to some extent still be more common for muscle power and body weight certainly.The 3rd characteristics are exactly that the patient does not often need to earn a bare living by insulinize.That is to say that the patient does not beat insulin and is unlikely to ketoacidosis to take place soon and threat to life yet.So type 2 diabetes mellitus was non-insulin-dependent diabetes mellitus originally again.The type 2 diabetes mellitus patient also needs to make insulinize sometimes, but majority is because glycemic control is undesirable, or heavier because the chronic complicating diseases of acute complications or diabetes has taken place, and is in order to earn a bare living unlike the type 1 diabetes patient.

Type 2 diabetes mellitus is the disease more more complicated than type 1 diabetes, easier treatment when particularly the interior insulin of body still can self-ly produce in early days.Because early symptom slight (as no ketoacidosis and stupor etc.) and distributing, so diagnosed at the rear for many years at disease progression more than the type 2 diabetes mellitus more.Yet, the improper meeting of the control of type 2 diabetes mellitus cause such as renal failure, blind, wound healing slow, arteriopathy severe complications such as (comprising coronary artery disease).

Its principal character of type 2 diabetes mellitus patient is an insulin resistant, relatively insulin deficit and hyperglycemia; The increase of the hepatic glucose production degraded of hepatic glycogen (as derive from) is particularly in the increase (typical reason is the insulin level disorder, and this function of liver is regulated and control in one of effect of insulin just) on improper opportunity; The minimizing (pathway deficiency behind receptor and the receptor) of the glucose transport of insulin-mediated in muscle and the fatty tissue (mainly); The beta Cell of islet function is impaired---the function that early stage insulin discharges when having lacked the hyperglycemia levels of stimulation.

Chinese medicine can play more positive effect in treating diabetes." element is asked strange sick opinion ": " ... overflowing also of this five gas, name day the spleen-warm syndrome.Husband's five tastes inlet is hidden in stomach, and spleen is the Xingqi vital essence for it, and body fluid is at spleen, so make us sweet taste in the mouth also.This fertile institute is sent out also, and this person must count the sweet and refreshing and many fertilizer of food also, the overeating greasy food bringing about internal heat, the over-eating the food with sweet flavor bringing about abdominal distension is so its gas overflow transfers to and quenching one's thirst ".The Tong Xiaolin professor studies in conjunction with clinical epidemiology, proposition by soil stop up wood strongly fragrant due to the fullness in the epigastrium and abdomen interior-heat be (obesity) T2DM basic pathogenesis, the dissipating depression of QI heat clearing away is its basic method of treatment, through long-term clinical establishment dissipating depression of QI heat clearing away side's medicine and confirmed the clinical efficacy of dissipating depression of QI clearing away heat method, tentatively set forth its mechanism of action at integral level, histiocyte level and molecular level, comprise the dystopy deposition that alleviates triglyceride, improve receptor number and adhesion, influence insulin signaling transduction system etc.

The sugared quick spirit of dissipating depression of QI heat clearing away side medicine is a kind of traditional Chinese compound medicine for the treatment of diabetes, is made by Chinese medicines such as Radix Trichosanthis, Radix Bupleuri, Fructus Aurantii Immaturus, Radix Et Rhizoma Rhei.Through finding that with placebo the quick clever glycolated hemoglobin that reduces of sugar obviously is better than placebo, sugared quick spirit can not only significantly improve insulin resistant, also has the effect of significantly repairing and protect islet cell function.In addition, sugared quick spirit has good hypoglycemic effect to diabetics, for lipid metabolic disorder regulating action is preferably arranged.

Adenosine monophosphate activated protein kinase (AMPK) lifting effect in regulation of energy.The energy state of AMPK energy receptor cell, AMPK is activated when AMP/ATP ratio in the cell raises, energy metabolism is regulated in expression by phosphorylation effect protein or regulation and control range gene, close catabiotic metabolic pathway of synthesizing (as cholesterol, lipogenesis), activate the catabolic pathway (as fatty acid oxidation) that produces ATP, regulate downstream series factor rate-limiting enzyme activity such as SERBP-1, reduce the deposition of lipid, improve lipid metabolism in peripheral tissues.Simultaneously, the active reduction of AMPK is fat and former factor of type 2 diabetes mellitus Susceptible population generation insulin resistant, and hypothalamic AMPK enzymatic pathway also participates in trophic behavior, and therefore, AMPK is the crucial target spot of treatment fat toxicity, obesity and type 2 diabetes mellitus.

The present invention to sugar quick spirit done more deep research, find that unexpectedly sugared quick spirit can improve the AMPK enzymatic activity, for the therapeutic use of sugared Min Lingxin provides foundation.

Summary of the invention

The object of the invention is to provide the application of a kind of pharmaceutical composition in the medicine of preparation raising AMPK enzymatic activity.

Pharmaceutical composition of the present invention (is called the quick clever preparation of sugar again, the dissipating depression of QI fever-clearing preparation), by following parts by weight of Chinese traditional medicine crude drug is that prescription is formulated: Radix Trichosanthis 5-40 part, Radix Bupleuri 10-30 part, Fructus Aurantii Immaturus 3-15 part, Radix Et Rhizoma Rhei 1-6 part, Rhizoma Pinelliae 1-12 part, Radix Scutellariae 3-15 part, Rhizoma Coptidis 1-12 part, Radix Paeoniae Alba 3-15 part, Fructus Mume 5-20 part.

Fructus Crataegi in can also adding in the above-mentioned prescription, the prescription that obtains is: Radix Trichosanthis 10-30 part, Radix Bupleuri 10-30 part, Fructus Aurantii Immaturus 3-15 part, Radix Et Rhizoma Rhei 1-6 part, Rhizoma Pinelliae 1-12 part, Radix Scutellariae 3-15 part, Rhizoma Coptidis 1-12 part, Radix Paeoniae Alba 3-15 part, Fructus Mume 5-20 part, Fructus Crataegi 3-15 part.

Fructus Crataegi in can also adding in the above-mentioned prescription, Rhizoma Polygonati, Radix Ginseng and Fructus Momordicae charantiae, the prescription that obtains is: Radix Trichosanthis 10-30 part, Radix Bupleuri 10-30 part, Fructus Aurantii Immaturus 3-15 part, Radix Et Rhizoma Rhei, 1-6 part, Rhizoma Pinelliae 1-12 part, Radix Scutellariae 3-15 part, Rhizoma Coptidis 1-12 part, Radix Paeoniae Alba 3-15 part, Fructus Mume 5-20 part, Fructus Crataegi 3-15 part, Rhizoma Polygonati 3-15 part, Radix Ginseng 3-15 part, Fructus Momordicae charantiae 3-15 part.

Preferred prescription of the present invention is: 9 parts of Radix Trichosanthis, 12 parts of Radix Bupleuri, 9 parts of Fructus Aurantii Immaturuss, 3 parts of Radix Et Rhizoma Rhei, 6 parts of the Rhizoma Pinelliaes, 9 parts of Radix Scutellariaes, 6 parts of Rhizoma Coptidis, 9 parts of the Radix Paeoniae Albas, 9 parts of Fructus Mumes.

Or:

30 parts of Radix Trichosanthis, 12 parts of Radix Bupleuri, 9 parts of Fructus Aurantii Immaturuss, 3 parts of Radix Et Rhizoma Rhei, 6 parts of the Rhizoma Pinelliaes, 9 parts of Radix Scutellariaes, 6 parts of Rhizoma Coptidis, 9 parts of the Radix Paeoniae Albas, 15 parts of Fructus Mumes, 9 parts of Fructus Crataegis.

Or:

30 parts of Radix Trichosanthis, 12 parts of Radix Bupleuri, 9 parts of Fructus Aurantii Immaturuss, 3 parts of Radix Et Rhizoma Rhei, 6 parts of the Rhizoma Pinelliaes, 9 parts of Radix Scutellariaes, 6 parts of Rhizoma Coptidis, 9 parts of the Radix Paeoniae Albas, 15 parts of Fructus Mumes, 9 parts of Fructus Crataegis, 9 parts of Rhizoma Polygonatis, 9 parts of Radix Ginsengs, 9 parts in Fructus Momordicae charantiae.

In more than forming, weight is calculated with crude drug, and part is a weight portion, if with the gram is unit, more than composition can be made into a pharmaceutical preparation 5-50 preparation unit, and described preparation unit refers to, the final drug preparation of making, as make solid preparation 5-50 unit, oral liquid 5-50ml etc.

More than form the preparation that can be made into 1-6 taking dose, as tablet, make 18, each taking dose can be the 3-18 sheet, can take 1-6 time altogether.As granule, make 6 bags, take the 1-2 bag at every turn, can take 3-6 time altogether.

More than form to be by weight as proportioning, when producing, can increase or reduce according to corresponding proportion, as large-scale production can be unit with the kilogram, or be unit with the ton, small-scale production can be unit with the milligram also, weight can increase or reduce, but the constant rate of the raw medicinal herbs weight proportion between each composition.

The ratio of above weight proportion obtains through science screening, for especial patient, and as serious symptom or light disease, fat or modest patient, the proportioning of the amount of can corresponding adjustment forming increases or reduces being no more than 300%, and drug effect is constant.

Single medicinal material, especially ministerial drug and adjuvant drug in more than forming also can be replaced by the suitable Chinese medicine with identical property of medicine, and its drug effect of the Chinese medicine preparation after the replacement is constant.

Chinese medicine composition of the present invention is to process through extraction or other modes by the raw material of Chinese medicine that above-mentioned prescription is formed, and makes pharmaceutically active substance, subsequently, with this material is raw material, adds the medicine acceptable carrier when needing, and makes according to the routine techniques of galenic pharmacy.Described active substance can obtain by extracting raw material of Chinese medicine respectively, also can obtain by the co-extracted raw material of Chinese medicine, also can obtain by other modes, as: by pulverize, squeeze, calcine, grind, sieve, percolation, extraction, water are carried, alcohol extraction, ester are carried, methods such as ketone is carried, chromatography obtain, these active substances can be the material of extractum form, can be that dry extract also can be a fluid extract, make different concentration according to the different needs decision of preparation.

Pharmaceutically active substance in the Chinese medicine composition of the present invention, its shared percentage by weight in preparation can be 0.1-99.9%, all the other are the medicine acceptable carrier.Pharmaceutical composition of the present invention exists with unit dosage form, and described unit dosage form is meant the unit of preparation, as every of tablet, and capsular every capsules, every bottle of oral liquid, every bag of granule etc.

Chinese medicine composition of the present invention can be any pharmaceutically useful dosage form, and these dosage forms comprise: tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, oral liquid, suck agent, granule, electuary, pill, powder, unguentum, sublimed preparation, suspensoid, powder, solution, injection, suppository, ointment, plaster, cream, spray, drop, patch.Preparation of the present invention, peroral dosage form preferably, as: capsule, tablet, oral liquid, granule, pill, powder, sublimed preparation, unguentum etc.

Chinese medicine composition of the present invention, the preparation of its oral administration can contain excipient commonly used, such as binding agent, filler, diluent, tablet agent, lubricant, disintegrating agent, coloring agent, flavoring agent and wetting agent, can carry out coating to tablet in case of necessity.

The filler that is suitable for comprises cellulose, mannitol, lactose and other similar filler.Suitable disintegrating agent comprises starch, polyvinylpyrrolidone and starch derivatives, for example sodium starch glycollate.Suitable lubricant comprises, for example magnesium stearate.The acceptable wetting agent of appropriate drug comprises sodium lauryl sulphate.

Can fill by mixing, the method that tabletting etc. are commonly used prepares solid oral composition.Mix repeatedly active substance is distributed in those compositionss of a large amount of filleies of whole use.

The form of oral liquid for example can be aqueous or oily suspensions, solution, Emulsion, syrup or elixir, perhaps can be a kind of available water before use or other suitable composite dry products of carrier.This liquid preparation can contain conventional additive, such as suspending agent, for example sorbitol, syrup, methylcellulose, gelatin, hydroxyethyl-cellulose, carboxymethyl cellulose, aluminium stearate gel or hydrogenation edible fat, emulsifying agent, for example lecithin, anhydro sorbitol monooleate or arabic gum; Non-aqueous carrier (they can comprise edible oil), for example almond oil, fractionated coconut oil, such as oily ester, propylene glycol or the ethanol of the ester of glycerol; Antiseptic, for example para hydroxybenzene methyl ester or propyl p-hydroxybenzoate or sorbic acid, and if desired, can contain conventional flavouring agent or coloring agent.

For injection, the liquid unit dosage forms of preparation contains active substance of the present invention and sterile carrier.According to carrier and concentration, this chemical compound can be suspended or dissolving.The preparation of solution is normally by being dissolved in active substance in a kind of carrier filter-sterilized before it is packed into a kind of suitable bottle or ampoule, sealing then.For example a kind of local anesthetic of adjuvant, antiseptic and buffer agent also can be dissolved in this carrier.In order to improve its stability, can be after the bottle of packing into that this compositions is freezing, and under vacuum, water is removed.

Chinese medicine composition of the present invention, when being prepared into medicament, optionally add suitable medicine acceptable carrier, described medicine acceptable carrier is selected from: mannitol, sorbitol, sodium pyrosulfite, sodium sulfite, sodium thiosulfate, cysteine hydrochloride, TGA, methionine, vitamin C, the EDTA disodium, EDTA calcium sodium, the alkali-metal carbonate of monovalence, acetate, phosphate or its aqueous solution, hydrochloric acid, acetic acid, sulphuric acid, phosphoric acid, aminoacid, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivative, cellulose and derivant thereof, alginate, gelatin, polyvinylpyrrolidone, glycerol, soil temperature 80, agar, calcium carbonate, calcium bicarbonate, surfactant, Polyethylene Glycol, cyclodextrin, beta-schardinger dextrin-, the phospholipid material, Kaolin, Pulvis Talci, calcium stearate, magnesium stearate etc.

Compositions of the present invention is determined usage and dosage according to patient's situation in use, but obeys every day three times, each 1-20 agent, as: 1-20 bag or grain or sheet.

Compositions of the present invention is that concrete steps are as follows through following method preparation:

The preparation method of the quick clever preparation of sugar of the present invention is: (1) gets above-mentioned raw materials in proportion, and (2) add the ethanol of 4 times of amount 70-90%, 50-80 ℃ of decompression recycling ethanol, and 70-90 ℃ of vacuum drying pulverized 80 mesh sieves, got drug extract; Add adjuvant and make acceptable dosage form on any kind of pharmaceutics.

Pharmaceutical composition of the present invention and preparation method thereof can be with reference to Chinese patent CN1726929.

Other most preferred preparation methoies of the present invention in an embodiment.

Further specify the therapeutic effect of pharmaceutical composition of the present invention by following experiment.

Test a sugared quick spirit to spontaneous type 2 diabetes mellitus rat sugar, lipometabolic influence

1. materials and methods

1.1 laboratory animal

With spontaneous type 2 diabetes mellitus rat model OLETF Mus and its homology non-diabetic contrast Mus LETO Mus is experimental subject.Rat is all male, 40 of OLETF, and 10 of LETO rats all around age, are introduced by Japanese Otsuka Pharmaceutical Co., Ltd. Tokushima institute.

Rat is raised at no-special pathogen level (SPF level, specific pathogen-free) the condition cage that places an order, and raises with standard feed.Ambient temperature is controlled at 22-25 ℃, and humidity is 55 ± 5%, 12/12 hour light dark cycle (light application time 7:00-19:00), freely obtains food and drinking-water.

Raise the place: the Experimental Animal Center SPF of Institute of Radiation Medicine, Chinese Academy of Medical Sciences level animal feeding room.

1.2 grouping and administration

1.2.1OLETF diabetes diagnosis grouping standard

The regularly capable oral glucose tolerance test (OGTT) of rat.Test preceding fasting overnight 15 hours, can't help water, give 30% glucose solution by 2g/kg and irritate stomach (get glucose powder 82.5g, adding distil water is settled to 250ml, and heating for dissolving is limpid liquid, and obtaining concentration is 30% glucose solution).

In on an empty stomach and behind the glucose load 30,60,90 and 120min get blood, measure blood glucose value with the quick glucose analyser of card BIOSEN5030 in the Germany.Glycemic peaks＞16.7mmol/L and load back 120min blood glucose＞11.1mmol/L examine and are diabetes, possess an above-mentioned person and are impaired glucose tolerance.

1.2.2 laboratory animal grouping and administration

During to 30 weeks, have into 27 of mould OLETF Mus, be divided into model group, rosiglitazone group, the quick spirit group of sugar at random, LETO is the blank group.

Rosiglitazone group: rosiglitazone (Avandia): every of Smithkline Beecham's Tianjin company limited product contains rosiglitazone maleate 4mg, available from by U.S.'s GlaxoSmithKline PLC (China) Investment Co., Ltd, face with before being stirred to dissolving fully, with distilled water diluting, dosage is 3mg/kg/d ^-1

The quick spirit group of sugar (that is: dissipating depression of QI heat clearing away group) adopts the formulated of the embodiment of the invention 1 to become extractum, and 4 ℃ of refrigerators are preserved, and with distilled water diluting, irritating stomach dosage is 9mg/kg/d ^-1(calculating) by contained crude drug amount.

Model group is irritated stomach with blank group with the equivalent distilled water, and each organizes equal 12 weeks of gastric infusion, every day 1 time.

1.3 experimental technique

1.3.1 laboratory animal food ration

Weighing rat food ration is 1 time weekly.Throw the capacity feedstuff in advance for weekly rat, take by weighing surplus (the feedstuff surplus weighing time is all a whole morning 8 points) after the week.

1.3.2 experimental animals quality

The body constitution amount: claim Mus body constitution amount and record with the electronics weighing machine weekly, the time of taking by weighing is all a whole morning 9:00.

1.3.3 oral glucose tolerance test

Per 4 weeks carry out oral glucose tolerance test (OGTT) before the rat administration and after the administration.Fasting overnight 15 hours before the test, can't help water, give 30% glucose solution by 2g/kg and irritate stomach, on an empty stomach and behind the glucose load 30,60,90 and 120min cut tail and get blood, measure blood glucose.When 32 all oral carbohydrate tolerance were tested, inner canthus was got blood synchronously, measures insulin.

Blood glucose is measured by the quick glucose analyser of card BIOSEN5030 in the Germany.

Insulin assay is with putting the method for exempting from (with Linco company rat insulin medicinal box special, the special messenger is with batch mensuration).

Area calculates (AUC) under insulin, the blood glucose curve: adopt the computing formula of approximate trapezoid, 0-120min blood glucose AUC=1/2 (0min value+120min value)+30min value+60min value+90min value.

1.3.4 four of blood fat, free serum fatty acid, saccharifying serum albumin detect

Irritate after stomach finishes, rat fasting overnight 12 hours, inferior morning, the rat jugular vein was got blood 2.5ml, and centrifuging and taking serum is preserved to be equipped with for-70 ℃ and is surveyed.

Triglyceride (TG), T-CHOL (TC), low-density lipoprotein cholesterol (LDL-C), HDL-C (HDL-C).TC, LDL-C, HDL-C adopt enzyme process, and TG adopts the acetylacetone,2,4-pentanedione development process, and the transaminase carries out on Hitachi-7150 automatic clinical chemistry analyzer, is detected by the Gate of Pervasive Peace hospital laboratory; Saccharifying serum albumin BecKmanCX-5 automatic clinical chemistry analyzer is measured, and sample is with a collection of mensuration.

1.3.5 the positive glucose clamp test of hyperinsulinism

Rat fasting overnight, can't help water, pentobarbital sodium with 2% after weighing carries out intraperitoneal injection of anesthesia, dorsal position is fixed, after the anesthesia produce effects, cut skin of neck, passivity is separated subcutaneous tissue and muscle, separates to expose in right carotid and the left side neck or external jugular vein, carries out in right carotid and the left side neck or the external jugular vein intubate, the threading ligation, fixing (U.S. produces epidural anesthesia silica gel catheter, internal diameter 0.6mm, external diameter 1mm), and be full of conduit with the normal saline that contains heparin 50U/ml, keep it unobstructed.Arterial cannulation is surveyed blood glucose to get blood, and venous cannulation connects the mini three links pipe, and the port of export links to each other with vein, and two entrance points are connected with infusion of insulin and glucose with the digital micro-injection pump of micro computer respectively.After intubate finishes, rat left standstill measure blood glucose after 30 minutes.And simultaneously it is incubated.

Insulin, 20% glucose are pumped into by jugular vein with 2 digital micro-injection pumps of micro computer respectively, and blood preparation is obtained by carotid duct.The syringe that the intubate rat carotid artery conduit that left standstill 30 minutes is connect takes off, and gets one of blood, measures basic blood glucose value with miniature blood glucose meter in 30s.

(infusion rates is 8mU/kgmin to the fugitive Iletin II (Lilly) of elder generation's constant infusion during beginning ^-1, face the time spent insulin and dilute with 0.5% bovine serum albumin).Measured blood glucose once in per 10 minutes, when blood glucose value exceeds the scope of basic value ± 0.5mmol/L, promptly beginning infusion concentration is 20% glucose, adjusting glucose infusion speed (is glucose infusion rate, GIR), blood glucose value is controlled at about the scope of basic value ± 0.5mmol/L,, can continues the infusion glucose if blood glucose value exceeds the scope of basic value ± 0.5mmol/L.Surveyed a blood glucose in still per 10 minutes behind the infusion glucose, and in the shortest time, adjust glucose infusion rate, blood sugar recovery is arrived in the scope of basic value ± 0.5mmol/L according to blood glucose value.Carry out so repeatedly,, promptly reach steady statue when continuous three blood glucose values all are stabilized in the above-mentioned scope.(unit is mgkg with three GIR values that are in stable state of clamp ^-1Min ^-1) average.This index is used to judge whether to exist the degree of IR and IR.

1.3.6 the collection of pancreas in rat tissue and processing

After the off-test of the positive glucose clamp of hyperinsulinism, promptly put to death rat, open the abdominal cavity, between spleen, stomach and the free pancreas in duodenum crook, weigh behind the filter paper wipe dry, cut, get the tail of pancreas portion of tissue and place on the ice pan, be cut into 1mm rapidly along the pancreas longitudinal axis ³Fritter, 4 ℃ of glutaraldehydes are fixed, and are used for ultramicroscope and detect.

It is liquid-solid fixed that the fresh pancreatic tissue of part is put into Bouins, is used for morphologic detection; The fresh pancreatic tissue of part places rapidly in the frozen pipe, and liquid nitrogen is preserved, and is used for molecular Biological Detection.

1.3.7 the collection of rat liver tissue and processing

Part fresh liver tissue is put into 10% neutral formalin solution and is fixed, and is used for morphologic detection; The part fresh liver is organized and is placed rapidly in the frozen pipe, and liquid nitrogen is preserved, and is used for molecular Biological Detection.

1.3.8 the collection and the processing of fat in the rat abdomen

Put to death rat, get epididymis, mesentery, fold back peritoneum fat and weigh, count interior fat weight, and calculate interior fat weight percentage of liveweight percentage ratio; The all fat of the fresh testis of part is put into 10% neutral formalin solution and is fixed, and is used for morphologic detection.

1.3.9 the collection of rat skeletal muscle and processing

The fresh gastrocnemius tissue of part is put into 10% neutral formalin solution and is fixed, and is used for morphologic detection; The fresh gastrocnemius of part is organized and is placed rapidly in the frozen pipe, and liquid nitrogen is preserved, and is used for molecular Biological Detection.

1.3.10 rat skeletal muscle and liver AMPK detect early stage and handle

Take out liver of laboratory animal, muscular tissue respectively, and the 500 milligrams of tissue weights of weighing

Put 15 milliliters of taper centrifuge tubes of pre-cooling into

Adding GENMED cleaning liquid (Reagent A) cleans

Take out and clear up liquid

Be moved into a liquid nitrogen cryopreservation pipe

At once putting liquid nitrogen container into spends the night

Take out next day in liquid nitrogen container, at once with grinding rod pulverize be organized into Powdered

Put 15 milliliters of taper centrifuge tubes into

Add the GENMED lysate (Reagent B) that places in the ice groove

Powerful vortex concussion 30 seconds, fully mixing

Put the ice groove into and hatched 30 minutes, during powerful vortex concussion in per 10 minutes 30 seconds

At once put centrifugal 10 minutes of 4 ℃ of desk centrifuges into

Carefully pipette supernatant to new 1.5 milliliters of aseptic centrifuge tubes ,-80 ℃ of preservations

1.3.11 pancreas and liver HE staining procedure:

Paraffin is cut and is dewaxed to water.

Distilled water flushing 5 minutes.

Section was gone into the Harris haematoxylin 15 minutes.

Flowing water flushing 5 minutes.

0.5% hydrochloride alcohol differentiation 30 seconds

Flowing water flushing 10 minutes.

Yihong dyeing 10 minutes.Change tap water twice.

Gradient alcohol dehydration, the transparent mounting of dimethylbenzene I, II

1.3.12 statistical method

Adopt the analysis that takes statistics of SPSS16.0 software, the experimental data measurement data is with mean ± standard deviation

Expression, mean is relatively added up with the t check between two groups of Normal Distribution, relatively adopts one factor analysis of variance (One-wayANOVA) between group more than same batch, relatively adopts least significant difference (LSD) in twos.

2. result

2.1 laboratory animal ordinary circumstance

The OLETF rat is than the LETO obesity, and movable slow, lazyness is moved, hair color is withered matt.

Experimental session is got 5 of blood unexpected death animals, and wherein OLETF is 3,2 of LETO.

2.2 the rat food ration relatively

In experimental period, model group, dissipating depression of QI heat clearing away group, rosiglitazone group food ration be no significant difference relatively, and three groups of food rations all are higher than LETO group (P＜0.05).

Table 1 experimental rat food ration relatively

Annotate: with comparing age in week, model group and normal group are relatively ^##P＜0.01, ^#P＜0.05; Treatment group and model group are relatively ^*P＜0.01, ^*P＜0.05

2.3 the comparison of rat body constitution amount

As shown in table 2, in experimental period, model group, dissipating depression of QI heat clearing away group and rosiglitazone group body constitution amount all are significantly higher than LETO group (p＜0.01), and dissipating depression of QI heat clearing away group body constitution amount is starkly lower than model group and rosiglitazone group (p＜0.05)

The variation of table 2 experimental rat body constitution amount

2.4 the comparison of rat abdominal cavity fat content

Cuing open in 42 weeks extremely, the result shows, fat content obviously increases (P＜0.05) than the LETO group in the model group abdomen, dissipating depression of QI heat clearing away group reduces (P＜0.05) than model group, the rosiglitazone group increases (P＜0.05) than model group, see Table 3, the variation of this body weight and abdomen fat content may recover with islet function, and blood sugar level descends relevant.

The comparison of fat content, fat body ratio in the table 342W experimental rat abdomen

2.5 the variation of rat oral glucose tolerance test glucose level

As shown in table 4, model group and the AUC of LETO group before and after treatment be obvious difference (P＜0.01) all the time.With course of disease progress, model group AUC rises gradually, and two treatment groups and model group relatively AUC descend, when experiment finishes, dissipating depression of QI heat clearing away group and rosiglitazone group AUC all are starkly lower than model group (P＜0.01), and the rosiglitazone group is lower than dissipating depression of QI heat clearing away group slightly, but this species diversity does not have statistical significance.

Table 442W respectively organizes each time point blood glucose (mmol/L) of OGTT and AUG (mmolmin/L) changes

Annotate: AUC, with comparing age in week, compare ##P＜0.01, #P＜0.05 with normal group; Compare * * P＜0.01, * P＜0.05 with model group; Dissipating depression of QI heat clearing away group and rosiglitazone group are relatively ^{△ △}P＜0.01, ^△P＜0.05

2.6 the variation of experimental rat insulin secretion function

The tolerance test of rat oral glucose is with the variation of pacing insulin secretion and area under curve, the synchronous insulin area under curve of model group OGTT is organized greater than LETO, in conjunction with positive sugared clamp experimental result, the hints model group exists than obvious insulin resistance, after dissipating depression of QI heat clearing Chinese medicine and rosiglitazone treatment, two groups of insulin area under curve significantly increase, and point out two groups the B cell insulin secretion function had a better role.As shown in table 5

Table 542W respectively organizes each time point insulin (ng/ml) of OGTT and AUC (ngmin/ml) changes

Annotate: AUC, with comparing age in week, compare ##P＜0.01, #P＜0.05 with normal group; Compare * * P＜0.01, * P＜0.05 with model group

Pass through the synchronous insulin release test of 0GTT during 42W, we have calculated insulin secretion index (Δ I30/ Δ G30), the display model group obviously descends than LETO as a result, and dissipating depression of QI heat clearing away and rosiglitazone group obviously raise than model group, and dissipating depression of QI heat clearing away group is better than rosiglitazone.As shown in table 6

Insulin secretion index relatively (ng/mmol) is respectively organized in table 642W experiment

Annotate: compare ##P＜0.01, #P＜0.05 with normal group; Compare * * P＜0.01, * P＜0.05 with model group;

2.7 experiment is respectively organized the positive glucose clamp of hyperinsulinism result of the test relatively

As shown in table 7, model group rat stable state glucose infusion rate significantly descends than the LETO group, (P＜0.01), show that rat model has obvious insulin resistance, glucose infusion rate is significantly increased (P＜0.01) behind Chinese drug-treated group and the Western medicine group therapeutic intervention, shows that the two all has the good effect that improves insulin resistant.

The positive glucose clamp of hyperinsulinism result of the test is respectively organized in table 742W experiment

Annotate: compare ##P＜0.01, #P＜0.05 with normal group; Compare * * P＜0.01, * P＜0.05 with model group

2.8 the Blood Lipid situation is respectively organized in experiment

The triglyceride levels of model group extremely significantly increases, cholesterol, free fatty acid levels obviously increase, similar to the document record, meet OLETF Blood Lipid characteristics, the dissipating depression of QI heat clearing Chinese medicine has the effect of tangible triglyceride reducing, cholesterol, free fatty also there are the decline of statistical significance, show to have and transfers the fat effect preferably.As shown in table 8

Table 842W experiment is respectively organized the blood fat situation relatively

3 brief summaries

3.1OLETF-meet the doctor trained in Western medicine diabetes diagnosis, embody the animal model of pathogenesis feature

Raise through 30 all left and right sides times, part of O LETF rat blood sugar obviously raises, the oral administration carbohydrate tolerance test, and part rat blood sugar peak value＞16.7mmol/L and load back 120min blood glucose＞11.1mmol/L reach the diabetes diagnosis standard, the diabetes model success.

3.2 sugared quick spirit loses weight, and reduces fat content in the abdomen

This laboratory observation is organized all obviously increases of athero in body weight, the abdomen to OLETF than LETO.Through the treatment of 12 weeks, sugared quick spirit alleviates OLETF body constitution amount, and reduces fat content in the abdomen, compares with rosiglitazone, embodies greater advantage.

3.3 sugared quick spirit effectively improves sugar, lipid metabolism level

The result of study of this experiment shows, the abnormal carbohydrate metabolism of sugar quick spirit can improvement type 2 diabetes mellitus rat model, its hypoglycemic mechanism preliminary analysis is two aspects: the one, and the improvement of insulin resistant, we judge with the goldstandard of insulin sensitivity evaluation, there is obvious insulin resistance in the model group rat, Chinese drug-treated group and Western medicine group insulin resistant all obviously improve, its glucose infusion rate even greater than the normal control group, Chinese drug-treated group is compared with model group has significant difference, and this is consistent with the research team previous investigation; The 2nd, the improvement of islet cells secretory function, the result shows that the insulin area under curve of rat model obviously descends, Chinese drug-treated group and Western medicine group obviously improve after treating.Simultaneously, Chinese drug-treated group significantly improves insulin secretion index (Δ I30/ Δ G30), improves the pattern of insulin secretion.

The quick spirit of sugar also demonstrates transfers the fat effect preferably, has improved lipid metabolism in fat-reducing.Type 2 diabetes mellitus sugar, disorders of lipid metabolism are an integral body, the two connects each other, influence each other, therefore in treatment, to consider comprehensively, avoid independent blood sugar lowering or blood fat reducing treatment from the organic conception of disease, affect the treatment, must be placed on status of equal importance to blood sugar lowering, accent fat, this is for the process of control of diabetes, and the generation development that prevents and treats complication has great importance.

The quick spirit of experiment disaccharide is to the influence of spontaneous type 2 diabetes mellitus rat liver AMPK

1. materials and methods

1.1 laboratory animal (with experiment one)

1.2 grouping and administration (with experiment one)

1.3. the collection of experimental rat liver specimens and HE dyeing (with experiment one)

1.4. index and detection

1.4.1 liver AMPK active level is measured

Key instrument: Herolab mini desktop centrifuge, V-1800PC spectrophotometer and cuvette

Main agents: the AMPK of GENMED company organization kinase activity colorimetric assay detection kit

1.4.1.1 handle early stage

Take out laboratory animal fresh liver tissue respectively, and 500 milligrams of tissue weights of weighing

Put 15 milliliters of taper centrifuge tubes of pre-cooling into

Adding GENMED cleaning liquid (Reagent A) cleans

Take out and clear up liquid

Be moved into a liquid nitrogen cryopreservation pipe

At once putting liquid nitrogen container into spends the night

Put 15 milliliters of taper centrifuge tubes into

Add the GENMED lysate (Reagent B) that places in the ice groove

Powerful vortex concussion 30 seconds, fully mixing

At once put centrifugal 10 minutes of 4 ℃ of desk centrifuges into, speed is 10000g

Preparation of determine

Be ready to testing sample (for example lysis suspension etc.), place in the ice groove

Configure spectrophotometer (temperature is 30 ℃): wavelength is 340nm, 1 minute at interval, and reading 6 times (totally 5 minutes), and zero setting

1.4.1.2 background blank determination

Pipette GENMED buffer (Reagent C) to new cuvette

Add GENMED enzymatic liquid (Reagent D)

Add GENMED negative fluid (Reagent G)

Putting 30 ℃ of incubators into left standstill 2 minutes

Spectrophotometer, zero setting

Take out cuvette

1.4.1.3 sample determination

Pipette GENMED buffer (Reagent C) to new cuvette

Add GENMED enzymatic liquid (Reagent D)

Add 5 microlitre testing samples, put 30 ℃ of incubators into and left standstill 2 minutes

Add GENMED reactant liquor (Reagent E)

Add GENMED substrate solution (Reagent F)

Topple over for several times mixing (being limited within 3 seconds) up and down

At once put spectrophotometer into and detect, this is the sample reading:

0 minute-340 wavelength readings of 340 wavelength readings 5 minutes

According to formula, calculate AMPK enzymatic activity a quantitative index

1.4.2 the real-time fluorescence quantitative PCR method detects the mRNA of liver SREBP-1 and expresses

Key instrument

GeneAmp 5700TaqMAN PCR instrument U.S. Applied Biosystems company

Gel images analyser ImageMaster VDS U.S. pharmacia Biotech company

The High speed refrigerated centrifuge Germany BACKMAN Allagra.21R Centrifcige of company

The ultraviolet spectrophotometer Germany BACKMAN DU640 of company type

The OYY-III-5 of electrophresis apparatus Liuyi Instruments Plant, Beijing type

Main agents:

Trizol(Invitrogen)

RevertAidTM?First?Strand?cDNA?Synthesis?Kit(Fermentas)

SYBR?Green?PCR?Master?Mix(Applied?Biosystems)

Primer adopts primer premier5.0 design, and company is synthetic by the match Parkson.

1.4.2.1 from tissue, extract total RNA

● after getting about 100mg tissue and adding 1ml Trizol, be ground to that tissue is broken fully, the liquid thickness, room temperature placement 5 minutes makes its abundant cracking.

● press 200ul chloroform/mlTrizol and add chloroform, shake up 15 seconds, room temperature was placed 15 minutes.

● centrifugal 15 minutes of 4 ℃ of 12000g.

● draw the upper strata water, to the centrifuge tube of another pre-cooling.

● press 0.5ml isopropyl alcohol/ml Trizol and add isopropyl alcohol (pre-cooling) mixing, room temperature is placed 5-10min.

● 4 ℃ of centrifugal 10min of 12000g, abandon supernatant, RNA is sunken to the pipe end.

● add 75% ethanol (pre-cooling) by 1ml 75% ethanol/ml Trizol, gentle concussion centrifuge tube, precipitation suspends.

● 4 ℃ of centrifugal 5min of 7500g abandon supernatant as far as possible.

● room temperature is dried 5-10min.

● with 50 μ l RNase-free H ₂O, dissolving RNA sample, 55-60 ℃ of 10min.

● survey the quantitative RNA concentration of OD value.

1.4.2.2 the concentration of spectrophotometry nucleic acid

The spectrophotometer of band argon lamp used preceding pre-thermally-stabilised 15 minutes.

● draw 6 μ l RNA samples, add water to the 1.5ml mixing after, change in the spectrophotometric quartz cuvette.

● spectrophotometer is earlier with 1ml water suppressed zero.

● read optical density respectively at 260nm and 280nm, the concentration of RNA sample is: OD260* nucleic acid extension rate * 40/1000; Concentration unit is μ g/ μ l.

1.4.2.3RNA sample reverse transcription

● take out RNA template, reagent, after room temperature is melted, be placed on immediately in the ice bath, mixing on the DL device, of short duration centrifugal.

● the template degeneration:

Template?RAN x?ul(5.0ug)

Oligo(dT) ₁₈primer?0.5ug/ul 1.0ul

The DEPC treated water is to 12.0ul

Mixing, of short duration centrifugal, 70 ℃ of temperature were bathed 10 minutes, were placed in the ice bath at least 1 minute immediately.

● preparation reaction mixture (every pipe is by following amount preparation)

5×reaction?buffer 4.0ul

10mM?dNTP?Mix 2.0ul

RNase Inhibitor(20U/ul) 1.0ul

The mixed liquor amount of preparation is sample number N+2.

● every pipe adds the above-mentioned mixed liquor mixing of 7.0uL.37 ℃ of temperature were bathed 5 minutes.

● every pipe adds 1.0uL M-MuLV Reverse Transciptase (200u/uL), and 42 ℃ of temperature were bathed 60 minutes, and 70 ℃ of temperature were bathed 10 minutes.-20 ℃ of preservations are standby.

1.4.2.4 fluorescent quantitation PGR detects

● primer and reagent

SYBR?Green?PCR?Master?Mix：ABI(Applied?Biosystems)，Catalog?No.43049155

Primer sequence:

β-actin-FP：5’-GAG?ACC?TTC?AAC?ACC?CCA?GCC-3’

β-actin-RP:5 '-AAT GTC ACG CAC GAT TTC CC-3 ' amplified fragments is 263bp.

SREBP: amplified fragments is 168bp

The upstream: 5 '-CGC TAC CGT TCATCTATC AAT G-3 '

The downstream: 5 '-ACTTCG CAG GGT CAG GTT CT-3`

● the primer dilution: primer is diluted by final concentration 10pmol/ul, and concrete grammar is: V (ul)=O.D. value * 33ug/10M (the required DEPC treating water of V=volume, M=primer molecule amount)

● the preparation of reaction mixture:

Take out 2 * Buffer, RNase-free water, cDNA are placed in the room temperature, melt, be placed on immediately in the ice bath, and mixing on the DL device, of short duration centrifugal.

The contained reagent of everyone portion mixed liquor (reaction system V=25ul) sees the following form

Component?Volnme

2×SYBR?Green?PCR?Master?Mix， 12.5ul

xxx-F?primer(10pmol/ul) 1ul

xxx-R?primer(10pmol/ul) 1ul

Nuclease-free?H ₂O 8.5ul

Total?Volume 23ul

After preparing mixed liquor, add the 23ul mixed liquor in each reaction tube.

● application of sample: each reaction tube adds 2.0ul cDNA template, mixing on the DL device, of short duration centrifugal (less than 5sec).

● carry out at GeneAmp 5700 quantitative real time PCR Instruments, reaction condition is as follows:

95.0℃：10min

72.0℃：50sec

72.0℃：5min

More than reaction set all with computer that the PCR instrument links to each other on carry out, each circulation computer writes down the fluorescence signal value in the reaction tube automatically, and describes curve.Reaction finishes to calculate quantitative value automatically by PE 5700 software analysis results.

Experimental result is with the Ct value ratio of internal control gene and the genes of interest mRNA relative expression quantity as genes of interest, (because Ct value and the actual expression relation of being inversely proportional to, for intuitively, we get inverse with the Ct value ratio of genes of interest and Beta-actin when analytical data, just the Ct value ratio of Beta-actin and target gene is as the relative expression quantity of its mRNA)

1.4.3 the SABC method detects liver SREBP-1 protein expression

Instrument: various conventional reagent and equipment are provided by central laboratory of clinical research institute of China-Japan Friendship Hospital.LOW-70 ℃ of ultra cold storage freezer ULTRA, Japanese SANYO company; Oriental cherry flat push type microtome, Japanese SAKURA PTERATOMECRM-440; Exhibition sheet device, Japanese SAKURA Slide warner PS-53; Rotary automatic dehydration machine, Japanese SAKURATissue-Tek Tissue processor; Tissue embedding machine, Japanese SAKURA Tissue-Tek TECTM5SMC1-042-SO

Reagent: anti-Mus one anti antibody of SREBP-1 rabbit, Santa Cruz Biotechnology company product; Two anti-are the EnVisionTM system; The DAB liquid (all conventional reagent are Denmark DAKO company product) that develops the color.

1.4.3.1 fixing, dehydration, transparent, waxdip, embedding

After flesh tissue is fixing, gradient alcohol dehydration (45 minutes → dimethylbenzene of 75% ethanol 1 hour → 80% ethanol, 1 hour → 90% ethanol 1 hour → 95% ethanol, 1 hour → 100% ethanol 1 hour → dimethylbenzene of 1 hour → 100% ethanol 45 minutes), last waxdip embedding becomes wax stone.

1.4.3.2 slicing treatment

Microscope slide is handled: newly purchases microscope slide and soaked 24-48 hour through washing, and flowing water flushing 2-3 hour, distilled water immersion 1 hour, 60 ℃ of baking boxs are dried, and are soaked in the chromic acid gelatin solution of new preparation, pull out after soaking, and natural drying is standby.

On paraffin slicing machine, wax stone is cut into 4 microns slabs, tissue moved in 40 ℃ the constant-temperature hot water container, treat that tissue is launched fully after, invest on the microscope slide, lie on the exhibition sheet device, 56 ℃ of calorstats of the section of mounting spend the night, and take out standby.

1.4.3.3 immunohistochemical staining step:

Dewaxing, the aquation tissue slice.

The distilled water rinsing,

Drip 0.3%H ₂O ₂The blocking-up endogenous peroxydase was hatched 15 minutes.

The distilled water rinsing places TBS 5 minutes totally 3 times.

One anti-hatching 2 hours

The distilled water rinsing places TBS 5 minutes totally 3 times

Dripping EnvisionTM hatched 60 minutes.

TBS rinsing 5 minutes totally 3 times.

Color source DAB solution colour developing 3 minutes, the distilled water color development stopping.

Gradient alcohol dehydration, mounting glue mounting.

The microscopically observed result.

Establish negative control during experiment simultaneously, adopt TBS to replace one to resist as blank negative control.It is positive brown yellow granule to occur in the hepatocyte endochylema.

1.4.3.4 graphical analysis

The dyeing of the SREBP of liver and quantitative: adopt the medical image analysis systematic analysis of China Academy of TCM molecular immune chamber, 5 visuals field are selected in every section, every group is selected 8 sections at least, the gross area of measuring stained positive zone in each visual field is represented the quantity of this material cell, measure average optical (MOD) value and represent this expression of Substance amount of unit are cell, last average with every group compares.

1.4.4 statistical procedures

Finish measurement data mean ± standard deviation by the SPSS16.0 statistical package

Expression is relatively adopted one factor analysis of variance (One-wayANOVA) between group more than same batch, relatively adopts least significant difference (LSD) in twos.

2 results

2.1 the lipometabolic comparison of experimental rat (seeing experiment one)

2.2 liver lipid dystopy deposition conditions is respectively organized in experiment

Normal control group (LETO): portal area small artery, venule and gallbladder tube structure are normal, and hepatocyte is monokaryon or double-core, and the endochylema powder dyes, the even particle size unanimity.

Model group: liver rope arrangement disorder, no radial arrangement, the fat that differs in size in a large number in the endochylema drips; Some liver cell nuclears then by fat drop squeeze obvious off normal.

The rosiglitazone group: light microscopic fat down becomes, and swelling of liver cell is not obvious, has tiny fat to drip in the part cell.

Dissipating depression of QI heat clearing away group of the present invention: hepatocyte fat becomes not obvious, lobules of liver and hepatic sinusoid clear in structure, and cellularity is more complete.

2.3, the variation of experimental rat liver AMPK active level

Through the GENMED kit measurement, to compare with the LETO rat, model group AMPK enzymatic activity obviously reduces, and dissipating depression of QI heat clearing away group and rosiglitazone significantly rise than model group.As shown in table 9

Table 9 is respectively organized rat liver AMPK active level relatively

The present invention experimental results show that medicine sugar is quick and well-informedly overregulates liver AMPK enzymatic activity by above-mentioned, thereby improves type 2 diabetes mellitus people lipid metabolism level, reduces the deposition of lipid in peripheral tissues, reaches the effect of treatment and prevention type 2 diabetes mellitus.

In addition, the quick well-informed hypothalamic AMPK enzyme that overregulates of sugar, thereby control trophic behavior.

The specific embodiment

The invention will be further described by following specific embodiment, but not as limitation of the present invention.

The preparation of embodiment 1, the quick curing capsule of sugar of the present invention

Get Radix Trichosanthis 30g, Radix Bupleuri 12g, Fructus Aurantii Immaturus 9g, Radix Et Rhizoma Rhei 3g, Rhizoma Pinelliae 6g, Radix Scutellariae 9g, Rhizoma Coptidis 6g, Radix Paeoniae Alba 9g, Fructus Mume 15g, Fructus Crataegi 9g, Rhizoma Polygonati 9g, Radix Ginseng 9g, Fructus Momordicae charantiae 9g;

Add 4 times of amount ethanol of 80%, 60 ℃ of reflux, extract, 3 times, each 1.5 hours, merge extractive liquid,, ethanol are reclaimed in 70 ℃ of pressurizations, 80 ℃ of vacuum dryings were pulverized 80 mesh sieves, drug extract;

Add lactose and make capsule.

The preparation of embodiment 2, the quick clever tablet of sugar of the present invention

Add dextrin and make tablet.

The preparation of embodiment 3, the quick clever granule of sugar of the present invention

60 ℃ of forced air dryings are granulated, and granulate promptly gets granule.

The preparation of embodiment 4, the quick drop pills of sugar of the present invention

The Polyethylene Glycol that adds, mix homogeneously, fusion, last drop pill machine is made drop pill.

The preparation of embodiment 5, the quick clever oral cavity disintegration tablet of sugar of the present invention

Add 5% polyvinylpolypyrrolidone, 0.1% magnesium stearate, 50% microcrystalline Cellulose is made soft material with an amount of alcoholic solution, granulates, and 60 ℃ of forced air dryings are granulated, granulate, compacting promptly gets oral cavity disintegration tablet in flakes.

The preparation of embodiment 6, the quick curing capsule agent of sugar of the present invention

Add appropriate amount of starch, sucrose and magnesium stearate are granulated, and incapsulate, and promptly get capsule.

The preparation of embodiment 7, the quick clever tablet of sugar of the present invention

With starch, sodium carboxymethyl cellulose, Pulvis Talci mix homogeneously are granulated, and tabletting promptly gets tablet.

The preparation of embodiment 8, the quick Ling oral liquid of sugar of the present invention

With syrup 4g, be dissolved in the pure water of 100ml, homogenizing filters, through high-temperature short-time sterilization (135 ℃, 4s).Sterile filling, packing make oral liquid.

The preparation of embodiment 9, the quick Ling oral liquid of sugar of the present invention

With syrup 5g, be dissolved in the pure water of 100ml, homogenizing filters, through high-temperature short-time sterilization (135 ℃, 4s).Sterile filling, packing make oral liquid.

The preparation of embodiment 10, the quick curing capsule of sugar of the present invention

Get Radix Trichosanthis 9g, Radix Bupleuri 12g, Fructus Aurantii Immaturus 9g, Radix Et Rhizoma Rhei 3g, Rhizoma Pinelliae 6g, Radix Scutellariae 9g, Rhizoma Coptidis 6g, Radix Paeoniae Alba 9g, Fructus Mume 9g;

Add lactose and make capsule.

The preparation of embodiment 11, the quick curing capsule of sugar of the present invention

Get Radix Trichosanthis 30g, Radix Bupleuri 12g, Fructus Aurantii Immaturus 9g, Radix Et Rhizoma Rhei 3g, Rhizoma Pinelliae 6g, Radix Scutellariae 9g, Rhizoma Coptidis 6g, Radix Paeoniae Alba 9g, Fructus Mume 15g, Fructus Crataegi 9g;

Add lactose and make capsule.

The preparation of embodiment 12, the quick clever tablet of sugar of the present invention

Add dextrin and make tablet.

The preparation of embodiment 13, the quick clever tablet of sugar of the present invention

Add dextrin and make tablet.

The preparation of embodiment 14, the quick clever granule of sugar of the present invention

The preparation of embodiment 15, the quick clever granule of sugar of the present invention

The preparation of embodiment 16, the quick drop pills of sugar of the present invention

The preparation of embodiment 17, the quick drop pills of sugar of the present invention

The preparation of embodiment 18, the quick clever oral cavity disintegration tablet of sugar of the present invention

The preparation of embodiment 19, the quick clever oral cavity disintegration tablet of sugar of the present invention

The preparation of embodiment 20, the quick curing capsule agent of sugar of the present invention

The preparation of embodiment 21, the quick curing capsule agent of sugar of the present invention

The preparation of embodiment 22, the quick curing capsule agent of sugar of the present invention

The preparation of embodiment 23, the quick curing capsule agent of sugar of the present invention

The preparation of embodiment 24, the quick curing capsule agent of sugar of the present invention

The preparation of embodiment 25, the quick curing capsule agent of sugar of the present invention

The preparation of embodiment 26, the quick clever injectable powder of sugar of the present invention

Add glucose 4.5g, sodium thiosulfate 0.9g and distilled water 1ml again, behind the said components mix homogeneously, lyophilization, packing promptly gets injectable powder.

The preparation of embodiment 27, the quick clever injectable powder of sugar of the present invention

The preparation of embodiment 28, the quick clever injectable powder of sugar of the present invention

Group component in the foregoing description can enlarge or reduce in scale simultaneously according to producing needs.

Claims

1. the application of pharmaceutical composition in the medicine of preparation raising AMPK enzymatic activity, wherein said pharmaceutical composition is that prescription is formulated by following parts by weight of Chinese traditional medicine crude drug: Radix Trichosanthis 5-40 part, Radix Bupleuri 10-30 part, Fructus Aurantii Immaturus 3-15 part, Radix Et Rhizoma Rhei 1-6 part, Rhizoma Pinelliae 1-12 part, Radix Scutellariae 3-15 part, Rhizoma Coptidis 1-12 part, Radix Paeoniae Alba 3-15 part, Fructus Mume 5-20 part.

2. application according to claim 1, it is characterized in that, Fructus Crataegi in can also adding in the pharmaceutical composition prescription, the prescription that obtains is: Radix Trichosanthis 10-30 part, Radix Bupleuri 10-30 part, Fructus Aurantii Immaturus 3-15 part, Radix Et Rhizoma Rhei 1-6 part, Rhizoma Pinelliae 1-12 part, Radix Scutellariae 3-15 part, Rhizoma Coptidis 1-12 part, Radix Paeoniae Alba 3-15 part, Fructus Mume 5-20 part, Fructus Crataegi 3-15 part.

3. application according to claim 1, it is characterized in that, Fructus Crataegi in can also adding in the pharmaceutical composition prescription, Rhizoma Polygonati, Radix Ginseng and Fructus Momordicae charantiae, the prescription that obtains is: Radix Trichosanthis 10-30 part, Radix Bupleuri 10-30 part, Fructus Aurantii Immaturus 3-15 part, Radix Et Rhizoma Rhei, 1-6 part, Rhizoma Pinelliae 1-12 part, Radix Scutellariae 3-15 part, Rhizoma Coptidis 1-12 part, Radix Paeoniae Alba 3-15 part, Fructus Mume 5-20 part, Fructus Crataegi 3-15 part, Rhizoma Polygonati 3-15 part, Radix Ginseng 3-15 part, Fructus Momordicae charantiae 3-15 part.

4. application according to claim 1 is characterized in that, the pharmaceutical composition prescription is: 9 parts of Radix Trichosanthis, 12 parts of Radix Bupleuri, 9 parts of Fructus Aurantii Immaturuss, 3 parts of Radix Et Rhizoma Rhei, 6 parts of the Rhizoma Pinelliaes, 9 parts of Radix Scutellariaes, 6 parts of Rhizoma Coptidis, 9 parts of the Radix Paeoniae Albas, 9 parts of Fructus Mumes.

5. application according to claim 2 is characterized in that, the pharmaceutical composition prescription is: 30 parts of Radix Trichosanthis, 12 parts of Radix Bupleuri, 9 parts of Fructus Aurantii Immaturuss, 3 parts of Radix Et Rhizoma Rhei, 6 parts of the Rhizoma Pinelliaes, 9 parts of Radix Scutellariaes, 6 parts of Rhizoma Coptidis, 9 parts of the Radix Paeoniae Albas, 15 parts of Fructus Mumes, 9 parts of Fructus Crataegis.

6. application according to claim 3, it is characterized in that, the pharmaceutical composition prescription is: 30 parts of Radix Trichosanthis, 12 parts of Radix Bupleuri, 9 parts of Fructus Aurantii Immaturuss, 3 parts of Radix Et Rhizoma Rhei, 6 parts of the Rhizoma Pinelliaes, 9 parts of Radix Scutellariaes, 6 parts of Rhizoma Coptidis, 9 parts of the Radix Paeoniae Albas, 15 parts of Fructus Mumes, 9 parts of Fructus Crataegis, 9 parts of Rhizoma Polygonatis, 9 parts of Radix Ginsengs, 9 parts in Fructus Momordicae charantiae.

7. according to any described application of claim 1-3, it is characterized in that described pharmaceutical composition passes through to regulate liver AMPK enzymatic activity, thereby improves type 2 diabetes mellitus people lipid metabolism level.

8. according to any described application of claim 1-3, it is characterized in that described pharmaceutical composition is by adjusting liver AMPK enzymatic activity, thereby the reduction lipid is in the deposition of peripheral tissues.

9. according to any described application of claim 1-3, it is characterized in that described pharmaceutical composition is by the hypothalamic AMPK enzymatic activity of adjusting, thus the control trophic behavior.

10. according to any described application of claim 1-3, it is characterized in that described pharmaceutical composition is by regulating the AMPK enzymatic activity, treatment and prevention type 2 diabetes mellitus.