CN103432207A - Preparation method of fructus forsythiae extract lignan and application of extract in blood sugar reducing drugs - Google Patents
Preparation method of fructus forsythiae extract lignan and application of extract in blood sugar reducing drugs Download PDFInfo
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Abstract
The invention relates to a preparation method of a fructus forsythiae extract lignan and an application of the extract in blood sugar reducing drugs. The problem that a substance for preparing the blood sugar reducing drugs can be extracted from fructus forsythiae can be effectively solved. The extract is obtained through the following steps of crushing fructus forsythiae into medical powder, soaking the medical powder in ethanol for 0.5 h, carrying out reflux extraction twice, abandoning filter residue to obtain an extract liquid, performing decompression concentration on the extract liquid until no alcohol smell is left, adjusting the concentration of the medical liquid to be equivalent to contain 0.5 g/ml of a crude drug, thereby obtaining a sample liquid; applying the sample liquid to a type AB-8 macroporous adsorption resin column; eluting by using water in an amount which is 8 times of the volume of the column to obtain and abandon an eluate I; next, eluting by using ethanol in an amount which is 8 times of the volume of the column so as to obtain and abandon an eluate II; and finally, eluting by using ethanol in an amount which is 10 times of the volume of the column so as to obtain an eluate III, recovering the ethanol, and then carrying out decompression concentration and drying to obtain the fructus forsythiae lignan. According to the invention, the raw materials are rich, the preparation method is simple, the prepared fructus forsythiae lignan has the effect of reducing blood sugar and can be effectively used for preparing the blood sugar reducing drugs; therefore, a new application of treating diabetes is developed.
Description
Technical field
The present invention relates to medicine, particularly the preparation method of a kind of Fructus Forsythiae extract lignanoid and the application in hypoglycemic medicine thereof.
Background technology
Diabetes be by environmental factors and inherited genetic factors interact and cause take the chronic metabolic disease that chronic hyperglycemia is common trait, domestic to its symptom so-called " hyperglycemia ".Long-term hyperglycemia can cause microvascular endothelial to produce pathological changes, and the protein of following, lipid metabolism can cause the abnormal of the organizational structure such as the heart, brain, kidney, nerve, blood vessel and function extremely, even can cause the organ failure and threat to life.Fruity gives its another name " reticent killer ", the particularly diabetes of adult type, and diabetics on average can reduce by 10 year life-span.And the complication occurred can spread all over whole body, and the complication of diabetes comprises: self-disciplining nervous disorder, neurological disorder, cerebral thrombosis, cerebral infarction.Cataract, decayed tooth, stomatitis, bronchitis, dermatosis, myocardial infarction, uremia, pedopathy change etc.Asia diabetes association reveals, by 2012, the total number of persons of middle national trouble diabetes reached 9,240 ten thousand, and had following characteristics: 1 sickness rate is high, can reach 9.7%; 2 rejuvenation trend, the between twenty and fifty shortage, move, and causes diabetes occurred frequently; 3 exist the treatment mistaken ideas, and the time of diabetes mellitus in China patient infusion insulin is generally more late.China is populous nation, the population proportion is large, Chinese are also the crowds that sends out well of diabetes simultaneously, studies show that, at rich countries, Chinese's prevalence is many more than 10%, and the sickness rate rising of nearly 20 years China's diabetes rapidly, caused the Chinese government, hygiene department and numerous medical workers' attention.
Diabetes are the metabolic diseases that caused by different pathogeny, and feature is chronic hyperglycemia state and carbohydrate, fat and the protein metabolism disorder caused due to defect of insulin secretion or insulin action defect.Its chronic complicating diseases has nephropathy, neuropathy etc., because of the oral antidiabetic convenience, fast, no pain, the advantages such as sustainable prolonged application, clinical practice is very extensive, treatment diabetes at present commonly used and the medicine of complication have several classes: 1 biguanides, recover the inhibitory action of insulin to adenylate cyclase activity by liver plasma membrane G albumen, allophone and the hepatic glucose output of hepatic glycogen have been reduced, promote the anerobic glycolysis of myocyte's glucose, the picked-up of the peripheral tissues such as increase muscle to glucose, strengthen the sensitivity of tissue to insulin, to the normal person without blood sugar reducing function.Represent that medicine has metformin etc., metformin can increase lactic acid gathering in blood, can cause lactic acidosis and with the gastrointestinal untoward reaction.2 sulphanylureas, Main Function is in the pancreatic beta cell uelralante.Represent that medicine has glibenclamide, gliclazide, gliquidone etc.3 glucose glycoside enzyme inhibitors, represent that medicine has acarbose, miglitol etc., competitive inhibition glucosidase in gastrointestinal tract, delay disaccharidase, oligosaccharide and polysaccharide in enteric cavity and discharge glucose, slight appetite-suppressing, finally reduce post-prandial glycemia, and do not cause hypoglycemia.4 euglycemic agents, represent that medicine has rosiglitazone, pioglitazone etc., is can improve skeletal muscle and adipose cell to the absorption of glucose and reduce the output of glycogen, the sensitivity of raising to insulin, improve insulin-resistant states, improve carbohydrate metabolism, reduce blood glucose value.5 non-sulphanylureas class Insulin-secreting agents, represent that medicine has repaglinide, Nateglinide etc., and on the beta Cell of islet film, specific receptor is combined, and makes the pancreatic beta cell depolarization, thereby promote insulin secretion.6 dipeptidyl peptidase-4 inhibitors, represent that medicine has sitagliptin, and the activity of DPP-4, can protect incretin not to be degraded always, stimulating pancreas β cell regeneration.
But western medicine has side effect more, can bring Liver and kidney toxicity, along with the prevalence of diabetics constantly rises, the side effect that Western medicine in the past brings and insulin use procedure China are difficult to control, make a lot of experts and scholars start to find the new drug for the treatment of diabetes, from Chinese medicine, the screening effective ingredient is a new effective way, becomes domestic and international expert's study hotspot.
The traditional Chinese medical science thinks that diabetes spp is in the category of quenching one's thirst, and in "Nei Jing", the cause of disease, pathogenesis, the card of primary disease has been controlled to clear and definite discussion.Diabetes clinical symptoms performance is complicated and changeable, is due to due to a plurality of visceral dysfunctions, the closest with the relation of spleen, kidney.The modern study discovery, the genesis of blood stasis and diabetes has important impact, and blood stasis is through diabetes all the time.In diabetic duration, cause the reason of blood stasis roughly as follows: 1 deficiency of YIN causes the stasis of blood, and the deficiency of YIN is one of diabetes the important pathogenesis, YIN fluid being insufficient, and due to homogeny of clear fluid and blood, body fluid deficiency is insufficiency of blood, causes blood circulation not smooth, and the blood vessels operation does not freely produce blood stasis.The 2 scorching stasis of bloods that cause, diabetics is because cloudy body fluid deficiency consumes, and insufficiency of water leading to hyperactivity of fire has scorching flourishingly more, decocts cloudy blood, makes blood concentrated, and sering is jammed and is produced blood stasis.3 deficiency of vital energys cause the stasis of blood, and diabetics is scorching interior vehement, and not only impairment of body fluid also can consume gas, causes deficiency of both QI and YIN, and the deficiency of vital energy promotes unable, and blood is slow and produce blood stasis.4 stagnation of QIs cause the stasis of blood, and feelings will does not freely the most easily cause depression and stagnation of QI, and the stagnation of QI affects the blood vessels operation and produces blood stasis.5 prolonged illness must the stasis of blood, and diabetes are as a kind of chronic lifelong participation disease, and its course of disease delay, with the passing of time do not heal, and the traditional Chinese medical science is thought pathogen usually intruding into collateral in protracted disease, and the channels stasis of blood is stagnant, hematogenous blockage and produce blood stasis.Once blood stasis forms, and can further cause the development of diabetes and increase the weight of as new paathogenic factor again.And pyretic toxicity plays an important role in the carcinogenesis of human of diabetic duration, diabetics all is accompanied by scorching clinically, body constitution is biased, worked with one's mind excessively, and depressed emotion, labor stringency, and outer temperature-sensing thermal poison is evil, all can cause burning hot yin-damaging and Qi consuming, and intimately flourishing, or pent up with the passing of time, can turn to pyretic toxicity." blood circulation promoting and blood stasis dispelling, the heat-clearing and toxic substances removing " blood sugar lowering of should take is the Therapeutic Principle.
The dry fruit that Fructus Forsythiae is Oleaceae forsythia Fructus Forsythiae, for Chinese medicine, being gone through an edition Pharmacopoeia of the People's Republic of China records, bitter in the mouth, row is slightly cold, and enters the heart, lung, small intestine meridian, has the effect of heat-clearing and toxic substances removing, mass dissipating and swelling eliminating, dispelling wind and heat pathogens, clinical be used for the treatment of calentura from the beginning of, anemopyretic cold, heating, vexed, laryngopharynx swelling and pain, macule, erysipelas etc., have antibiotic, antiviral, antiinflammatory, antiendotoxin, protect the liver, diuresis, analgesic, antiallergic, the effects such as blood sugar lowering, but less for the treatment report of diabetes clinically.Fructus Forsythiae is rich in plurality of active ingredients, as compositions such as flavone, saponin, lignanoids, polysaccharide, wherein the Fructus Forsythiae ester glycoside in lignanoid is one of the antioxidation of finding from forsythia up to now, composition that antibacterial activity is the strongest, and is the principal character composition of Fructus Forsythiae.So how it is extracted, realize the application of Fructus Forsythiae extract in preparing hypoglycemic medicine, be that people expect the technical problem solved.
Summary of the invention
For above-mentioned situation, for overcoming the defect of prior art, the present invention's purpose just is to provide the preparation method of a kind of Fructus Forsythiae extract lignanoid and the application in hypoglycemic medicine thereof, can effectively solve from Fructus Forsythiae and extract the material for the preparation of hypoglycemic medicine.
The technical scheme that the present invention solves is, the application of Fructus Forsythiae extract in preparing hypoglycemic medicine, this extract is that Fructus Forsythiae powder is broken into medicated powder, medicated powder soak with ethanol 0.5h, reflux, extract, 2 times, filtering residue discards, and obtains extracting solution, and extracting solution is evaporated to without the alcohol flavor, adjust liquor strength and containing crude drug 0.5g/ml, obtain sample liquid for being equivalent to; AB-8 type macroporous adsorptive resins on sample liquid, first use the water elution of 8 times of column volumes, obtains the eluent I and discard; With the ethanol elution of 8 times of amount column volumes, obtain the eluent II and discard again; Finally, with the ethanol elution of 10 times of amount column volumes, obtain the eluent III, reclaim ethanol, the concentrating under reduced pressure drying, obtain Fructus Forsythiae lignanoid.
Abundant raw material of the present invention, preparation method is simple, and the Fructus Forsythiae lignanoid of preparation has blood sugar reducing function, is effective to prepare hypoglycemic medicine, and the new purposes of developing treatment diabetes, be the innovation on hypoglycemic medicine.
The specific embodiment
Below in conjunction with concrete condition, the specific embodiment of the present invention is elaborated.
The present invention is in concrete enforcement, by following technical scheme, provided: the application of Fructus Forsythiae extract in preparing hypoglycemic medicine, this extract is that Fructus Forsythiae powder was broken into the medicated powder of 60-80 mesh sieve, the soak with ethanol 0.5h that for medicated powder, 8 times of weight, volumetric concentration are 80%, reflux, extract, 2 times, the 1st 1.5h, the 2nd 1h, filtering residue discards, obtain extracting solution, extracting solution is evaporated to without the alcohol flavor, adjusts liquor strength and containing crude drug 0.5g/ml, obtains sample liquid for being equivalent to; AB-8 type macroporous adsorptive resins on sample liquid, the ratio of applied sample amount and resin volume is 1:12, the resin column blade diameter length ratio is 1:12, first uses the water elution of 8 times of column volumes, obtains the eluent I and discards; With 8 times of column volume, ethanol elutions that volumetric concentration is 10% of amount, obtain the eluent II and discard again; Finally, with 10 times of column volume, ethanol elutions that volumetric concentration is 75% of amount, obtain the eluent III, reclaim ethanol, the concentrating under reduced pressure drying, obtain Fructus Forsythiae lignanoid, content >=58%; This Fructus Forsythiae lignanoid is effective to prepare hypoglycemic medicine, realizes the application of Fructus Forsythiae lignanoid in preparing hypoglycemic medicine.
The inventive method is simple, and abundant raw materials is easy to prepare Fructus Forsythiae lignanoid, and this Fructus Forsythiae lignanoid has blood sugar reducing function, can be used for preparing hypoglycemic medicine, and has obtained satisfied effect through experiment, and relevant experimental data is as follows:
1 impact on epinephrine induced mice hyperglycemia model
1.1 pharmaceutical agent
Fructus Forsythiae of the present invention lignanoid;
Metformin hydrochloride tablet, Shanghai Sine Pharmaceutical Co., Ltd. produces, lot number 120523; Adrenalin hydrochloride, Tianjin gold credit aminoacid company limited is produced, lot number 20120308: normal saline, Zhengzhou Yonghe Pharmaceutical Co, lot number 12042401; Sodium carboxymethyl cellulose, good fortune chemical reagent factory in morning in Tianjin is produced, lot number 20090826; The glucose assays test kit, produced and provided by Zhongsheng Beikong Biological Science & Technology Co., Ltd., lot number 120491; Hepatic glycogen is measured test kit, builds up bio-engineering research by Nanjing and is produced, lot number 20120728.
1.2 experimental apparatus
FA (N)/JA (N) series electronic balance, Shanghai Min Qiao precision instrument company limited; The UV-2000 ultraviolet-uisible spectrophotometer, Shanghai Techcomp Instrument Ltd. produces; The TGL-16G desk centrifuge, Anting Scientific Instrument Factory, Shanghai produces; DZKW-4 type electronic thermostatic water-bath, in Beijing, emerging great achievement Instrument Ltd. produces; The KDC-160HR High speed refrigerated centrifuge, in Keda Innovation Co., Ltd, good branch company produces; The BIORAD-680 microplate reader, 680Microplate Reader, Bio-Rad Laboratories; Adjustable pipette, produced by Shanghai Lei Bo Analytical Instrument Co., Ltd.
1.3 laboratory animal
Kunming mouse, male, body weight 18～22g, Hebei province's Experimental Animal Center provides, the quality certification number: 1209018; Laboratory quality certification SYXK (Henan) 2010-001.
1.4 test method
Adopt abdominal cavity to annotate adrenergic method and set up the mice hyperglycemia model.Before modeling, metformin group, the large, medium and small dosage group of Fructus Forsythiae lignanoid give respectively corresponding medicine gavage, and blank group and model group give same volume 0.1%CMC gavage, every day 1 time, successive administration 7 days.In administration the 7th day (fasting 8h), after administration 1h, except normal group, other 11 groups of lumbar injection adrenalin hydrochlorides (0.2mg/kg), the normal group lumbar injection is with the normal saline of volume, and 0.5h posterior orbit venous blood collection, adopt the determination of glucose oxidase blood glucose value, cut off abdominal cavity, get same position hepatic tissue and measure hepatic glycogen content.
1.5 statistical method
Data analysis carries out the statistical procedures of data information, average ± standard deviation for measurement data with the medical statistics bag of SPSS 17.0 for windows
mean, relatively adopt one factor analysis of variance between each group, the neat person of variance test adopts the Ridit check with least significant difference (LSD) method, heterogeneity of variance person with Games-Howell method check, ranked data.
1.6 result
Table 1 on the impact of blood glucose value and the hepatic glycogen content of epinephrine induced mice hyperglycemia model (
n=12)
* with model group than P<0.01
From table can find out: with blank group ratio, model group blood glucose significantly raises, and hepatic glycogen content significantly reduces, and modeling success is described.With the model group ratio, metformin group, Fructus Forsythiae lignanoid each dosage group of each dosage group all can significantly reduce the blood glucose value (P<0.01) after the modeling mice; Hepatic glycogen content (P<0.01) can significantly raise.Illustrate that Fructus Forsythiae lignanoid, metformin all can be to the glycogenolytic phenomenons of the aggravation regulating liver-QI of the glyconeogenesis due to antiadrenergic drug.
2 impacts on rat diabetes model due to streptozotocin
2.1 pharmaceutical agent
Fructus Forsythiae lignanoid;
Metformin hydrochloride tablet, Hisense's friendship pharmaceutical factory company limited is produced, lot number 120523; Streptozotocin, sigma company produces, lot number Z120315; Normal saline, Zhengzhou Yonghe Pharmaceutical Co, lot number 12042401; Glacial acetic acid (analytical pure), permanent Xinghua, Tianjin is learned reagent Manufacturing Co., Ltd and is produced, lot number 20110103; Formaldehyde, Yantai City's chemical industry company limited in pairs produces, lot number 20111101; Blood sugar kit, Zhongsheng Beikong Biological Science & Technology Co., Ltd. produces, lot number 120511; The NOS test kit, Nanjing is built up bio-engineering research and is produced, lot number 20121208; The SOD test kit, Nanjing is built up bio-engineering research and is produced, lot number 20121215; Glycated serum protein (GSP) testing cassete, Nanjing is built up bio-engineering research and is produced, lot number 20121019; The nitrogen oxide testing cassete, Nanjing is built up bio-engineering research and is produced, lot number 20121212; The Triglyceride Reagent box, the safe clinical reagent company limited of Beijing Northization is produced, lot number 20121105; The T-CHOL test kit, the safe clinical reagent company limited of Beijing Northization is produced, lot number 20120808; RatIAA ELISA Kit test kit, R& D company produces, LOT:20121101A; Rat INS ELISA Kit test kit, R& D company produces, LOT:20121101A.
2.2 experimental apparatus
FA (N)/JA (N) series electronic balance, Shanghai Min Qiao precision instrument company limited; The UV-2000 ultraviolet-uisible spectrophotometer, Shanghai Techcomp Instrument Ltd. produces; The TGL-16G desk centrifuge, Anting Scientific Instrument Factory, Shanghai produces; DZKW-4 type electronic thermostatic water-bath, in Beijing, emerging great achievement Instrument Ltd. produces; The KDC-160HR High speed refrigerated centrifuge, in Keda Innovation Co., Ltd, good branch company produces; The BIORAD-680 microplate reader, 680Microplate Reader, Bio-Rad Laboratories; Adjustable pipette, produced by Shanghai Lei Bo Analytical Instrument Co., Ltd.
2.3 laboratory animal
Clean level SD rat, male, 180-220g, Hebei province's Experimental Animal Center provides, the animal quality certification number: 1210019, laboratory quality certification SYXK (Henan) 2010-001.
2.4 test method
Adopt the method for sublingual vein annotation streptozotocin to set up the rat diabetes model.Buy the normal raising of animal 3 days, according to body weight, the streptozotocin of sublingual vein injection 10mg/ml concentration, dosage is 40mg/kg, tail venous blood sampling after 5 days, side serum blood glucose value, and choose blood glucose value>16.0mg/L, and the rat that obviously there is polydipsia polyphagia polyuria symptom, according to the random evenly grouping of blood glucose value, metformin group, the large, medium and small dosage group of Fructus Forsythiae lignanoid give respectively the relative medicine gavage, blank group and model group give same volume 0.1%CMC gavage, every day 1 time, administration every day 1 time, successive administration 30 days.Respectively at the 10th, 20,30 days of administration, after when perfusion 2, tail vein blood, separation of serum, survey fasting 6 hours blood glucose values, after last administration 2 hours, eyeball is got blood, surveys serum insulin (Ins), insulin antibody, NO level and NOS vigor, triglyceride (TG), cholesterol (TC), superoxide dismutase (SOD), glycation serum protein.Put to death rat after getting blood, get pancreas, kidney is done section, by its pathological change of electron microscopic observation.
2.5 statistical method
Data analysis carries out the statistical procedures of data information, average ± standard deviation for measurement data with the medical statistics bag of SPSS 13.0 for windows
mean, relatively adopt one factor analysis of variance between each group, the neat person of variance test adopts the Ridit check with least significant difference (LSD) method, heterogeneity of variance person with Games-Howell method check, ranked data.
2.6 result
Table 2 on the impact of rat diabetes model SOD, NO, NOS due to streptozotocin (
n=12)
* with model group than P<0.01, * with model group than P<0.05
From table can find out: with blank group ratio, model group SOD vigor significantly reduces (P<0.01), NO content, NOS vigor significantly raise (P<0.01) illustrate that modeling treated animal oxidation resistance reduces, the NO metabolism disorder.With the model group ratio, metformin group (P<0.01) the SOD enzyme activity that can significantly raise, significantly reduce (P<0.01) NO content and NOS enzyme activity.The big or middle dosage group of Fructus Forsythiae lignanoid (P<0.01) the SOD enzyme activity that can raise significantly, significantly reduce (P<0.01) NO level and NOS enzyme activity, Fructus Forsythiae lignanoid small dose group (P<0.01) the SOD enzyme activity (P<0.01) that can significantly raise, obviously reduce (P<0.05) NOS enzyme activity.
Table 3 on the impact of rat diabetes model T-CHOL, triglyceride due to streptozotocin (
n=12)
* with model group than P<0.01, * with model group than P<0.05
As can be seen from the above table, with blank group, compare, model group T-CHOL, content of triglyceride significantly rise (P<0.01), illustrate that diabetes model can cause disorders of lipid metabolism.With model group, compare, the metformin group can reduce the content of (P<0.01) triglyceride and T-CHOL significantly, effectively the correcting lipid disorder phenomenon.The large, medium and small dosage group of Fructus Forsythiae lignanoid, the large, medium and small dosage group of Herba Scutellariae Barbatae flavone, the large, medium and small dosage group of Herba Violae flavone can significantly reduce the content of (P<0.01) T-CHOL, triglyceride, correcting lipid disorder phenomenon.
Table 4 on the impact of rat diabetes model glycated serum protein, insulin, insulin antibody due to chain urea assistant bacterium (
n=12)
* with model group than P<0.01, * with model group than P<0.05
As can be seen from the above table: compare model group can significantly raise glycated serum protein content and insulin antibody level (P<0.01) with blank group, can reduce significantly insulin content (P<0.01), the modeling success is described.Compare the metformin group with model group and can significantly reduce (P<0.01) glycated serum protein content, insulin antibody level, insulin content (P<0.01) can significantly raise.The large, medium and small dosage group of Fructus Forsythiae lignanoid can significantly reduce (P<0.01) glycated serum protein content, and (P<0.01) insulin level significantly raises; The big or middle dosage group of Fructus Forsythiae lignanoid can significantly reduce (P<0.05) insulin antibody level.
Table 5 on the impact of rat diabetes model dynamic glucose value due to streptozotocin (
n=12)
* with model group than P<0.01, * with model group than P<0.05
From table can find out: except the blank group, each organizes the equal no significant difference of blood glucose before administration, illustrate that grouping is even.With blank group ratio, at the 10th, 20,30 days, the equal significance of model group blood glucose raise (P<0.01), and the success of modeling type is described.With the model group ratio, administration the 10th day, each administration group was little to blood sugar influence, and administration the 20th day, the 30th day, the large, medium and small dosage group of Fructus Forsythiae lignanoid and metformin group all can significantly reduce blood sugar level (P<0.01).Illustrate that Fructus Forsythiae lignanoid can effectively reduce the animal pattern blood glucose value, improves diabetic symptom.
The impact of table 6 on rat diabetes model pancreatic tissue due to streptozotocin
In "-" pancreatic tissue, islets of langerhans is more than 4, and the islet cells endochylema is abundant, and nucleus is loose; In "+" pancreatic tissue, islets of langerhans is more than 4, the atrophy of minority islet cells endochylema, and a few cell core is intensive; In " ++ " pancreatic tissue, islets of langerhans is individual at 2-3, the atrophy of part islet cells endochylema, and the parts of fine karyon is intensive; In " +++" pancreatic tissue, islets of langerhans is at 0-1, and all the islet cells endochylemas almost disappear, and all nucleus are intensive
As can be seen from the above table: the rats in normal control group pancreas is normally, the rounded or oval-shaped cell mass of islets of langerhans, and sharpness of border, in islets of langerhans, the β cell is intensive, the rule that distributes, form is regular, and structure is normal, and endochylema is abundant.Model group animal insulin cell cytosol reduces and atrophy, and form is irregular, and number obviously reduces, the intensive imagination of nucleus, and the β cell size differs, and through Ridit check P<0.01, illustrates that the pancreas of modeling animal damages.With model group contrast, the metformin group can partly be resisted the beta Cell of islet atrophy, makes part endochylema be restored (P<0.01).Fructus Forsythiae lignanoid heavy dose of group cell number is more, and endochylema is abundant (P<0.01), in Fructus Forsythiae lignanoid, the small dose group phenomenon that can partly resist the atrophy of β cell cytosol has significant difference (P<0.01).
The impact of table 7 on rat diabetes model kidney due to streptozotocin
"-": glomerule, renal capsule, renal tubules are substantially all normal; "+": messangial cell is hypertrophy to some extent, and capsular space slightly has and narrows down, and renal cells is edema slightly, and tube chamber narrows down; " ++ ": the obvious hypertrophy of messangial cell, capsular space partly narrows down, the renal cells edema, tube chamber narrows down; " +++": the remarkable hypertrophy of messangial cell, capsular space narrows down or disappears, the obvious edema of renal cells, tube chamber narrows down or disappears
By upper table and pathology photo, can be found out: blank group basement membrane uniformity, without obviously thickening, the glomerular capillary chamber is without narrow, glomerule interstitial NIP cell invasion.Model group glomerular capillary chamber subsides, obturation, and glomerular volume increases, lymphocytic infiltration, basement membrane thickened and extracellular matrix increase; Renal cells swelling, in the kidney interstitial, large amount lymphocyte and mononuclear phagocyte infiltrate, and through Ridit check P<0.01, illustrate that the kidney of modeling animal damages.With model group contrast, metformin group mesonephric glomerulus, renal cells and interstitial normal (P<0.01).The heavy dose of group of Fructus Forsythiae lignanoid can alleviate extensive edema and the interstitial cell infiltration phenomenon (P<0.01) of renal cells; In Fructus Forsythiae lignanoid, small dose group can partly protect glomerule and renal tubules (P<0.01).
The impact of table 8 on rat diabetes model heart tissue due to streptozotocin
"-": myocardial cell is normal, without congested and inflammatory cell, invades profit; "+": the myocardial cell disorder, congested and invade profit without inflammatory cell; " ++ ": the disorderly thickness of myocardial cell differs, and nucleus increases, and congested and inflammatory cell is invaded profit; " +++": the disorderly obviously thickness of myocardial cell differs, the nucleus showed increased, obviously congested and inflammatory cell is invaded profit
By upper table and pathology photo, can be found out: blank treated animal myocardial cell marshalling, interstitial is without hyperemia and cell infiltration, all normal, compare model group myocardial cell arrangement disorder, cardiac muscle fiber atrophy with blank group, the obvious hyperemia of interstitial and cell infiltration, through the Ridit check, P<0.01, illustrate that pathological changes appears in the animal pattern heart.With model group, compare, metformin group myocardial cell marshalling, the myocardial cell atrophy is restored, and interstitial is without hyperemia and cell infiltration (P<0.01).Fructus Forsythiae lignanoid large, medium and small dosage group myocardial cell is arranged and is arranged, and part is resisted myocardial atrophy (P<0.01).
The impact of table 9 on rat diabetes model liver organization due to streptozotocin
"-": hepatocyte, sinus hepaticus are normal, without the congested inflammatory cell that squeezes, invade profit; "+": hepatocyte, sinus hepaticus are normal, have the congested inflammatory cell that squeezes to invade profit; " ++ "; The slight hepatic cell edema, sinus hepaticus narrows down, and has the congested inflammatory cell that squeezes to invade profit; " +++": hepatocyte edema and acidophile degeneration, sinus hepaticus narrows down or disappears, and has congested squeezing with inflammatory cell to invade profit
By upper table and pathology photo, can be found out: blank group hepatocyte is radial arrangement, and hepatic cords and sinus hepaticus are high-visible.With blank group, compare, around the central vein of model group liver of laboratory animal, hepatocyte is radial arrangement, the periphery arrangement disorder, part of hepatocytes is the acidophilia and changes, sinus hepaticus narrows down, and visible focal hepatic necrosis and cell infiltration, through Ridit, checks, P<0.01, illustrate that pathological changes appears in the animal pattern liver.With model group, compare, the hepatocyte of metformin group liver of laboratory animal is radial arrangement, hepatic cords and sinus hepaticus high-visible (P<0.01).The hepatocyte of Fructus Forsythiae lignanoid large, medium and small dosage group liver is radial arrangement, hepatic cords and sinus hepaticus high-visible (P<0.01).
The impact of table 10 on rat diabetes model lungs tissue due to streptozotocin
"-": around bronchioles, trachea, alveolar septum and alveolar all normal; "+": around bronchioles, trachea, alveolar septum mild hyperaemia and inflammatory cell invade profit and thicken, in alveolar without secretions; " ++ ": bronchioles epithelial cell Mild edema, around trachea, inflammatory cell is invaded profit, and the congested inflammatory cell of alveolar septum is invaded and is moistened and thicken, and alveolar is interior without secretions; " +++": the serious edema of bronchioles epithelial cell, around trachea, a large amount of inflammatory cells are invaded profit, and the congested inflammatory cell of alveolar septum is invaded and is moistened and thicken, and is full of a large amount of inflammatory cells in alveolar
By upper table and pathology photo, can be found out: bronchioles, respiratory bronchus, alveolar septum visible inflammatory cell in blank group lungs ooze out and congested phenomenon, and the broadening of lung interval is due to the long-time result that causes interstitial pneumonia of raising.With blank group, compare, extensive cell infiltration around bronchioles in the model group lungs, the visible a large amount of cell infiltration in alveolar and interval occur that alveolar disappears, the edema phenomenon appears in the bronchial mucosa epithelial cell, have no inflammatory exudation in tube chamber, without exudate, check through Ridit in alveolar, P<0.01, illustrate that pathological changes appears in the animal pattern lung.With model group, compare, the metformin group can partly be improved inflammatory cell in alveolar septum and ooze out and congested phenomenon (P<0.01).The large, medium and small dosage group of Fructus Forsythiae lignanoid all can be improved in various degree inflammatory cell in alveolar septum and oozes out and congested phenomenon, improves lung pathology situation (P<0.01).
By above-mentioned data, shown, Fructus Forsythiae lignanoid prepared by the present invention has the effect of blood sugar lowering, is effective to hypoglycemic medicine, has enlarged the medical value of Fructus Forsythiae, is the innovation in the medicine preparation, and good economic and social benefit is arranged.
Claims (2)
1. the preparation method of a Fructus Forsythiae extract lignanoid, it is characterized in that, Fructus Forsythiae powder was broken into to the medicated powder of 60-80 mesh sieve, the soak with ethanol 0.5h that for medicated powder, 8 times of weight, volumetric concentration are 80%, reflux, extract, 2 times, the 1st 1.5h, the 2nd 1h, filtering residue discards, and obtains extracting solution, and extracting solution is evaporated to without the alcohol flavor, adjust liquor strength and containing crude drug 0.5g/ml, obtain sample liquid for being equivalent to; AB-8 type macroporous adsorptive resins on sample liquid, the ratio of applied sample amount and resin volume is 1:12, the resin column blade diameter length ratio is 1:12, first uses the water elution of 8 times of column volumes, obtains the eluent I and discards; With 8 times of column volume, ethanol elutions that volumetric concentration is 10% of amount, obtain the eluent II and discard again; Finally, with 10 times of column volume, ethanol elutions that volumetric concentration is 75% of amount, obtain the eluent III, reclaim ethanol, the concentrating under reduced pressure drying, obtain Fructus Forsythiae lignanoid.
2. the application of Fructus Forsythiae lignanoid in preparing hypoglycemic medicine that the described method of claim 1 is extracted.
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