CN110787263A

CN110787263A - Traditional Chinese medicine composition for treating diabetic nephropathy and application thereof

Info

Publication number: CN110787263A
Application number: CN201911248582.4A
Authority: CN
Inventors: 许鑫; 王恒斌; 潘耀宗; 李全; 翟云良; 罗年翠; 张邦国; 张丹丹; 戴晶晶
Original assignee: Leiyunshang Pharmaceutical Group Co Ltd
Current assignee: Leiyunshang Pharmaceutical Group Co Ltd
Priority date: 2019-12-09
Filing date: 2019-12-09
Publication date: 2020-02-14

Abstract

The invention discloses an application of a traditional Chinese medicine composition in preparing a medicine for treating diabetes and diabetic nephropathy. Experimental research shows that the traditional Chinese medicine composition preparation has a good effect of reducing fasting plasma glucose of diabetic nephropathy rats, and can remarkably reduce the contents of serum creatinine, urea nitrogen, kidney coefficient and urine protein of the diabetic nephropathy rats. Meanwhile, the composition has good effect of preventing and treating the fibrosis of the renal tubular epithelial cells. Can be used for developing new pharmaceutical preparations for clinical indications.

Description

Traditional Chinese medicine composition for treating diabetic nephropathy and application thereof

Technical Field

The invention belongs to the field of traditional Chinese medicine pharmacy, and particularly relates to a traditional Chinese medicine composition for treating diabetic nephropathy and application thereof.

Background

Diabetes is a common and frequently occurring disease worldwide and has become the third largest disease after tumors and cardiovascular diseases. Diabetes is clinically often associated with other diseases, such as diabetes associated with nephropathy and diabetes associated with hyperlipidemia. On one hand, diabetes complicated with nephropathy is caused by yin deficiency and dryness-heat, kidney qi deficiency and loss of essence; on the other hand, blood stasis is formed by burning blood, blood stasis blocks kidney collaterals, dysfunction of opening and closing and fine and slight diarrhea can all cause the diabetic kidney damage, and the life quality of people is seriously influenced.

At present, most of the medicines for treating diabetes are used, although western medicines used clinically have a certain curative effect, most of the western medicines have large adverse reactions, such as rebound after stopping taking the medicines, liver and kidney damage of patients, rhabdomyolysis and other side effects, which can treat symptoms and not root causes, while the traditional Chinese medicine has unique advantages in the aspect of treating diabetes.

A Chinese medicinal capsule, LISHISHAN Capsule, is prepared by LIYI pharmaceutical industry, and can be used for treating chronic primary nephritis with mild or moderate degree, with the Chinese medicinal standard Z20020086. No treatment for diabetes or diabetic nephropathy has been reported.

Disclosure of Invention

The purpose of the invention is as follows: the invention aims to provide a traditional Chinese medicine composition for treating diabetic nephropathy and application thereof.

The technical scheme is as follows: the purpose of the invention is realized by the following technical scheme:

the application of a traditional Chinese medicine composition in preparing a medicament for treating diabetes comprises radix astragali, fructus Lycii, rhizoma Dioscoreae, Coicis semen, radix scrophulariae, radix Glehniae, Ecliptae herba, placenta hominis, Eucommiae cortex, Notoginseng radix, herba Leonuri, Hirudo, periostracum Cicadae, semen plantaginis, and Achyranthis radix.

The application of a traditional Chinese medicine composition in preparing a medicine for treating diabetic nephropathy comprises radix astragali, fructus Lycii, rhizoma Dioscoreae, Coicis semen, radix scrophulariae, radix Glehniae, Ecliptae herba, placenta hominis, Eucommiae cortex, Notoginseng radix, herba Leonuri, Hirudo, periostracum Cicadae, semen plantaginis, and Achyranthis radix.

Preferably, for the above-mentioned use, said diabetes is type II diabetes.

Preferably, the above-mentioned application is to prepare the extract of the traditional Chinese medicine composition and a pharmaceutically acceptable carrier into capsules, tablets, granules, pills or oral liquid.

Preferably, the extract of the Chinese medicinal composition is prepared by decocting the Chinese medicinal materials in water or extracting with ethanol under reflux, continuously refluxing, ultrasonically extracting, and concentrating.

Preferably, the traditional Chinese medicine composition is prepared from 680-1360 parts of astragalus membranaceus, 170-340 parts of wolfberry fruits, 255-510 parts of Chinese yams, 170-340 parts of coix seeds, 170-340 parts of radix scrophulariae, 170-340 parts of radix glehniae, 170-340 parts of eclipta, 38-76 parts of human placenta, 340-680 parts of eucommia ulmoides, 153-306 parts of pseudo-ginseng, 510-1020 parts of motherwort, 38-76 parts of leeches, 51-102 parts of cicada slough, 170-340 parts of plantain seeds and 170-340 parts of achyranthes bidentata.

Preferably, the traditional Chinese medicine composition is prepared from 680 parts of astragalus mongholicus, 170 parts of wolfberry, 255 parts of Chinese yam, 170 parts of coix seeds, 170 parts of radix scrophulariae, 170 parts of radix glehniae, 170 parts of eclipta, 38 parts of human placenta, 340 parts of eucommia ulmoides, 153 parts of pseudo-ginseng, 510 parts of motherwort, 38 parts of leech, 51 parts of cicada slough, 170 parts of semen plantaginis and 170 parts of radix achyranthis bidentatae.

Preferably, the preparation method of the traditional Chinese medicine composition comprises the following steps:

(1) pulverizing Notoginseng radix, placenta hominis and Hirudo into fine powder, sieving with 100 mesh sieve, and sterilizing;

(2) decocting radix astragali, fructus Lycii, rhizoma Dioscoreae, Coicis semen, radix scrophulariae, radix Glehniae, Ecliptae herba, Eucommiae cortex, herba Leonuri, periostracum Cicadae, semen plantaginis, and Achyranthis radix with water twice (1 hr each time), mixing decoctions, filtering, concentrating the filtrate under reduced pressure to obtain fluid extract, adding ethanol, stirring, standing for 24 hr, collecting supernatant, recovering ethanol under reduced pressure, and concentrating to obtain fluid extract; adding the fine powder of the pseudo-ginseng, the human placenta and the leech in the step (1), uniformly mixing, granulating, drying, crushing into fine powder, adding a pharmaceutically acceptable carrier, and preparing into a pharmaceutical preparation.

(1) firstly, taking pseudo-ginseng, human placenta and leech according to a weight ratio, crushing the pseudo-ginseng, the human placenta and the leech into fine powder, sieving the fine powder by a 100-mesh sieve, and sterilizing the fine powder for later use;

(2) then, decocting the astragalus, the wolfberry fruit, the Chinese yam, the coix seed, the figwort root, the radix glehniae, the yerbadetajo herb, the eucommia bark, the motherwort herb, the cicada slough, the plantain seed and the achyranthes root twice by weight in water for 1 hour each time, combining the decoction, filtering, concentrating the filtrate under reduced pressure to obtain clear paste with the relative density of 1.16-1.20 at 60 ℃, adding ethanol to ensure that the alcohol content reaches 63 percent, stirring uniformly, standing for 24 hours, taking supernatant, recovering the ethanol under reduced pressure, and concentrating to obtain clear paste with the relative honey degree of 1.36-1.39 at 60 ℃; adding the fine powder of the pseudo-ginseng, the human placenta and the leech in the step (1), uniformly mixing, granulating, drying, crushing into fine powder, and adding a pharmaceutically acceptable carrier to prepare the pharmaceutical preparation.

Pharmacological experiment results show that the traditional Chinese medicine composition has obvious effects of preventing and treating diabetes and diabetic nephropathy.

Has the advantages that: the traditional Chinese medicine composition preparation for treating diabetic nephropathy provided by the invention has the following advantages:

experimental results show that the traditional Chinese medicine composition preparation for treating diabetic nephropathy has good effect of reducing fasting plasma glucose of diabetic nephropathy rats, and can remarkably reduce the contents of serum creatinine, urea nitrogen, kidney coefficient and urine protein of diabetic nephropathy rats. And the experimental result shows that the invention also has good effect of preventing and treating the fibrosis of the renal tubular epithelial cells. In the experimental process, no obvious toxic or side effect is seen, and the safety is good.

Detailed Description

The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the specific material ratios and results thereof described in the examples are merely illustrative of the invention and should not limit the invention as detailed in the claims. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Example 1

A traditional Chinese medicine composition for treating diabetic nephropathy is prepared by the following method:

(1) 153g of pseudo-ginseng, 38g of human placenta and 38g of leech are taken according to the weight proportion and crushed into fine powder, the fine powder is sieved by a 100-mesh sieve, and the fine powder is sterilized for standby;

(2) 680g of astragalus mongholicus, 170g of wolfberry fruit, 255g of Chinese yam, 170g of coix seed, 170g of radix scrophulariae, 170g of radix glehniae, 170g of eclipta, 340g of eucommia ulmoides, 510g of motherwort, 51g of cicada slough, 170g of semen plantaginis and 170g of radix achyranthis bidentatae are taken according to the weight ratio, water is added for decoction twice, 1 hour is carried out each time, decoction liquid is combined, filtration is carried out, filtrate is decompressed and concentrated to clear paste with the relative density of 1.16-1.20 at the temperature of 60 ℃, ethanol is added to ensure that the ethanol content reaches 63 percent, stirring is carried out, standing is carried out for 24 hours, supernatant is taken, decompressed, ethanol is recovered, and concentrated; adding the fine powder of the pseudo-ginseng, the human placenta and the leech in the step (1), uniformly mixing, granulating, drying, crushing into fine powder, drying, granulating, sieving, coating, and encapsulating to prepare 1000 granules.

Example 2 experiments on lowering blood sugar, lowering blood lipid and protecting kidney injury

The method comprises the following steps of taking 14-16-week-old SD rats with body weight (220 +/-20) G and male barrier environment rats (SPF grade), adaptively feeding the SD rats for 5 days, feeding 10 rats in a blank group with conventional feed during the experiment, feeding the rest rats with high glucose and high fat and 0.5% glucose water, injecting STZ into abdominal cavities for 3 consecutive days at 30mg/kg after 3 weeks, continuously feeding the rats with high glucose and high fat and glucose water for 4 days, fasting and keeping water, taking blood from tail veins after 12 hours to measure fasting blood glucose, and successfully modeling by taking blood glucose value higher than 13.1mmol/L, dividing the successfully-modeled diabetic rats into 5 groups according to fasting blood glucose level, wherein 10 rats in each group are respectively a model group and a stomach metformin (positive control drug) group (13.5 mg/kg. d) after 15 days, taking capsules of a high-dose group (1.25G/kg. d), a medium-dose group (0.65G/kg. d), a low-dose group (0.25G/kg. d), taking capsules of the traditional Chinese medicinal composition prepared in the example 1, a medium-dose group (1.25G/kg. d), a medium-dose group, a medium-dose of rat), a medium-dose of rat, a rat serum albumin (0.5G/kg), a rat with a serum glucose concentration serum, a serum of a rat with a serum of a rat with a serum, a serum of a serum, a rat with a serum.

1. Effects on fasting blood glucose

The fasting blood glucose of rats in the blank group, the model group, the positive drug group and the traditional Chinese medicine composition capsule administration group is measured by tail vein blood sampling, and the result shows that the fasting blood glucose of rats in the positive drug administration group and the traditional Chinese medicine composition capsule administration group is obviously lower than that of the model group (p is less than 0.01). Specific results are shown in table 1.

TABLE 1 Effect of drugs on fasting plasma glucose in diabetic model rats

Note: # was significantly different from the normal group. Significant difference P <0.05 compared to model group; very significant difference P <0.001 compared to model group.

2. Influence on renal function injury-related index

The serum creatinine and urea nitrogen contents in the rat serum of the blank group, the model group and the Chinese medicinal composition capsule group are measured by a kit, the protein content in the urine is detected for 24h by a Coomassie brilliant blue G-250 method, and the kidney index (the kidney index is (left kidney mass + right kidney mass)/(rat body mass x 100)) is calculated. The results show that the contents of serum creatinine, urea nitrogen, 24h urine protein and the kidney index value of the rat in the capsule group of the traditional Chinese medicine composition are obviously lower than those in the model group (p is less than 0.05). As shown in table 2.

TABLE 2 Effect of drugs on renal function-related indices in diabetic rats

3. Influence on the pathological state of the kidney tissue

After administration, taking kidney tissues of rats of each group to perform HE (human immunodeficiency syndrome) dyeing, PAS (PAS) dyeing and Masson dyeing, wherein renal tubules and glomeruli of the rats of a normal group seen under a light microscope are normal, renal tubular epithelium vacuolization, renal interstitial mass inflammatory cell infiltration and glomerular cyst dilatation of a model group are normal, and the kidney structure of the rats of a capsule administration group of the traditional Chinese medicine composition is close to that of the normal group.

Example 3 functional experiment for improving renal tubule fibrosis

1. Test cell

The tubular epithelial cell line HK-2 was cultured in a DMEM high-glucose medium containing 10% fetal bovine serum, 100U/mL penicillin and 100. mu.g/mL streptomycin, and placed at 37 ℃ in a medium containing 5% CO₂An incubator.

2. Medicine

Positive drug DPI (NADPH oxidase inhibitor), purchased from Calbiochem Corp. (La Jolla, CA).

3. Grouping and processing of cells

Plating cells with 96-well plate, pretreating with DMEM high-sugar medium, 37 deg.C, 5% CO₂Culturing for 6h to make it in synchronization state. The solution was changed every 24 h. The experiment is divided into: blank control, i.e. no intervention factor added); a high sugar treatment group; the administration groups were DPI (10. mu.M) and the capsule treatment group (50. mu.M) of the Chinese medicinal composition of example 1, respectively.

4. Analysis of experiments

The relative contents of E-cadherin, α -SMA, FN, p-ERK, NOX1, NOX2, NOX4, TGF- β - β RI, TGF- β RII, Smad2, Smad3 and Smad7 are determined by a Western blot analysis method.

5. Results of the experiment

(1) Effect on high sugar-induced HK-2 cell morphology

After each group of HK-2 cells are intervened for 48 hours, the result of biological inverted microscope analysis shows that the HK-2 cells in the normal group are in an obvious paving stone state, the cells in the model group are in obvious fibrosis, and the cells in the positive drug administration group and the traditional Chinese medicine composition capsule treatment group are close to the cells in the normal group.

(2) Influence on expression level of E-cadherin, α -SMA, p-ERK, NOX1, NOX2 and NOX4 proteins of HK-2 cells induced by high sugar.

TABLE 3 Effect of groups on the expression level of E-cadherin, α -SMA, FN, p-ERK, NOX1, NOX2, NOX4 proteins

Note: # was significantly different from the normal group. Significant difference P <0.05 compared to model control group;

the experimental results show that the traditional Chinese medicine composition has a remarkable hypoglycemic effect, and also has good effects of protecting kidney injury, improving renal tubular fibrosis and preventing and treating diabetic nephropathy. Can be used for treating diabetes or diabetic nephropathy.

Claims

1. The application of a traditional Chinese medicine composition in preparing a medicament for treating diabetes comprises radix astragali, fructus Lycii, rhizoma Dioscoreae, Coicis semen, radix scrophulariae, radix Glehniae, Ecliptae herba, placenta hominis, Eucommiae cortex, Notoginseng radix, herba Leonuri, Hirudo, periostracum Cicadae, semen plantaginis, and Achyranthis radix.

2. The traditional Chinese medicine composition comprises astragalus membranaceus, wolfberry fruits, Chinese yams, coix seeds, radix scrophulariae, radix glehniae, eclipta alba, human placenta, eucommia ulmoides, pseudo-ginseng, motherwort, leeches, cicada slough, semen plantaginis and radix achyranthis bidentatae.

3. The use according to claim 1 or 2, wherein the diabetes is type II diabetes.

4. The use of claim 3, wherein the extract of the Chinese medicinal composition is formulated with a pharmaceutically acceptable carrier in the form of a capsule, tablet, granule, pill or oral liquid.

5. The use as claimed in claim 3, wherein the extract of the Chinese medicinal composition is prepared by decocting the raw materials in water or extracting with ethanol under reflux, continuously refluxing, ultrasonically extracting, and concentrating.

6. The use of claim 1 or 2, wherein the Chinese medicinal composition is prepared from 680-1360 parts of radix astragali, 170-340 parts of fructus Lycii, 255-510 parts of rhizoma Dioscoreae, 170-340 parts of semen Coicis, 170-340 parts of radix scrophulariae, 170-340 parts of radix Glehniae, 170-340 parts of herba Ecliptae, 38-76 parts of placenta hominis, 340-680 parts of cortex Eucommiae, 153-306 parts of radix Notoginseng, 510-1020 parts of herba Leonuri, 38-76 parts of Hirudo, 51-102 parts of periostracum Cicadae, 170-340 parts of semen plantaginis, and 170-340 parts of radix Achyranthis bidentatae.

7. The use of claim 6, wherein the Chinese medicinal composition is prepared from 680 parts of astragalus mongholicus, 170 parts of wolfberry fruit, 255 parts of Chinese yam, 170 parts of coix seed, 170 parts of radix scrophulariae, 170 parts of radix glehniae, 170 parts of eclipta, 38 parts of human placenta, 340 parts of eucommia ulmoides, 153 parts of pseudo-ginseng, 510 parts of motherwort, 38 parts of leech, 51 parts of periostracum cicadae, 170 parts of semen plantaginis and 170 parts of radix achyranthis bidentatae.

8. The use of any one of claims 1 to 4, wherein the preparation method of the Chinese medicinal composition comprises:

(1) pulverizing Notoginseng radix, placenta hominis and Hirudo into fine powder, sieving, and sterilizing;

(2) decocting radix astragali, fructus Lycii, rhizoma Dioscoreae, Coicis semen, radix scrophulariae, radix Glehniae, Ecliptae herba, Eucommiae cortex, herba Leonuri, periostracum Cicadae, semen plantaginis, and Achyranthis radix in water, mixing decoctions, filtering, concentrating the filtrate under reduced pressure to obtain fluid extract, adding ethanol, stirring, standing, collecting supernatant, recovering ethanol under reduced pressure, and concentrating to obtain fluid extract; adding the fine powder of the pseudo-ginseng, the human placenta and the leech in the step (1), uniformly mixing, granulating, drying, crushing into fine powder, adding a pharmaceutically acceptable carrier, and preparing into a pharmaceutical preparation.

9. The use of claim 8, wherein the preparation method of the Chinese medicinal composition comprises:

(2) taking radix astragali, fructus lycii, Chinese yam, semen coicis, radix scrophulariae, radix glehniae, herba ecliptae, eucommia ulmoides, leonurus, periostracum cicadae, semen plantaginis and radix achyranthis bidentatae according to a weight ratio, adding water for decocting twice, 1 hour each time, combining decoction, filtering, concentrating filtrate under reduced pressure to obtain clear paste with the relative density of 1.16-1.20 at 60 ℃, adding ethanol to enable the alcohol content to reach 63%, stirring uniformly, standing for 24 hours, taking supernate, recovering ethanol under reduced pressure, and concentrating to obtain clear paste with the relative honey degree of 1.36-1.39 at 60 ℃; adding the fine powder of the pseudo-ginseng, the human placenta and the leech in the step (1), uniformly mixing, granulating, drying, crushing into fine powder, adding a pharmaceutically acceptable carrier, and preparing into a pharmaceutical preparation.