CN101912565B - Turmeric and astragalus capsules for treating diabetes and incipient diabetic nephropathy - Google Patents
Turmeric and astragalus capsules for treating diabetes and incipient diabetic nephropathy Download PDFInfo
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Abstract
The invention relates to turmeric and astragalus capsules for treating diabetes and incipient diabetic nephropathy and can effectively solve the problem of treating the diabetes and the incipient diabetic nephropathy. The turmeric and astragalus capsules are prepared from 105 to 130 grams of turmeric extract, 65 to 85 grams of astragalus extract (mainly total saponins of astragalus (ASTs)), starch and talcpowder by the steps of grinding the turmeric extract and the astragalus extract, sieving powder through a screen of 100 meshes, uniformly mixing the powder, adding the starch and the talcpowder into the mixture to uniformly mix, carrying out granulation on the mixture and encapsulating, wherein the adding amount of the starch and the talcpowder is less than or equal to 10 percent of the total weight of the turmeric extract, the astragalus extract, the starch and the talcpowder; and the weight ratio of the starch to the talcpowder is approximately 1:1. The turmeric and astragalus capsules have the advantages of simple preparation method, suitable compatibility, good curative effect, no toxic or side effects, obvious effect of treating the diabetes and the incipient diabetic nephropathy, innovation of treating the diabetes and the incipient diabetic nephropathy and enormous economic and social benefits.
Description
One, technical field
The present invention relates to the field of Chinese medicines, particularly a kind of Rhizoma Zingiberis Recens astragalus capsules of treating diabetes and early diabetes nephropathy.
Two, background technology
Diabetes are the quick diseases that increase of sickness rate in the world, and according to the statistics of World Health Organization (WHO), whole world diabetes number was 1.35 hundred million in 1998, reached 1.6 hundred million, 2004 and reached 2.4 hundred million in 2000.Expecting the year two thousand thirty will reach 3.7 hundred million.First three country that whole world diabetes number of patients is maximum is India, China and the U.S..The prevalence of China's diabetes is more than 3% at present, and the number of patients of diabetes is estimated to have reached about 5,000 ten thousand.The diabetes sickness rate of each stratum throughout the country are all very high, become the fourth-largest killer who threatens human health.
Diabetes fearful also do not lie in diabetes itself, and is its complication.The complication of diabetes is harmful to patient's long-term health, influences quality of life and life expectancy, is that the diabetes patient is disabled and main causes of death.(DiabeticNephropathy DN) is meant that (Diabetes mellitus, the DM) change of the caused kidney 26S Proteasome Structure and Function of microangiopathies are one of common and serious long term complications of DM to diabetes to diabetic nephropathy.Along with the change of raising, aged tendency of population and the life style of living standards of the people, in the world wide, the DM sickness rate is increasing, and DN also sharply increases thereupon.(about 40% ESRD patient is by due to the DN for End stage renal disease, main cause ESRD), is to be caused by DN more than 50% among the annual newly-increased ESRD patient to have become nephropathy in latter stage at end at western countries DN.The DM prevalence also constantly increases in China, and China DM patient surpasses 40,000,000 according to estimates.China also rises because of the number that chronic renal failure takes place DN year by year.In a single day the renal damage performance takes place in patient DM, and the persistence urine protein is difficult to reverse, and finally proceeds to renal failure in latter stage at end.Therefore how preventing in early days that DM from developing to DN, or adopt effective measures to stop DN to make progress to whole latter stage, is one of focus of Chinese scholars research.
At present modern medicine for diabetic complication particularly diabetic nephropathy still lack effective medicine; Modern medicine will be later than the traditional Chinese medical science to the understanding of diabetes; By anatomy and biochemistry, pathological research method; Recognize the inherent pathological change and the Clinical types of diabetes, along with the synthetic of insulin and chemosynthesis antidiabetic drug being widely used clinically, the glycemic control of diabetes in most of the cases can realize more clearlyly.But simple blood sugar reducing function is to the many no clear and definite curative effects of complication, and the easy drug resistance that produces.The patient who much takes the treatment of antidiabetic drug and insulin injection is along with the prolongation of time, and dose can increase gradually, and the drug kinds of input also often gradually becomes associating by single; Prolonged application irritates islets of langerhans, makes pancreas be in nervous duty for a long time, finally causes the islet function obstacle; Occur blood glucose simultaneously and rebound repeatedly, hemorheology changes, and forming the blood stickiness increases; Blood microcirculation generation obstacle; Cause the heart, brain, kidney complication and weight increase at last, edema, untoward reaction such as liver drug enzyme rising.
Three, summary of the invention
To above-mentioned situation, for overcoming the defective of prior art, the object of the invention just is to provide a kind of Rhizoma Zingiberis Recens astragalus capsules of treating diabetes and early diabetes nephropathy; Can effectively solve the problem of diabetes and early diabetes treatment of kidney disease; The present invention is processed by Rhizoma Curcumae Longae extract 105～130g, Radix Astragali extract (mainly being Radix Astragali total saponins) 65～85g, starch, Pulvis Talci, and Rhizoma Curcumae Longae extract and Radix Astragali extract were pulverized 100 mesh sieves, mix homogeneously; Add starch again, Pulvis Talci is mixed; Granulate, incapsulate, promptly get;
Starch, Pulvis Talci addition≤Rhizoma Curcumae Longae extract, Radix Astragali extract, starch, Pulvis Talci gross weight and 10%, starch, Pulvis Talci weight ratio are about 1: 1.
Said Rhizoma Curcumae Longae extract is, takes by weighing the Rhizoma Curcumae Longae medical material after the pulverizing, puts into microwave extraction tank; Amount of ethanol is 20 times of Rhizoma Curcumae Longae medical material weight, and promptly Rhizoma Curcumae Longae and alcoholic acid weight ratio are 1: 20, extracts 50 ℃ of temperature; Extraction time 30min extracts power 200W, and it is subsequent use to get the Rhizoma Curcumae Longae crude extract;
The S-8 purification with macroreticular resin: the S-8 macroporous adsorbent resin is earlier with soak with ethanol 24 hours, is washed with distilled water to that not have alcohol afterwards subsequent use; Macroporous adsorbent resin wet method dress post with handling well adsorbs with the 1mL/min flow velocity Rhizoma Curcumae Longae crude extract through chromatographic column, elder generation is with the water elution impurity of 4 times of resin volumes, the dehydrated alcohol eluting of 8 times of resin volumes of reuse; Collect the dehydrated alcohol eluent, concentrating under reduced pressure ethanol, drying and crushing; Rhizoma Curcumae Longae extract, yield is about 3.5%, these article are xanchromatic powder; Bitter in the mouth, spectrophotometry, these article contain total curcumin and are no less than 58.0% in curcumin; The HPLC method is measured, and the every 1g of these article contains Rhizoma Curcumae Longae in curcumin (C21H20O 6), must not be less than 15.0%.
Total curcumin: measure according to spectrophotography (appendix V of Chinese Pharmacopoeia version in 2005), these article contain total curcumin and are no less than 58.0% in curcumin.
Curcumin: measure according to HPLC (an appendix VI of Chinese Pharmacopoeia version in 2005 D).Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filler; Mobile phase is acetonitrile-0.5% aqueous acetic acid (45: 55), flow velocity 1.0mLmin
-1, detect wavelength 430nm, 30 ℃ of column temperatures, theoretical cam curve is not less than 4000 by the curcumin peak.
Said Radix Astragali extract is to take by weighing the Milkvetch Root coarse powder, the alcohol reflux with 70% 2 times; 12 times the dehydrated alcohol extraction 1.5h that adds for the first time Milkvetch Root weight adds 8 times dehydrated alcohol extraction 1h of Milkvetch Root weight, merge extractive liquid, the second time; Filter, decompression filtrate recycling ethanol is not to there being the alcohol flavor, and it is an amount of to add water; Leave standstill, filter, get Radix Astragali sample extracting solution.Get Radix Astragali ethanol extract (150mg/ml), reclaim ethanol and extremely do not have the alcohol flavor, cross macroporous adsorptive resins, elder generation is with the water elution of 4 times of resin volumes; Continue with 30% ethanol flush away impurity of 8 times of resin volumes, 70% ethanol elution of 8 times of resin volumes of reuse is collected 70% ethanol elution; Concentrating under reduced pressure ethanol, drying and crushing gets Radix Astragali extract; Yield is about 3%, and these article are flaxen powder, mildly bitter flavor; Spectrophotometry, these article contain total saponins and are no less than 75.0% in astragaloside; The HPLC method is measured, and the every 1g of these article contains the Radix Astragali in astragaloside (C41H68O14), must not be less than 7.2%.
Radix Astragali total saponins: Radix Astragali extract is measured with spectrophotography (appendix V of Chinese Pharmacopoeia version in 2005), and these article contain Radix Astragali total saponins and are no less than 75.0% in astragaloside.
Astragaloside: measure according to HPLC (an appendix VI of Chinese Pharmacopoeia version in 2005 D).Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filler; Mobile phase: acetonitrile-water (35: 65); Evaporative light scattering detector.Theoretical cam curve is not less than 4000 by the astragaloside chromatographic peak.
The present invention is a capsule, and content is faint yellow to the yellowish-brown granule, feeble QI, and mildly bitter flavor, strengthening spleen and lung is mended in the removing blood stasis with potent drugs circulation of qi promoting.Be used for the treatment of diabetes and early diabetes nephropathy, during treatment, oral the present invention, one time 2,3 times on the one.
Method for preparing of the present invention is simple, and compatibility is proper, and good effect has no side effect, and aspect treatment diabetes and the early diabetes nephropathy remarkable result is being arranged, and is the innovation on treatment diabetes and the early diabetes nephropathy, has huge economic and social benefit.
Four, the specific embodiment
Do detailed explanation below in conjunction with the practical situation specific embodiments of the invention.
Embodiment 1
The present invention is processed by Rhizoma Curcumae Longae extract 105g, Radix Astragali extract 65g, gets Rhizoma Curcumae Longae extract and Radix Astragali extract, pulverizes 100 mesh sieves, and mix homogeneously adds starch, each 9g mixing of Pulvis Talci, granulates, and incapsulates, and promptly gets every 0.3g.
Embodiment 2
The present invention also can be processed by Rhizoma Curcumae Longae extract 117g, Radix Astragali extract 75g, gets Rhizoma Curcumae Longae extract and Radix Astragali extract, pulverizes 100 mesh sieves, and mix homogeneously adds starch, each 11g mixing of Pulvis Talci, granulates, and incapsulates, and promptly gets every 0.3g.
Embodiment 3
The present invention also can be processed by Rhizoma Curcumae Longae extract 130g, Radix Astragali extract 85g, gets Rhizoma Curcumae Longae extract and Radix Astragali extract, pulverizes 100 mesh sieves, and mix homogeneously adds starch, each 8g mixing of Pulvis Talci, granulates, and incapsulates, and promptly gets every 0.3g.
The traditional Chinese medical science is through the unbalance negative and positive of adjustment; Improve self utilization and conversion capability to blood glucose; Thereby blood sugar lowering, and obstructed stimulation oversaturation B emiocytosis islets of langerhans blood sugar lowering usually use treatment by Chinese herbs to say from objective sense to have protection, the effect of reparation B cell.Though the Chinese medicine dynamics is not good enough aspect blood sugar control, curative effect is very remarkable aspect relieve clinical symptoms, and the rapid reduction of patient clinical symptoms of Chinese medicine can be brought the confidence of curing the disease, treating both the principal and secondary aspects of a disease to the patient.Western medicine to complication particularly diabetic nephropathy do not have special efficacious therapy method and medicine, and use treatment by Chinese herbs diabetic complication determined curative effect.Chinese medicine is the All Pure Nature medical material, and with the Western medicine contrast, as long as dialectical correct, toxic and side effects is minimum, and B cell is not had stimulation, the hypoglycemia incident can not take place; As long as compatibility is proper, the problem that did not exist lost efficacy in 5 years.It plays adjusting, strong effect to human body.
The present invention is effective to the treatment of diabetes and early diabetes nephropathy, and is proved fully that through experiment its experimental data is following:
Test the influence of a Rhizoma Zingiberis Recens astragalus capsules of the present invention to the causal diabetic mice
1 experiment material
1.1 medicine Rhizoma Zingiberis Recens astragalus capsules of the present invention, the preparation of pharmaceutical analysis subject; Metformin hydrochloride tablet, 0.25g/ sheet, 48/box, lot number 08102.
1.2 the reagent streptozotocin, sigma company; Glucose, Shantou Xilong Chemical Factory, Guangdong, lot number 081210; Citric acid (analytical pure), Hubei Province pharmaceuticals glass station, lot number 080523; Sodium citrate (analytical pure), chemical reagent wholesale department, Tianjin, lot number 080502; Normal saline, Zhengzhou chemistry pharmaceutcal corporation, Ltd, specification 500ml/ bottle, lot number 080809; Glucose assays test kit Zhe Jiangdong beats up biological company limited, lot number 081001.
1.3 instrument 752 spectrophotometers, Shanghai the 3rd analytical tool factory; The UV-2000 spectrophotometer, UNICO(Shanghai) Instruments Co., Ltd.; FA (N)/JA (N) series electronic balance, Shanghai Min Qiao precision instrument company limited; The TGL-16G High speed refrigerated centrifuge, Anting Scientific Instrument Factory, Shanghai; DZKW-4 type electronic thermostatic water-bath, the bright forever Medical Instruments in Beijing factory; Adjustable pipette, Shanghai Lei Bo Analytical Instrument Co., Ltd; The OLYMPUS optical microscope.
1.4 the animal mice, the Kunming kind, the SPF level, male, body weight: 17～20g; Medical animal experiment center, effluent south provides credit number: 410115.
1.5 the preparation of test solution preparation pH=4.2 citrate buffer: the citric acid 123ml of 0.1mol/L, 0.1mol/L sodium citrate 77ml faces and uses preceding preparation.The preparation of 10% formalin fixed liquid: 36% formalin 10ml is made into 100ml with distilled water.
2 experimental techniques and result
Get 72 of male mices; Evenly be divided into 6 groups at random, normally raised 3 days, behind the fasting 12h; Blank group tail vein injection is with the volume buffer; All the other 5 groups of tail vein injection streptozotocin 30mg/kg (0.02ml/10g, c:15mg/ml), and in the STZ of the 1st injection back filling in the 14th day isodose.Every morning is irritated clothes high concentration sucrose 30g/kg except that the blank group; Drink 5% sucrose water simultaneously, irritate respectively afternoon clothes large, medium and small dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention (600mg/kg, 300mg/kg, 150mg/kg, being made into concentration is 30mg/ml, 15mg/ml, 7.5mg/ml; Irritate the long-pending 0.2ml/10g of being of body of stomach); Metformin (0.5g/kg, being made into concentration is 25mg/ml, irritates the long-pending 0.2ml/10g of being of body of stomach) reaches with the volume normal saline.Administration every day 1 time is got blood in the 10th, 20 days tail veins of administration and is surveyed blood glucose value.The 30th day 1h after 12h last administration after the fasting, after fasting glucose was surveyed in the afterbody blood sampling, (5g/kg 0.4ml/10g), and respectively at irritating sugar back 30min, 60min, the blood sampling of 120min afterbody, surveyed the different time blood sugar level to irritate the clothes glucose.Get pancreas, kidney after putting to death mice, 10% formalin solution is fixed, and makes histopathology.The result sees table 1, table 2, table 3.Data analysis relatively adopts one factor analysis of variance with SPSS 12.0 for windows statistical softwares between the measurement data group, ranked data adopt Ridit to analyze.
Table 1 Rhizoma Zingiberis Recens astragalus capsules of the present invention to low dose of streptozotocin with sucrose
Cause the influence
of causal diabetic mice blood glucose
With the model group ratio, * * representes P＜0.01, and * representes P＜0.05
Can find out from table 1: with the blank control group ratio, each stage blood glucose value of model group significantly raises, and the modeling success is described.With the model group ratio, metformin can obviously reduce the 10th day blood glucose value (P＜0.05) after the administration of modeling mice, can significantly reduce the 20th, 30 day blood glucose value (P＜0.01); Heavy dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can obviously reduce the 20th day blood glucose value (P＜0.05), can significantly reduce the 30th day blood glucose value (P＜0.01); Middle dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention can significantly reduce the 30th day blood glucose value (P＜0.01), and the trend that reduces the 20th day blood glucose value is arranged; Low dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can significantly reduce the 30th day blood glucose value (P＜0.01) of mice, and the 10th, 20 day blood glucose value is not had obvious influence.Explain that Rhizoma Zingiberis Recens astragalus capsules of the present invention and metformin all can resist the blood sugar increasing of model mice to some extent.
Table 2 Rhizoma Zingiberis Recens astragalus capsules of the present invention causes with sucrose low dose of streptozotocin
The influence of causal diabetic mice carbohydrate tolerance
With the model group ratio, * * representes P＜0.01, and * representes P＜0.05
Can find out from last table: with the blank control group ratio, model group 60min, 120min blood sugar level after irritating sucrose solution all significantly increase, and explain that modeling mice carbohydrate tolerance reduces.With the model group ratio, metformin can obviously reduce the modeling mice and irritate 30min blood glucose value (P＜0.05) behind the sucrose solution, can significantly reduce and irritate sucrose solution 60min, 120min blood glucose value (P＜0.01); Heavy dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can obviously reduce 30min, 60min, 120min blood glucose value (P＜0.05); Middle dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention can obviously reduce 30min, 60min, 120min blood glucose value (P＜0.05); Low dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention obviously reduces 60min blood glucose value (P＜0.05).Explain that metformin, big or middle dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention can improve the carbohydrate tolerance reduction of diabetic mice.
Table 3 Rhizoma Zingiberis Recens astragalus capsules of the present invention is to the influence of diabetic mice kidney and pancreatic tissue
"-" is that matter was all normal between renal cells and glomerule reached; "+" renal cells has Mild edema; " ++ " renal cells extensive edema, a matter is with cell infiltration; " +++" renal cells extensive edema and a large amount of cell infiltration of a matter and messangial cell hypertrophy, capsular space narrows down.
"-" all enriches full for the main β cell cytosol of islet cells; "+" is that 2～4 cellular atrophies appear in the main β cell of islet cells, and cell cytosol obviously reduces; " ++ " is that 5～9 cellular atrophies appear in the main β cell of islet cells, and cell cytosol obviously reduces; " +++" be that the cellular atrophy more than 10 appears in islet cells β cell, endochylema obviously reduces.
Can find out from table 3: matter was normal basically between blank control group glomerule, renal cells reached.The cell infiltration in various degree of a most of renal cells edema of model group kidney and a matter through Ridit check P＜0.01, explains that kidney injury appears in the modeling mice.With the model group ratio, matter was normal between a part of glomerule, renal cells reached in the metformin group, the cell infiltration in various degree of a most of renal cells edema and a matter (P＞0.05); Dosage big or middle Rhizoma Zingiberis Recens astragalus capsules of the present invention group glomerule, renal cells and between matter normal (P＜0.01) basically; Edema in various degree appears in low dose of Rhizoma Zingiberis Recens astragalus capsules group renal cells of the present invention, and hyperplasia appears in glomerule, renal capsule narrow down (P＞0.05).Results suggest: middle dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention has protective effect to low dose of streptozotocin with the pathological changes that sucrose causes the kidney of causal diabetic mice, and effect is better than metformin.
Also can find out from table 3: the β cell cytosol each islet cells of blank control group animal enriches full, the obvious atrophy of β cell in the model group animal insulin cell, and endochylema obviously reduces, and through Ridit check P＜0.01, explains that injury of pancreas appears in the modeling mice.With the model group ratio, metformin can obviously be resisted the atrophy of part β cell in the islet cells, makes part endochylema improve (P＜0.05); Heavy dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can resist part β cellular atrophy, makes most of endochylema abundant (P＜0.05); In the atrophy that can resist most of β cell of dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention, make most of endochylema be improved significantly (P＜0.05); Low dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention reduces do not make significant difference (P＞0.05) to β cellular atrophy and the endochylema that modeling causes.Results suggest: big or middle dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention group and metformin group have protective effect to the islets of langerhans damage due to the streptozotocin.
Experiment two, Rhizoma Zingiberis Recens astragalus capsules of the present invention are to the influence of diabetic nephropathy model rat
1 experiment material
1.1 medicine Rhizoma Zingiberis Recens astragalus capsules of the present invention, the preparation of pharmaceutical analysis subject; Metformin hydrochloride tablet, 0.25g/ sheet, 48/box, lot number 08102.
1.2 the reagent streptozotocin, sigma company; Glucose, Shantou Xilong Chemical Factory, Guangdong, lot number 081210; Citric acid (analytical pure), Hubei Province pharmaceuticals glass station, lot number 080523; Sodium citrate (analytical pure), chemical reagent wholesale department, Tianjin, lot number 080502; Normal saline, Zhengzhou chemistry pharmaceutcal corporation, Ltd, specification 500ml/ bottle, lot number 080809; Glucose assays test kit Zhe Jiangdong beats up biological company limited, lot number 081001; MAU detection kit, Nanjing are built up bio-engineering research institute, lot number 081120; Scr detection kit, Nanjing are built up bio-engineering research institute, lot number 081114; BUN detection kit, Nanjing are built up bio-engineering research institute, lot number 081108; NO detection kit, Nanjing are built up bio-engineering research institute, lot number 081110; NOS detection kit, Nanjing are built up bio-engineering research institute, lot number 081116
1.3 instrument 752 spectrophotometers, Shanghai the 3rd analytical tool factory; The UV-2000 spectrophotometer, UNICO(Shanghai) Instruments Co., Ltd.; FA (N)/JA (N) series electronic balance, Shanghai Min Qiao precision instrument company limited; The TGL-16G High speed refrigerated centrifuge, Anting Scientific Instrument Factory, Shanghai; DZKW-4 type electronic thermostatic water-bath, the bright forever Medical Instruments in Beijing factory; Adjustable pipette, Shanghai Lei Bo Analytical Instrument Co., Ltd; The OLYMPUS optical microscope; The H-7500 of Hitachi transmission electron microscope.
1.4 animal rat, the SPF level, Wistar, male, body weight: 165～185g provides credit number by medical animal experiment center, Henan: 410117.
1.5 the preparation of test solution preparation pH=4.2 citrate buffer: the citric acid 123ml of 0.1mol/L, 0.1mol/L sodium citrate 77ml faces and uses preceding preparation.The preparation of 10% formalin fixed liquid: 36% formalin 10ml is made into 100ml with distilled water.The preparation of 4% glutaraldehyde fixative: sodium hydrogen phosphate 35.61g is made into 1000ml with distilled water; Sodium dihydrogen phosphate 27.60g is made into 1000ml with distilled water.Add 25% glutaraldehyde 16.0ml after getting disodium phosphate soln 40.5ml and sodium dihydrogen phosphate 9.5ml mixing, add distilled water again to 100ml.
2 experimental techniques and result
Get 100 of rats, tail vein injection streptozotocin 60mg/kg behind the fasting 12h (use pH is 4.2 citrate buffer preparation) selects 10 else and is made as complete blank group, the isopyknic citrate buffer of tail vein injection.The 10th day afterbody got blood and surveyed blood glucose value, and 50 of the rats of choosing blood glucose value＞11.1mmol/L and having obvious polydipsia, polyphagia, polyuria symptom are divided into 5 groups at random by blood glucose value; Irritate big-and-middle, little, dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention (300mg/kg, 150mg/kg, 75mg/kg, being made into concentration is 30mg/ml, 15mg/ml, 7.5mg/ml, irritates the long-pending 1ml/100g that is of body of stomach) respectively; Diformin tablet 250mg/kg (25mg/ml, 1ml/100g), model group and blank group give with the volume normal saline; Administration every day 1 time, successive administration 40 days is respectively at the 10th, 20,30 day survey fasting blood sugar of administration; Each preceding urine amount of observing of blood glucose of surveying, in last survey blood glucose fasting the previous day, metabolic cage is collected 12 hours nocturia; Centrifugal (3000rpm/min) 10 minutes, cold preservation substitutes; Behind the perfusion 2h, get blood, separation of serum.Put to death and to get pancreas behind the rat, kidney is made routine pathology, get kidney and make transmission electron microscope observing.Data analysis relatively adopts one factor analysis of variance with SPSS 10.0 for windows statistical softwares between the measurement data group, ranked data adopt Ri dit to analyze.The assay method of thymic cortex thickness and lymphocyte number: record thymic cortex the widest part and the narrowest place with micrometer, averaging is cortical thickness; The micrometer baseline is pressed in the LC on the thymic cortex, and averaging is lymphocyte number.The result sees table 4, table 5, table 6, table 7, table 8.
Table 4 Rhizoma Zingiberis Recens astragalus capsules of the present invention is to the influence
of diabetic nephropathy model rat blood sugar
With the model group ratio, * * representes P＜0.01, and * representes P＜0.05
Can find out from last table: with the blank control group ratio, the model group blood sugar level is significantly higher than blank control group (P＜0.01), and it is successful to explain that streptozotocin is made diabetic nephropathy model.With the model group ratio, metformin can significantly reduce the 20th, 30,40 day blood glucose value (P＜0.01) after the administration, and the 10th day blood glucose value do not had obvious influence; Heavy dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can obviously reduce the 20th day blood glucose value (P＜0.05), can significantly reduce the 30th, 40 day blood glucose value (P＜0.01); Middle dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention can significantly reduce the 20th, 30,40 day blood glucose value, and the 10th day blood glucose value trend of reduction is arranged; Low dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can obviously reduce the 30th day blood glucose value (P＜0.05), significantly reduce the 40th day blood glucose value (P＜0.05).
Table 5 Stigma Maydis total saponins is to the influence
of diabetic nephropathy model rat serum creatinine, UCr, renal index
With the model group ratio, * * representes P＜0.01, and * representes P＜0.05
Can find out from last table: model group rat serum creatinine, UCr, renal index significantly raise (P＜0.01).With the model group ratio, large, medium and small dosage Stigma Maydis total saponins can significantly reduce serum creatinine, UCr, renal index value (P＜0.01).
Table 6 Rhizoma Zingiberis Recens astragalus capsules of the present invention is to the influence
of diabetic nephropathy model rat serum BUN, MAU, Ccr
Compare with model group, * * representes P＜0.01, and * representes P＜0.05
Can find out that from last table with the blank control group ratio, model group BUN, MAU, Ccr significantly raise.With the model group ratio, metformin can significantly reduce BUN, MAU value (P＜0.01); Dosage big or middle Rhizoma Zingiberis Recens astragalus capsules of the present invention can significantly reduce BUN, MAU value (P＜0.01); Low dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can obviously reduce BUN (P＜0.05), significantly reduce MAU value (P＜0.01), in, low dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can obviously reduce Ccr (P＜0.05).
Table 7 Rhizoma Zingiberis Recens astragalus capsules is to the influence
of diabetic nephropathy model rat NO, NOS level
Compare with model group, * * representes P＜0.01, and * representes P＜0.05;
Can find out from last table: with the blank control group ratio, model group NO, NOS be significantly rising (P＜0.01) all.With the model group ratio, metformin can significantly reduce NO, NOS level (P＜0.01); Large, medium and small dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention can significantly reduce NO, NOS level (P＜0.01).
Table 8 Rhizoma Zingiberis Recens astragalus capsules of the present invention is to the influence of diabetic nephropathy model rat kidney and pancreatic tissue
"-" is that matter was all normal between renal cells and glomerule reached; "+" renal cells has Mild edema; " ++ " renal cells extensive edema, a matter is with cell infiltration; " +++" renal cells extensive edema and a large amount of cell infiltration of a matter and messangial cell hypertrophy, capsular space narrows down.
"-" all enriches full for the main β cell cytosol of islet cells; "+" is that 2～4 cellular atrophies appear in the main β cell of islet cells, and cell cytosol obviously reduces; " ++ " is that 5～9 cellular atrophies appear in the main β cell of islet cells, and cell cytosol obviously reduces; " +++" be that the cellular atrophy more than 10 appears in islet cells β cell, endochylema obviously reduces.
Can find out from last table: matter was normal basically between the glomerule of blank control group rat, renal cells reached.The model group rat kidney is renal cells edema and matter cell infiltration in various degree widely, through Ridit check P＜0.01, explains that the kidney of modeling rat damages.With the model group ratio; Heavy dose of and middle dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention can be protected glomerule, renal cells; Make it and significantly improve (P＜0.01), metformin group and low dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can partly improve the extensive edema of renal cells and the infiltration (P＜0.05) of a matter inflammatory cell.Results suggest: Rhizoma Zingiberis Recens astragalus capsules group of the present invention has the better protect effect to the kidney injury of rat model due to the streptozotocin.
Also can find out from last table: the β cell cytosol each islet cells of blank control group rat enriches full, the obvious atrophy of β cell in the model group Islet cells, and endochylema obviously reduces, and through Ridit check P＜0.01, explains that the modeling pancreas in rat damages.With the model group ratio, metformin can partly be resisted the β cellular atrophy, makes part cell cytosol be restored (P＞0.05); Dosage big or middle Rhizoma Zingiberis Recens astragalus capsules of the present invention can resist most of β cellular atrophy, makes most of endochylema significantly improve (P＜0.01); Low dose of Rhizoma Zingiberis Recens astragalus capsules of the present invention can resist part β cellular atrophy, makes part endochylema make moderate progress (P＞0.05).Results suggest: big or middle dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention has protective effect to the islets of langerhans damage due to the streptozotocin, and effect obviously is better than the metformin group.
Table 9 Rhizoma Zingiberis Recens astragalus capsules of the present invention is to the influence of diabetic nephropathy model rat chest gland cortical thickness, lymphocyte number
Compare with model group, * * representes P＜0.01, and * representes P＜0.05
Under last table and light microscopic, can find out: pathological changes such as thymic cortex appearance cortical thickness attenuation, the cell in various degree of model group rat is sparse the observation of diabetes rat thymic tissue.With the model group ratio, large, medium and small dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention group rat chest gland cortical thickness is significantly improved (P＜0.01), and lymphocyte number has remarkable increase (P＜0.01).
Table 10 Rhizoma Zingiberis Recens astragalus capsules of the present invention is to diabetic nephropathy model rat renal tubular epithelial cells mitochondrion
Influence (the unit: μ m of body density (Vv), specific surface (δ), membrane density (δ m)
3, μ m
2)
Compare with model group, * * representes P＜0.01, and * representes P＜0.05
Table 11 Rhizoma Zingiberis Recens astragalus capsules of the present invention is to the diabetic nephropathy model rat renal tubular epithelial cells
Influence (the unit: μ m of nuclear specific surface (δ), different, euchromatic body density (Vv)
3, μ m
2)
Compare with model group, * * representes P＜0.01, and * representes P＜0.05
Can find out from last table: the blank control group proximal tubular epithelial cells has marshalling and dense microvilli, has abundant mitochondria to have most of mitochondrial crista dense arrangement in the endochylema; The few part broadening in ridge gap, the Distal convoluted tubule epithelial cell does not have microvillus, and endochylema contains abundant mitochondria; Part mitochondrial crista dense arrangement, part mitochondrial crista arrangement disorder, chamber broadening to some extent between ridge; Nuclear goes out many ghosts week, presents the edema state; The model group proximal tubular epithelial cells has marshalling and dense microvilli is the cross section; Nucleus is irregular shrinkage; Chromatin presents that heterochromatin increases and euchromatin reduces, and the mitochondrion great majority are the edema state in the endochylema, and it is irregular that ridge appears; The chamber differs in size between ridge, dissolving of part mitochondrial crista or disappearance.The Distal convoluted tubule epithelial cell presents the edema state, particularly cavity not of uniform size, stromatolysis occur around the nuclear in the endochylema; Mitochondrion obviously reduces, and ridge is partly dissolved disappearance, and the chamber increases between ridge; Few part mitochondrial crista marshalling is intensive, and fold partly appears in nuclear membrane, the chromatin degraded.With the model group ratio, each dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention can improve the change of above-mentioned ultrastructure in kidney to some extent.Data t inspection show in groups; Modeling metanephric tubule epithelial cell mitochondrion body density (mitochondrion Vv), membrane density (δ m), euchromatin body density (euchromatin Vv) all significantly reduce (P＜0.01), specific surface (δ), nuclear specific surface (nuclear δ), all significantly risings (P＜0.01) of heterochromatin body density (heterochromatin Vv).Large, medium and small Rhizoma Zingiberis Recens astragalus capsules of the present invention all can significantly raise renal cells mitochondrion Vv, δ m, euchromatin Vv (P＜0.01) all can significantly reduce δ, nuclear δ, heterochromatin Vv (P＜0.01).High proximal convoluted tubule nuclear (δ), heterochromatin Vv (P＜0.01) can significantly fall in the large, medium and small dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention renal tubular epithelial proximal convoluted tubule euchromatin Vv (P＜0.01) that all can significantly raise; Large, medium and small dosage Rhizoma Zingiberis Recens astragalus capsules of the present invention all can significantly reduce renal tubular epithelial Distal convoluted tubule nuclear (δ), euchromatin Vv (P＜0.01), and Distal convoluted tubule heterochromatin Vv (P＜0.01) can significantly raise.
Conclusion: Rhizoma Zingiberis Recens astragalus capsules of the present invention can obviously reduce and improve low dose of streptozotocin and increase sugared diet induced hyperglycemia mice model blood sugar level and carbohydrate tolerance reduction; Can obviously resist β cellular atrophy in the mouse islets cell; Endochylema is obviously recovered; Can improve modeling mice pancreatic histopathology to a certain extent and change, the kidney of diabetic mice is had protective effect, act on suitable with metformin; Various dose Rhizoma Zingiberis Recens astragalus capsules of the present invention can reduce due to the streptozotocin blood glucose, creatinine, blood urea nitrogen, NO, NOS in the diabetes model rat blood serum in various degree; The level of indexs such as microdose urine protein, creatinine and endogenous creatinine clearance rate in the urine; Show through the om observation analysis: Rhizoma Zingiberis Recens astragalus capsules of the present invention can be protected the islet cells of modeling rat, and antagonism modeling medicine is to the damage of islet cells; Kidney to the modeling rat has protective effect, can alleviate Distal convoluted tubule epithelial cell edema state; The electron microscopic observation analysis shows: Rhizoma Zingiberis Recens astragalus capsules of the present invention can increase rat renal tubular epithelial cells mitochondrion quantity, reduces mitochondrion heterochromatin quantity, the dissolving and the disappearance that alleviate mitochondrial crista; Through the improvement of tissue microstructure being helped delaying to a certain extent the pathological process of diabetes rat kidney.
Analysis through experimental result; Rhizoma Zingiberis Recens astragalus capsules of the present invention delay and improve the effect of nephropathy due to the diabetes rat maybe be with its blood sugar lowering, reduce microdose urine protein, creatinine, blood urea nitrogen, endogenous creatinine clearance rate level, reduce serum NO, NOS level; The factors such as histo pathological change of improving the modeling animal are relevant, thereby delay the generation and the development of diabetic nephropathy.
Claims (4)
1. a Rhizoma Zingiberis Recens astragalus capsules of treating diabetes and early diabetes nephropathy is characterized in that, is processed by Rhizoma Curcumae Longae extract 105～130g, Radix Astragali extract 65～85g, starch, Pulvis Talci; Rhizoma Curcumae Longae extract and Radix Astragali extract were pulverized 100 mesh sieves; Mix homogeneously adds starch, Pulvis Talci is mixed again, granulates; Incapsulate, promptly get;
Starch, Pulvis Talci addition≤Rhizoma Curcumae Longae extract, Radix Astragali extract, starch, Pulvis Talci gross weight and 10%, starch, Pulvis Talci weight ratio are 1: 1;
Said Rhizoma Curcumae Longae extract is, takes by weighing the Rhizoma Curcumae Longae medical material after the pulverizing, puts into microwave extraction tank; Amount of ethanol is 20 times of Rhizoma Curcumae Longae medical material weight, and promptly Rhizoma Curcumae Longae and alcoholic acid weight ratio are 1: 20, extracts 50 ℃ of temperature; Extraction time 30min extracts power 200W, and it is subsequent use to get the Rhizoma Curcumae Longae crude extract;
The S-8 purification with macroreticular resin: the S-8 macroporous adsorbent resin is earlier with soak with ethanol 24 hours, is washed with distilled water to that not have alcohol afterwards subsequent use; With the macroporous adsorbent resin wet method dress post of handling well; The Rhizoma Curcumae Longae crude extract is adsorbed through chromatographic column with the 1mL/min flow velocity, and elder generation is with the water elution impurity of 4 times of resin volumes, the dehydrated alcohol eluting of 8 times of resin volumes of reuse; Collect the dehydrated alcohol eluent; Concentrating under reduced pressure ethanol, drying and crushing gets Rhizoma Curcumae Longae extract; Spectrophotometry, these article contain total curcumin and are no less than 58.0% in curcumin; The HPLC method is measured, and the every 1g of these article contains Rhizoma Curcumae Longae in curcumin (C21H20O 6), must not be less than 15.0%;
Said Radix Astragali extract is to take by weighing the Milkvetch Root coarse powder, the alcohol reflux with 70% 2 times; 12 times the dehydrated alcohol extraction 1.5h that adds for the first time Milkvetch Root weight adds 8 times dehydrated alcohol extraction 1h of Milkvetch Root weight, merge extractive liquid, the second time; Filter, decompression filtrate recycling ethanol is not to there being the alcohol flavor, and it is an amount of to add water; Leave standstill, filter, get Radix Astragali sample extracting solution; Get Radix Astragali ethanol extract, 150mg/ml reclaims ethanol to there not being the alcohol flavor; Cross macroporous adsorptive resins, with the water elution of 4 times of resin volumes, continue earlier with 30% ethanol flush away impurity of 8 times of resin volumes; 70% ethanol elution of 8 times of resin volumes of reuse is collected 70% ethanol elution, concentrating under reduced pressure ethanol; Drying and crushing gets Radix Astragali extract; Spectrophotometry, these article contain total saponins and are no less than 75.0% in astragaloside; The HPLC method is measured, and the every 1g of these article contains the Radix Astragali in astragaloside (C41H68O14), must not be less than 7.2%.
2. the Rhizoma Zingiberis Recens astragalus capsules of treatment diabetes according to claim 1 and early diabetes nephropathy is characterized in that, is processed by Rhizoma Curcumae Longae extract 105g, Radix Astragali extract 65g; Get Rhizoma Curcumae Longae extract and Radix Astragali extract, pulverized 100 mesh sieves, mix homogeneously; Add starch, each 9g mixing of Pulvis Talci, granulate, incapsulate; Promptly get every 0.3g.
3. the Rhizoma Zingiberis Recens astragalus capsules of treatment diabetes according to claim 1 and early diabetes nephropathy is characterized in that, is processed by Rhizoma Curcumae Longae extract 117g, Radix Astragali extract 75g; Get Rhizoma Curcumae Longae extract and Radix Astragali extract, pulverized 100 mesh sieves, mix homogeneously; Add starch, each 11g mixing of Pulvis Talci, granulate, incapsulate; Promptly get every 0.3g.
4. the Rhizoma Zingiberis Recens astragalus capsules of treatment diabetes according to claim 1 and early diabetes nephropathy is characterized in that, is processed by Rhizoma Curcumae Longae extract 130g, Radix Astragali extract 85g; Get Rhizoma Curcumae Longae extract and Radix Astragali extract, pulverized 100 mesh sieves, mix homogeneously; Add starch, each 8g mixing of Pulvis Talci, granulate, incapsulate; Promptly get every 0.3g.
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| 余俊先等.黄茂甲昔防治糖尿病大鼠周围神经病变的研究.《2008年中国药理学会药学发展前沿论坛》.2008,101. * |
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