CN109045190A - A kind of Chinese medicine composition and its preparation method and application - Google Patents
A kind of Chinese medicine composition and its preparation method and application Download PDFInfo
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Abstract
The invention belongs to field of medicaments, in particular to a kind of Chinese medicine composition and its preparation method and application for treating diabetic keratopathy intracerebral minute lesion.Chinese medicine composition of the invention is made of using 9~15 parts by weight pheretimas, 9~15 parts by weight ramulus cinnamomi, 9~15 parts by weight Poria cocos, 9~15 parts by weight root barks of tree peony, 9~15 parts by weight radix paeoniae rubras, 9~15 parts by weight peach kernels, 9~15 parts by weight Radix Astragalis, 9~15 parts by weight Chinese yams, 9~15 parts by weight Radix Salviae Miltiorrhizaes, 9~15 parts by weight rhizoma anemarrhenaes, 9~15 parts by weight Cortex Phellodendris, 9~15 parts by weight Radix Paeoniae Albas as raw material.Preparation method includes: the Chinese medicine for weighing formula ratio, and water is added to decoct extraction together, merges and decocts extracting solution, is filtered, and is concentrated, and is centrifuged, dry, and dry extract is made.Chinese medicine composition of the invention can improve diabetic keratopathy intracerebral minute lesion patient's intracerebral minute lesion number, and have adjustment effect to its glycosylated hemoglobin and triglycerides, have prospect in medicine.
Description
Technical field
The invention belongs to field of medicaments, more particularly to a kind of Chinese medicine composition and its preparation method and application.
Background technique
The it is proposed of intracerebral minute lesion is derived from widened perivascular space (Virchow-Robin space, VRS), is
Finger is seen straight in the T2 weighting (T2WI) of head Magnetic resonance imaging (Magnetic Resonance Imaging, MRI)
The Radiologic imaging that diameter 3m m neat in edge below, boundary understand, change without ambient signals.Diabetes are kind to endanger pole
Big comprehensive disease, characterized by hyperglycemia caused by defect of insulin secretion and (or) insulin action obstacle.Continue height
Blood glucose and long-term metabolic disorder etc. can lead to body tissue's organ, the especially damage of heart and brain kidney and its dysfunction and failure,
It is the major reason for causing cerebrovascular events and occurring.In recent years, intracerebral minute lesion as cerebral apoplexy " precognition the factor " gradually by
To attention, intracerebral minute lesion is treated as " primary prevention " means of cerebral apoplexy and has become hot topic concerned by people, and
Diabetes are the common risk factors of cerebral apoplexy Yu intracerebral minute lesion, and diabetic cerebrovascular disease is also refractory difficult anti-disease
One of.For diabetic, by preventing diabetic keratopathy intracerebral minute lesion, further preventing brain stroke is very
It is necessary and feasible.
Therefore, make every effort to find always in clinical and experiment can be on the basis of glycemic control, further by the present inventor
It plays prevention or treats the drug of intracerebral minute lesion, and then the generation of preventing brain stroke.The present inventor consults domestic and foreign literature,
Definite effective drug is not found, the method that direct intervention means compare affirmative so far only has low dose of aspirin,
But for cerebrovascular disease caused by diabetes, function and effect are not satisfactory.
Summary of the invention
Present invention is mainly applied to combination of Chinese tradiational and Western medicine clinics --- and the people at highest risk of cerebrovascular disease (is especially associated with 2 types
The patient of diabetes), " primary prevention " medication as " cerebrovascular disease ".By taking the prevention of this medicine, " diabetic keratopathy intracerebral is micro-
The generation of small lesion ", and then the generation of prevention of cerebrovascular diseases.
Specifically, the present invention provides a kind of Chinese medicine composition for treating diabetic keratopathy intracerebral minute lesion, it is described in
Drug composition with 9~15 parts by weight pheretimas, 9~15 parts by weight ramulus cinnamomi, 9~15 parts by weight Poria cocos, 9~15 parts by weight root barks of tree peony, 9~
15 parts by weight radix paeoniae rubras, 9~15 parts by weight peach kernels, 9~15 parts by weight Radix Astragalis, 9~15 parts by weight Chinese yams, 9~15 parts by weight Radix Salviae Miltiorrhizaes,
9~15 parts by weight rhizoma anemarrhenaes, 9~15 parts by weight Cortex Phellodendris, 9~15 parts by weight Radix Paeoniae Albas are made of raw material.
In a preferred example, the Chinese medicine composition is prepared as a raw material with the Chinese medicine of following parts by weight: pheretima 10
Part, 10 parts of Chinese yam, 10 parts of Radix Salviae Miltiorrhizae, is known 10 parts of ramulus cinnamomi, 10 parts of Poria cocos, 10 parts of the root bark of tree peony, 10 parts of radix paeoniae rubra, 10 parts of peach kernel, 10 parts of Radix Astragali
Female 10 parts, 10 parts of Cortex Phellodendri, 10 parts of Radix Paeoniae Alba.
There is provided a kind of systems of Chinese medicine composition for treating diabetic keratopathy intracerebral minute lesion for the second aspect of the present invention
Preparation Method is made dry extract using water extraction using the Chinese medicine as raw material, includes the following steps:
A) pheretima, ramulus cinnamomi, Poria cocos, the root bark of tree peony, radix paeoniae rubra, peach kernel, Radix Astragali, Chinese yam, Radix Salviae Miltiorrhizae, rhizoma anemarrhenae, the Cortex Phellodendri of formula ratio are weighed
And Radix Paeoniae Alba；
B) it adds water to cook twice, decocts 1 hour every time, adding water body accumulated amount every time is 8 times of medicinal material total weight, is merged twice
Decocting liquid, filtering；
C) concentrating the filtrate to the relative density measured at room temperature is 1.20, is then 60%~95% with volume fraction
Ethanol water alcohol precipitation；
D) recycling ethyl alcohol and being concentrated into the relative density measured at room temperature is 1.36, obtains medicinal extract, is dried in vacuo, obtains dry leaching
Cream.
There is provided a kind of systems of Chinese medicine composition for treating diabetic keratopathy intracerebral minute lesion for the third aspect of the present invention
Preparation Method is made dry extract using alcohol extracting method using the Chinese medicine as raw material, includes the following steps:
A) pheretima, ramulus cinnamomi, Poria cocos, the root bark of tree peony, radix paeoniae rubra, peach kernel, Radix Astragali, Chinese yam, Radix Salviae Miltiorrhizae, rhizoma anemarrhenae, the Cortex Phellodendri of formula ratio are weighed
And Radix Paeoniae Alba；
B) plus volume fraction be 60%~95% ethanol water, refluxing extraction twice, 1.5 hours every time, each institute
With 8~10 times that the volume of ethanol water is medicinal material total weight；
C) merge alcohol extract, ethyl alcohol is recovered under reduced pressure, obtains medicinal extract, be dried in vacuo, obtain dry extract.
In a preferred example, the vacuum drying temperature of the medicinal extract is 60 DEG C~80 DEG C.
In another preferred example, the vacuum drying temperature of the medicinal extract is 70 DEG C.
The present invention also provides the Chinese medicine compositions in the drug of preparation treatment diabetic keratopathy intracerebral minute lesion
Using.
In a preferred example, the drug is granule, tablet, capsule, pill or pill.
The details of various aspects of the present invention will be able to detailed description in subsequent chapters.By hereafter and claim
Description, the features of the present invention, purpose and advantage will become apparent from.
Detailed description of the invention
The apoptosis rate of Fig. 1 each group nerve cell compares
*△Vs HGSCco P < 0.05, * *▲Vs NGSCco P < 0.05
NGSCco: normal to co-culture Sc group；HGSCco: high sugar co-cultures Sc group；HGCIco: high sugar compound group；HGSIco:
High sugar ramulus cinnamomi group；HGAIco: high sugar pheretima group；HGYIco: high sugar Radix Astragali group；
The expression of Fig. 2 each group schwann cell Bcl-2 albumen
△Vs HGSCco P < 0.05,▲Vs NGSCco P < 0.05,☆Vs HGCIco P < 0.05,※vs HGAIco P
< 0.05；
The expression of Fig. 3 each group schwann cell Casepase-3 albumen
△Vs HGSCco P < 0.05,▲Vs NGSCco P < 0.05,☆Vs HGCIco P < 0.05,※vs HGAIco P
< 0.05.
Specific embodiment
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.Reagent used in the following example and raw material can be bought by commercial sources to be obtained
?.In the following examples, the experimental methods for specific conditions are not specified, usually according to normal condition or according to proposed by manufacturer
Condition.Unless otherwise defined, it anticipates known to all professional and scientific terms as used herein and one skilled in the art
Justice is identical.In addition, any method similar to or equal to what is recorded and material can be applied to the method for the present invention.Wen Zhong
The preferred implement methods and materials are for illustrative purposes only.
The feature that the features described above or embodiment that the present invention mentions are mentioned can be in any combination.Patent specification is taken off
All features shown can be used in combination with any composition form, and each feature disclosed in specification any can provide phase
The alternative characteristics of same, impartial or similar purpose replace.Therefore except there is special instruction, revealed feature is only impartial or similar
The general example of feature.
Pheretima as used in the following examples, ramulus cinnamomi, Poria cocos, the root bark of tree peony, radix paeoniae rubra, peach kernel, Radix Astragali, Chinese yam, Radix Salviae Miltiorrhizae, rhizoma anemarrhenae,
Cortex Phellodendri, Radix Paeoniae Alba meet related every regulation under 2005 editions each medicinal material items of a text of Chinese Pharmacopoeia.Each medicinal material has been subjected to only
The working processes such as system, cutting, processing, crushing, before feeding intake, by identification, ingredients material object is consistent with title, and quality meets state
Family's standards of pharmacopoeia (specific Med Mat Appreciation method is executed referring to standards of pharmacopoeia).
Embodiment 1
Weigh pheretima 10g, ramulus cinnamomi 10g, Poria cocos 10g, root bark of tree peony 10g, radix paeoniae rubra 10g, peach kernel 10g, Radix Astragali 10g, Chinese yam 10g,
Radix Salviae Miltiorrhizae 10g, rhizoma anemarrhenae 10g, Cortex Phellodendri 10g, Radix Paeoniae Alba 10g；
It adds water to cook twice, decocts 1 hour every time, add water 216ml every time；Merge decocting liquid twice, filtering；
Concentrating the filtrate to the relative density measured at 25 DEG C is 1.20, and the ethyl alcohol for being then 70% with volume fraction is water-soluble
Liquid carries out alcohol precipitation；
Recycling ethyl alcohol and being concentrated into the relative density measured at 25 DEG C is 1.36, and finally vacuum is dry under conditions of 70 DEG C
It is dry, obtain dry extract 4g.
Embodiment 2
Weigh pheretima 15g, ramulus cinnamomi 15g, Poria cocos 15g, root bark of tree peony 15g, radix paeoniae rubra 15g, peach kernel 15g, Radix Astragali 15g, Chinese yam 15g,
Radix Salviae Miltiorrhizae 15g, rhizoma anemarrhenae 15g, Cortex Phellodendri 15g, Radix Paeoniae Alba 15g；
It adds water to cook twice, decocts 1 hour every time, add water 216ml every time；Merge decocting liquid twice, filtering；
Concentrating the filtrate to the relative density measured at 25 DEG C is 1.20, and the ethyl alcohol for being then 70% with volume fraction is water-soluble
Liquid carries out alcohol precipitation；
Recycling ethyl alcohol and being concentrated into the relative density measured at 25 DEG C is 1.36, and finally vacuum is dry under conditions of 70 DEG C
It is dry, obtain dry extract 4g.
Embodiment 3
It weighs pheretima 9g, ramulus cinnamomi 9g, Poria cocos 9g, root bark of tree peony 9g, radix paeoniae rubra 9g, peach kernel 9g, Radix Astragali 9g, Chinese yam 9g, Radix Salviae Miltiorrhizae 9g, know
Female 9g, Cortex Phellodendri 9g, Radix Paeoniae Alba 9g；
It adds water to cook twice, decocts 1 hour every time, add water 216ml every time；Merge decocting liquid twice, filtering；
Concentrating the filtrate to the relative density measured at 25 DEG C is 1.20, and the ethyl alcohol for being then 70% with volume fraction is water-soluble
Liquid carries out alcohol precipitation；
Recycling ethyl alcohol and being concentrated into the relative density measured at 25 DEG C is 1.36, and finally vacuum is dry under conditions of 70 DEG C
It is dry, obtain dry extract 4g.
Embodiment 4
Weigh pheretima 15g, ramulus cinnamomi 10g, Poria cocos 10g, root bark of tree peony 10g, radix paeoniae rubra 10g, peach kernel 10g, Radix Astragali 30g, Chinese yam 15g,
Radix Salviae Miltiorrhizae 10g, rhizoma anemarrhenae 10g, Cortex Phellodendri 10g, Radix Paeoniae Alba 10g；
It adds water to cook twice, decocts 1 hour every time, add water 216ml every time；Merge decocting liquid twice, filtering；
Concentrating the filtrate to the relative density measured at 25 DEG C is 1.20, and the ethyl alcohol for being then 70% with volume fraction is water-soluble
Liquid carries out alcohol precipitation；
Recycling ethyl alcohol and being concentrated into the relative density measured at 25 DEG C is 1.36, and finally vacuum is dry under conditions of 70 DEG C
It is dry, obtain dry extract 4g.
Embodiment 5
Weigh pheretima 15g, ramulus cinnamomi 10g, Poria cocos 10g, root bark of tree peony 10g, radix paeoniae rubra 10g, peach kernel 10g, Radix Astragali 30g, Chinese yam 15g,
Radix Salviae Miltiorrhizae 15g, rhizoma anemarrhenae 10g, Cortex Phellodendri 10g, Radix Paeoniae Alba 10g；
It adds water to cook twice, decocts 1 hour every time, add water 216ml every time；Merge decocting liquid twice, filtering；
Concentrating the filtrate to the relative density measured at 25 DEG C is 1.20, and the ethyl alcohol for being then 70% with volume fraction is water-soluble
Liquid carries out alcohol precipitation；
Recycling ethyl alcohol and being concentrated into the relative density measured at 25 DEG C is 1.36, and finally vacuum is dry under conditions of 70 DEG C
It is dry, obtain dry extract 4g.
Relative density is 1.4；It is centrifuged 1 minute under the revolving speed of 4000r/min；It is dried in vacuo at 70 DEG C, obtains dry extract.
Embodiment 6
Weigh pheretima 150g, ramulus cinnamomi 100g, Poria cocos 100g, root bark of tree peony 100g, radix paeoniae rubra 100g, peach kernel 100g, Radix Astragali 300g, mountain
Medicine 150g, Radix Salviae Miltiorrhizae 150g, rhizoma anemarrhenae 100g, Cortex Phellodendri 100g, Radix Paeoniae Alba 100g；
It adds water to cook twice, decocts 1 hour every time, add water 28.8L every time；Merge decocting liquid twice, filtering；
Concentrating the filtrate to the relative density measured at 25 DEG C is 1.20, and the ethyl alcohol for being then 70% with volume fraction is water-soluble
Liquid carries out alcohol precipitation；
Recycling ethyl alcohol and being concentrated into the relative density measured at 25 DEG C is 1.36, and finally vacuum is dry under conditions of 70 DEG C
It is dry, obtain dry extract 480g.
Embodiment 7
Weigh pheretima 300g, ramulus cinnamomi 100g, Poria cocos 100g, root bark of tree peony 100g, radix paeoniae rubra 100g, peach kernel 100g, Radix Astragali 300g, mountain
Medicine 150g, Radix Salviae Miltiorrhizae 150g, rhizoma anemarrhenae 100g, Cortex Phellodendri 100g, Radix Paeoniae Alba 150g；
It adds water to cook twice, decocts 1 hour every time, add water 28.8L every time；Merge decocting liquid twice, filtering；
Concentrating the filtrate to the relative density measured at 25 DEG C is 1.20, and the ethyl alcohol for being then 90% with volume fraction is water-soluble
Liquid carries out alcohol precipitation；
Recycling ethyl alcohol and being concentrated into the relative density measured at 25 DEG C is 1.36, and finally vacuum is dry under conditions of 80 DEG C
It is dry, obtain dry extract 480g.
Embodiment 8
Weigh pheretima 200g, ramulus cinnamomi 100g, Poria cocos 100g, root bark of tree peony 150g, radix paeoniae rubra 150g, peach kernel 100g, Radix Astragali 300g, mountain
Medicine 150g, Radix Salviae Miltiorrhizae 150g, rhizoma anemarrhenae 100g, Cortex Phellodendri 100g, Radix Paeoniae Alba 150g；
It adds water to cook twice, decocts 1 hour every time, add water 28.8L every time；Merge decocting liquid twice, filtering；
Concentrating the filtrate to the relative density measured at 25 DEG C is 1.20, and the ethyl alcohol for being then 70% with volume fraction is water-soluble
Liquid carries out alcohol precipitation；
Recycling ethyl alcohol and being concentrated into the relative density measured at 25 DEG C is 1.36, and finally vacuum is dry under conditions of 60 DEG C
It is dry, obtain dry extract 460g.
Embodiment 9
It takes dry extract 150g (being equivalent to crude drug 1000g), starch 80g, dextrin 120g, sodium carboxymethyl starch 20g is added, with
50% ethyl alcohol crosses the pelleting of 14 meshes as adhesive, softwood processed, dry, and appropriate talcum powder tabletting, plain piece packet film is added
Clothing is to get using the tablet of Chinese medicine composition preparation.
Embodiment 10
Take the total 230g of dry extract, lactose 480g, mannitol 100g, cross-linked pvp 100g, 95% appropriate amount of ethanol, magnesium stearate
15g.The tabletted Solide dispersion tablets to get use Chinese medicine composition preparation.
Embodiment 11
Dry extract 200g is taken, Macrogol 4000 is added in 1:2 ratio, mixes, heating melting, closed and heat preservation is 80
DEG C, it instills in 0~10 DEG C of atoleine, collects dripping pill, drip is most and wipes coolant liquid to get using the Chinese medicine composition to prepare
Dripping pill.
Embodiment 12
Dry extract 200g, lactose 90g, magnesium stearate 1.5g are taken, is pressed after mixing large stretch of (slice weight 2g), this pre- tabletting is being shaken
It is crushed in pendulum-type granule, whole grain, is dispensed with aluminum-plastic composite membrane to get using the granule of Chinese medicine composition preparation.
Embodiment 13
Dry extract 200g is taken, starch 150g is added, is mixed, No. seven sieves are crossed, it is encapsulated to get using the Chinese medicine composition
The capsule of preparation.
Embodiment 14
20g beeswax, 500g soybean oil are heated to about 50 DEG C, stirring melts completely to it, that is, beeswax oil is made；Take dry leaching
Cream 500g is added in beeswax oil, stirs evenly；It is sieved with colloid mill wet pulverizing, oil suspension is made, it is spare；300g gelatin is taken to add
Enter suitable quantity of water, be allowed to water swelling, will be added in the gelatin expanded after the heating of suitable 150g glycerol, be heated with stirring to melting；It will
Shell glue filtering suction removes part moisture content and foam, using rotary-die type soft capsule manufacturing machine, by deployed content pressure
Enter shell film, is wrapped in and wherein forms soft capsule.The greasy dirt for washing away capsule surface, being placed in temperature is 25-30 DEG C, and relative humidity is
It dries in the baking oven of 20-25% to get using the soft capsule of Chinese medicine composition preparation.
Embodiment 15
Weigh pheretima 15g, ramulus cinnamomi 10g, Poria cocos 10g, root bark of tree peony 10g, radix paeoniae rubra 10g, peach kernel 10g, Radix Astragali 30g, Chinese yam 15g,
Radix Salviae Miltiorrhizae 10g, rhizoma anemarrhenae 10g, Cortex Phellodendri 10g, Radix Paeoniae Alba 10g；
Original prescription granule mixture 5.0%, maltodextrin 93.9%, mixed colorant (chocolate palm fibre: orange: it is white:
Brilliant blue=4.4:3.2:2.5:0.1) 1.0%, bitter principle 0.1%.It is total: 100%.First will by sieve powder process according to above-mentioned prescription
Mixed colorant adds water to be mixed evenly, and adds soluble starch, is mixed evenly.By mixture hand over dry medicine process into
Row drying.Mixture after drying is crushed, is uniformly mixed.Said mixture is pelletized, the coloring agent particle moisture requirement
Lower than 6.5%.It sieves powder process and gets granule according to above-mentioned granulate mixture prescription, weigh the corresponding prescription of each kind and use
Amount, gets the colored particles of moisture qualification, is uniformly mixed with granulate mixture, is packed into polyethylene plastic bag.
Carry out the beneficial effect of the present invention is further explained the drug below by clinical test example, the side of tearing open test illustrates
The beneficial effect of compound formulation.
Embodiment 16
Clinical trial data
1 clinical data and method
1.1 clinical data
Selection our hospital's Diabetic Inpatients 60, checking confirmation through head magnetic resonance (MRI), there are the small diseases of intracerebral
Become, knows, voluntarily participates in this research and sign informed consent form.According to entering, group time is randomly divided into observation group and control group is each
30.Observation group male 17, female 13；Year at age (70.0 ± 5.2).Control group male 16, female 14；Age (69.63 ±
5.1) year.Two groups of ages gender, smoke, minute lesion quantity, glycosylated hemoglobin, three in medical history of drinking and treatment treatment forebrain
Acyl glycerol liquor mean difference is not significant (P > 0.05), is comparable.
1.2 treatment method
Two groups are given identical Primary Care, including control blood pressure, control blood glucose etc..Medicine of the invention is taken by treatment group
Compositions (oral liquid formulations prepared by embodiment 1), 2 times/day.Control group gives aspirin on the basis of Primary Care
Enteric coatel tablets 0.1g 1 time every night, takes orally.Two groups equal 3 months be 1 course for the treatment of, two groups observe 2 courses for the treatment of.
1.3 are included in and exclusion criteria
1.3.1 be included in standard (1) clinical criteria: the age 18~80 years old, clinic was clarified a diagnosis as diabetes, head B-sonography inspection
It looks into there are intracerebral minute lesion and without corresponding symptom and positioning sign.2. iconography standard: while the T1 for shooting head B-sonography adds
Weigh (T1WI), T2 weights (T2WI), fluid attented inversion recovery picture (FLAIR).Intracerebral minute lesion is defined as on T2WI directly
Diameter < 3mm, neat in edge, boundary is clear, changes without surrounding, the uniform high RST domain of signal.The trouble of minute lesion >=6
Person significantly increases compared with≤5 person's Cerebral Infarction rates, so this research, which is mainly chosen in Basal ganglia 2/3, shows as the low letter of T1WI
Number, the signals (II type) such as T2WI high RST, FLAIR and T1WI low signal, T2WI high RST, FLAIR low signal (III type) it is micro-
Small lesion, and the multiple minute lesion case of two sides Basal ganglia >=6 total (auspicious safe 2 grades of classification of holt or more).
1.3.2 there are lacunar infarction or other infarcts persons for exclusion criteria (1) head B-sonography inspection；(2) exist when going to a doctor
(i.e. the lesion meets minute lesion in iconography, but has corresponding disease by the corresponding symptom of responsibility lesions and positioning sign person
Shape and positioning sign)；(3) to drug ingedient (aspirin or pheretima) allergy sufferers in this research；(4) tumor patient or with
Serious conscience kidney diaseases, digestive tract ulcer, gout have hemorrhagic tendency person；(5) gestation or nursing women；(6) it participates in the recent period
Cross other clinical test persons；(7) used aspirin or other antiplatelets, anticoagulant person are taken within nearly 3 months orally.Experimental period
Between, patient in group such as occurs serious adverse events or unexpected pregnancy, suffers from malignant tumour or other malignant diseases and other occur
Test is exited when may influence the disease of observation result.
1.4 observation index (1) overview project: respectively when entering group and after treatment 3 months, 6 months, follow-up observation brain
The relevant risk factor situation of interior minute lesion mainly includes age, gender, Yin Jiu ﹑ smoking, hypertension (systolic pressure, diastole
Pressure), hyperlipidemia (serum total cholesterol, triacylglycerol, high-density lipoprotein cholesterol, low density lipoprotein cholesterol), and
Record patient blood glucose's situation (glycosylated hemoglobin including empty stomach and 2h-plasma glucose) in detail.(2) curative effect index: enter group 3
Month, after 6 months, check the head B-sonography that shoots under identical conditions, observing the situation of change of intracerebral minute lesion, (including intracerebral is micro-
Increase and decrease, the change of form and the variation of phenotype of minor illness parameter amount).Intracerebral minute lesion treatment effectiveness evaluation: minute lesion
It reduces to 5 the following are effective, quantity, form do not become effectively, and it is invalid that quantity, form, which increase,.
1.5 statistical procedures carry out statistical procedures using SPSS18.0 software, and measurement data is using single factor test variance point
Analysis or t are examined, and ranked data use rank sum test, and it is significant difference with P < 0.05 that enumeration data, which uses chi-square criterion,.
2 results
Follow-up 6 months, two groups of nothings exited case；Do not occur adverse reaction relevant to drug during treatment, enters a group disease
Example pretherapy and post-treatment blood, urine, feces routine and hepatic and renal function are without significant changes.
2.1 two groups of pretherapy and post-treatment intracerebral minute lesion quantity compare the preceding two groups of intracerebrals minute lesion quantity variance for the treatment of and do not show
It writes (P > 0.05).3 months, 6 months are treated, before two groups of intracerebral minute lesion quantity are substantially less than the treatment of this group；Treatment 6
Month, observation group's intracerebral minute lesion quantity is substantially less than control group (P < 0.05).It is shown in Table 1.
Minute lesion quantity compares before 1 two groups of treatments of table and in treatment March, June hindbrain
Note: with this group before treatment, * P < 0.05；Compared with control group is with timing node, △ P < 0.05
Before 2.2 two groups of pretherapy and post-treatment glycosylated hemoglobins and serum triglyceride level compare treatment, two groups of glycosylated hemoglobins,
The equal difference of serum triglyceride level is not significant (P > 0.05).Treat 3 months, 6 months, two groups of glycosylated hemoglobins, triacylglycerol water
Before being averagely substantially less than the treatment of this group；Treatment 6 months, observation group's glycosylated hemoglobin, serum triglyceride level, which are substantially less than, to be compareed
Group (P < 0.05).It is shown in Table 2.
2 two groups of pretherapy and post-treatment glycosylated hemoglobins of table and serum triglyceride level compare
Note: with this group before treatment, * P < 0.05；Compared with control group is with timing node, △ P < 0.05
3 conclusions: pharmaceutical composition of the invention can improve diabetic keratopathy intracerebral minute lesion patient's intracerebral minute lesion number
Mesh, and have adjustment effect to its glycosylated hemoglobin and triglycerides.
The pharmaceutical composition side of tearing open of the invention of embodiment 17 is to neuroprotective function experimental study
1 material and method
1.1 trial drug pheretima water decoctions；Ramulus cinnamomi, Poria cocos, the root bark of tree peony, radix paeoniae rubra, peach kernel water decoction；Radix Astragali, Radix Salviae Miltiorrhizae, Chinese yam,
Rhizoma anemarrhenae, Cortex Phellodendri, Radix Paeoniae Alba water decoction, by the offer of Pharmacy of Shanghai City Hospital of Combination of Chinese Traditional and Western Medicin, (every 1ml extracting solution is equivalent to 2g original
Medicine).
The SD rat that 1.2 animals are born 1-3 days, adult SD rats weight 150-200g, male and female are unlimited, by Fudan University
Extra large laboratory animal center of medical college provides.
1.3 reagent Primary antibodies: rabbit-anti rat Bcl-2Antibody, rabbit-anti rat Casepase-3Antibody are purchased from
CellSignaling company；The anti-GAPDH polyclonal antibody of mouse is provided by upper Haikang at Reagent Company；Secondary antibody: HRP mark
Remember goat anti-rabbit igg, HRP label goat anti-mouse IgG has upper Haikang to provide at Reagent Company.
2 statistical dispositions
The data obtained carries out statistical disposition with SPSS20.0 software.Measurement data uses " mean ± standard deviation "It indicates, meets normal distribution person, mean difference t inspection or variance analysis between each group；Non-Gaussian Distribution person uses order
And inspection, P < 0.05 think there is statistical difference.
3 experimental methods carry out the culture, identification and passage of rat nerve cell with suckling mouse nerve, by newborn 3dSD rat 8-
10 sacrificed by decapitation, sterile lower separation bilateral sciatic nerve and brachial plexus nerve, remove its perilemma epineurium, and shred nerve, use glue
Protoenzyme digestion after be centrifuged, add 10%FBS-DMEM culture solution (5.5mmol/L glucose), 37 DEG C, the culture of 5%CO2 incubator for 24 hours
After change Ara-c cell culture fluid, be further cultured for replacing cell culture fluid afterwards for 24 hours, change weekly later liquid 2~3 times.Exempted from S100 antibody
Epidemic disease groupization detects, and occurs sepia positive particle person in endochylema and is accredited as schwann cell, the schwann cell that this method obtains is pure
Degree up to 95%.It is passed on trypsin digestion, tests the 2nd, 3 generation cells.
Heparin sodium is injected intraperitoneally by adult SD rats 1 in the culture for carrying out rat endothelial cells with digestion method simultaneously
6250U, sterile lower separation and clip aorta pectoralis, the outer connective tissue of removal blood vessel and outer membrane, draw 4 DEG C of PBS with 1ml syringe
Lumen of vessels is rinsed, remaining haemocyte is removed.The tiny branch that aorta issues is removed with eye scissors, blood vessel is cut into thick about
1.5mm vascular circle.PBS washes out vascular circle after incubator stands 60 hours, adds the 10%FBS-DMEM culture solution (Portugal 5.5mmol/L
Grape sugar), 37 DEG C, 5%CO2 incubator culture change liquid in every 2~3 days later.With VIII factor immunofluorescence assay, fluorescence in endochylema
Colour developing person is accredited as endothelial cell, and positive rate is greater than 90%.It is passed on trypsin digestion, tests the 2nd, 3 generation cells.
After cultivating successfully, the co-cultivation of schwann cell and endothelial cell is carried out.It co-cultures successfully, by grouping experiment: high Portugal
Grape sugar adds pheretima intervention group (referred to as, high sugar pheretima group, HGAIco): liquid is changed after ibid co-culturing, be added 25mM containing glucose,
The DMEM culture solution of imperial 8mg/ml.Referred to as, high glucose adds guizhi fuling pill intervention group (high sugar ramulus cinnamomi group, HGSIco): ibid
Liquid is changed after co-cultivation, and the DMEM culture solution of 25mM containing glucose, guizhi fuling pill 4mg/ml is added.High glucose adds the medicines such as Radix Astragali
Object group intervention group (referred to as, high sugar Radix Astragali group, HGYIco): liquid is changed after ibid co-culturing, 25mM containing glucose, Radix Astragali etc. is added
The DMEM culture solution of 8mg/ml.High glucose adds compound intervention group (referred to as, high sugar compound group, HGCIco): changing after same co-cultivation
The DMEM culture solution of 25mM containing glucose, compound (above three groups of drug mixture 8mg/ml) is added in liquid.
Observe nerve cell apoptosis rate, Bcl-2mRNA and protein expression, Casepase 3mRNA and protein expression.
4. intervention effect of the different Chinese medicines to the nerve cell apoptosis co-cultured caused by high sugar
The measurement of 4.1 nerve cell apoptosis rates:
Propidium iodide (PI) flow cytomery apoptosis rate, normal apoptosis rate is almost nil, compared with normal group
High sugar co-cultures Sc group apoptosis rate and obviously increases 2.11 (P < 0.05)；Compared with high sugar co-cultures Sc group, group of cells apoptosis rate
It is all substantially reduced (P < 0.05), wherein with high sugared compound group apoptosis rate minimum 0.32, (P > 0.05).(see Fig. 1, table 3)
Table 3
4.2 cloning Bcl-2 protein expressions
High sugar co-cultivation Sc group bcl-2 expression quantity is obvious compared with normally organizing it can be seen from Western blot result
It reduces (P < 0.05), normal group is 1.69, and it is 0.30 that high sugar, which co-cultures Sc group,；Compared with high sugar co-cultures Sc group, each group bcl-
2 expression quantity all increased (P < 0.05), wherein remaining is each with high sugared compound group bcl-2 incrementss most 0.76 (P > 0.05)
Significant difference between group.(such as Fig. 2, table 4)
Table 4
4.3 Apoptosis silver Casepase-3 protein expressions
High sugar co-cultures Sc group, casepase-3 expression compared with normally organizing it can be seen from Western blot result
Amount increases obvious (P < 0.05), and normal group is 0.075, and it is 0.53 that high sugar, which co-cultures Sc group,；Compared with high sugar co-cultures Sc group,
Each group Casepase-3 expressing quantity gene expression amount all significantly reduces (P < 0.05), height sugar compound group 0.095 (P >
0.05), significant difference between remaining each group.(such as Fig. 3, table 5)
Table 5
5 the results shows: our medicament composing prescription has protective effect to the damage of nerve cell, wherein with compound
Protective effect is better than the respectively side's of tearing open group, has synergistic function between each drug in yin-nourishing eliminating mass side.
Claims (8)
1. a kind of Chinese medicine composition for treating diabetic keratopathy intracerebral minute lesion, which is characterized in that the Chinese medicine composition with 9~
15 parts by weight pheretimas, 9~15 parts by weight ramulus cinnamomi, 9~15 parts by weight Poria cocos, 9~15 parts by weight root barks of tree peony, 9~15 parts by weight radix paeoniae rubras,
9~15 parts by weight peach kernels, 9~15 parts by weight Radix Astragalis, 9~15 parts by weight Chinese yams, 9~15 parts by weight Radix Salviae Miltiorrhizaes, 9~15 parts by weight are known
Mother, 9~15 parts by weight Cortex Phellodendris, 9~15 parts by weight Radix Paeoniae Albas are made of raw material.
2. Chinese medicine composition as described in claim 1, which is characterized in that the Chinese medicine composition is with the Chinese medicine of following parts by weight
Material is made of raw material: 10 parts of pheretima, 10 parts of ramulus cinnamomi, 10 parts of Poria cocos, 10 parts of the root bark of tree peony, 10 parts of radix paeoniae rubra, 10 parts of peach kernel, 10 parts of Radix Astragali,
10 parts of Chinese yam, 10 parts of Radix Salviae Miltiorrhizae, 10 parts of rhizoma anemarrhenae, 10 parts of Cortex Phellodendri, 10 parts of Radix Paeoniae Alba.
3. a kind of preparation method of Chinese medicine composition as described in claim 1, which is characterized in that using the Chinese medicine as raw material
Dry extract is made using water extraction, is included the following steps:
A) pheretima of formula ratio, ramulus cinnamomi, Poria cocos, the root bark of tree peony, radix paeoniae rubra, peach kernel, Radix Astragali, Chinese yam, Radix Salviae Miltiorrhizae, rhizoma anemarrhenae, Cortex Phellodendri and white are weighed
Chinese herbaceous peony；
B) it adds water to cook twice, decocts 1 hour every time, adding water body accumulated amount every time is 8 times of medicinal material total weight, merges decocting twice
Liquid, filtering；
C) concentrating the filtrate to the relative density measured at room temperature is 1.20, the ethyl alcohol for being then 60%~95% with volume fraction
Aqueous solution alcohol precipitation；
D) recycling ethyl alcohol and being concentrated into the relative density measured at room temperature is 1.36, obtains medicinal extract, is dried in vacuo, obtains dry extract.
4. a kind of preparation method of Chinese medicine composition as described in claim 1, which is characterized in that using the Chinese medicine as raw material
Dry extract is made using alcohol extracting method, is included the following steps:
A) pheretima of formula ratio, ramulus cinnamomi, Poria cocos, the root bark of tree peony, radix paeoniae rubra, peach kernel, Radix Astragali, Chinese yam, Radix Salviae Miltiorrhizae, rhizoma anemarrhenae, Cortex Phellodendri and white are weighed
Chinese herbaceous peony；
B) plus volume fraction be 60%~95% ethanol water, refluxing extraction twice, 1.5 hours every time, second used every time
The volume of alcohol solution is 8~10 times of medicinal material total weight；
C) merge alcohol extract, ethyl alcohol is recovered under reduced pressure, obtains medicinal extract, be dried in vacuo, obtain dry extract.
5. the preparation method of Chinese medicine composition as described in claim 3 or 4, which is characterized in that the medicinal extract is vacuum drying
Temperature is 60 DEG C~80 DEG C.
6. the preparation method of Chinese medicine composition as claimed in claim 5, which is characterized in that the vacuum drying temperature of medicinal extract
It is 70 DEG C.
7. Chinese medicine composition as described in claim 1 answering in the drug of preparation treatment diabetic keratopathy intracerebral minute lesion
With.
8. the use as claimed in claim 7, which is characterized in that the drug be granule, tablet, capsule, pill or
Pill.
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| CN109224067A (en) * | 2018-10-24 | 2019-01-18 | 上海市中西医结合医院 | A kind of production method of diabetic keratopathy intracerebral minute lesion model rodent |
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| CN104337884A (en) * | 2013-08-06 | 2015-02-11 | 颜明 | Medicine composition for preventing and/or treating diabetes and complication of diabetes |
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| CN109224067A (en) * | 2018-10-24 | 2019-01-18 | 上海市中西医结合医院 | A kind of production method of diabetic keratopathy intracerebral minute lesion model rodent |
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