CN111643641B

CN111643641B - Food therapy formula for clearing heat, invigorating spleen and tonifying kidney and preparation method and application thereof

Info

Publication number: CN111643641B
Application number: CN202010651507.9A
Authority: CN
Inventors: 杨爱东; 朱烨琳; 朱华贺
Original assignee: Shanghai University of Traditional Chinese Medicine
Current assignee: Shanghai University of Traditional Chinese Medicine
Priority date: 2020-07-08
Filing date: 2020-07-08
Publication date: 2022-02-11
Anticipated expiration: 2040-07-08
Also published as: CN111643641A

Abstract

The invention discloses a food therapy formula for clearing heat, invigorating spleen and tonifying kidney and a preparation method and application thereof, wherein the food therapy formula for clearing heat, invigorating spleen and tonifying kidney is prepared by decocting raw materials of 1-10 parts by mass of sun-dried ginseng, 1-30 parts by mass of wrinkled gianthyssop herb, 1-30 parts by mass of rhizoma polygonati, 1-30 parts by mass of semen coicis, 1-60 parts by mass of Chinese yam, 1-30 parts by mass of wolfberry fruit, 1-30 parts by mass of bamboo leaf, 1-20 parts by mass of pepper, 1-30 parts by mass of lotus leaf, 1-30 parts by mass of cape jasmine fruit, 1-40 parts by mass of purslane, 1-30 parts by mass of fermented soybean, 1-20 parts by mass of galangal, 1-30 parts by mass of gorgon fruit, 1-20 parts by mass of dark plum, 1-20 parts by mass of turmeric, 1-40 parts by mass of radix puerariae, 1-30 parts by mass of radix polygonati officinalis, 1-30 parts by mass of mulberry leaf and 1-20 parts by mass of mint. Test results show that the food therapy formula can be used for treating diabetes, has a positive long-term curative effect and has an unexpected food therapy effect.

Description

Food therapy formula for clearing heat, invigorating spleen and tonifying kidney and preparation method and application thereof

Technical Field

The invention relates to a food therapy formula for clearing heat, invigorating spleen and tonifying kidney and a preparation method and application thereof, belonging to the technical field of food therapy.

Background

Diabetes is one of chronic diseases seriously harming human health, about 1.5 hundred million people suffer from diabetes in the world at present, while the number of diabetes in China accounts for about one third of the number of diabetes patients in the world, and the diabetes is the most popular country in the world, and the prevalence rate of the diabetes is increased year by year. Diabetes is a disease of endocrine disturbance in which glucose metabolism dysfunction is caused by multifactorial actions such as heredity and environment. Chronic hyperglycemia can cause a variety of diabetic complications, with diabetic nephropathy being the most common, and ultimately leading to renal failure, which is life threatening.

At present, researches show that the incidence rate of diabetic complications can be effectively reduced by controlling blood sugar, but various hypoglycemic drugs which are full of in the modern medical market are not only expensive, but also have certain dependence and tolerance of oral hypoglycemic drugs, and have obvious side effects such as diarrhea, inappetence, edema and the like after long-time administration. The traditional Chinese medicine has great advantages in the aspects of helping diabetes to improve clinical symptoms, delaying pathological progress, improving life quality and the like. In the traditional Chinese medicine, diabetes belongs to the category of diabetes, the pathogenesis of the diabetes lies in deficiency of spleen and kidney and accumulation of damp-heat, the treatment method mainly aims at tonifying spleen and kidney, clearing heat and promoting diuresis, the kidney is the congenital origin, the spleen and stomach are the acquired origin, and if the diabetes is insufficient, qi and blood are not sufficient, all diseases are born, the spleen and stomach are nourished, and healthy qi is sufficient, and the healthy qi is sufficient, so that the healthy qi supports healthy energy and eliminates pathogenic factors.

The food therapy is a precious cultural heritage of Chinese nationalities, the king of the medicine, the sun Simiao thinks that food has great significance for health preservation, disease treatment and disease prevention, the first food therapy writing of Qianjin prescription and food therapy of China is written, and according to the food therapy works of ' hot of cold people, cold of hot people ', nourishing of deficiency people and purging of excess people ', the mixed diet paying attention to syndrome differentiation thinking can effectively improve clinical symptoms and alleviate the side effects of western medicines, so that the health-care food is economical and practical. Traditional Chinese medicine diet therapy and modern medical comprehensive treatment based on the guidance of the traditional Chinese medicine syndrome differentiation theory are an important direction for treating diabetes, and medicinal materials required by the diet therapy are both medicinal and edible products, are easy to obtain and low in price, and are very suitable for popularization to the public. Therefore, it is very important to research the dietotherapy for treating diabetes.

Disclosure of Invention

Aiming at the problems and the requirements in the prior art, the invention aims to provide a food therapy formula for clearing heat, invigorating spleen and tonifying kidney and a preparation method and application thereof, and provides a food therapy approach for effectively treating diabetes.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

a food therapy formula for clearing heat, tonifying spleen and tonifying kidney is prepared by decocting 1-10 parts by mass of sun-dried ginseng, 1-30 parts by mass of wrinkled gianthyssop herb, 1-30 parts by mass of rhizoma polygonati, 1-30 parts by mass of coix seed, 1-60 parts by mass of Chinese yam, 1-30 parts by mass of wolfberry fruit, 1-30 parts by mass of bamboo leaf, 1-20 parts by mass of pepper, 1-30 parts by mass of lotus leaf, 1-30 parts by mass of gardenia, 1-40 parts by mass of purslane, 1-30 parts by mass of fermented soybean, 1-20 parts by mass of galangal, 1-30 parts by mass of gordon euryale seed, 1-20 parts by mass of dark plum, 1-20 parts by mass of turmeric, 1-40 parts by mass of kudzu root, 1-30 parts by mass of polygonatum, 1-30 parts by mass of mulberry leaf and 1-20 parts by mass of mint as raw materials.

Furthermore, the food therapy formula for clearing heat, tonifying spleen and tonifying kidney is prepared by decocting 2-5 parts by mass of sun-dried ginseng, 5-15 parts by mass of wrinkled gianthyssop herb, 5-15 parts by mass of rhizoma polygonati, 5-25 parts by mass of coix seed, 20-45 parts by mass of Chinese yam, 5-15 parts by mass of wolfberry fruit, 5-15 parts by mass of bamboo leaf, 5-10 parts by mass of pepper, 5-15 parts by mass of lotus leaf, 5-15 parts by mass of gardenia, 15-35 parts by mass of purslane, 5-15 parts by mass of fermented soybean, 5-10 parts by mass of galangal, 5-15 parts by mass of gordon euryale seed, 5-15 parts by mass of dark plum, 5-10 parts by mass of turmeric, 10-30 parts by mass of kudzu root, 5-15 parts by mass of polygonatum, 5-15 parts by mass of mulberry leaf and 5-10 parts by mass of mint as raw materials.

Furthermore, the food therapy formula for clearing heat, tonifying spleen and tonifying kidney is prepared by decocting 3 parts by mass of sun-dried ginseng, 12 parts by mass of wrinkled gianthyssop herb, 12 parts by mass of rhizoma polygonati, 15 parts by mass of semen coicis, 30 parts by mass of Chinese yam, 12 parts by mass of wolfberry fruit, 12 parts by mass of bamboo leaf, 6 parts by mass of pepper, 12 parts by mass of lotus leaf, 12 parts by mass of gardenia, 20 parts by mass of purslane, 12 parts by mass of fermented soybean, 6 parts by mass of galangal, 9 parts by mass of gordon euryale seed, 9 parts by mass of dark plum, 6 parts by mass of turmeric, 20 parts by mass of radix puerariae, 12 parts by mass of radix polygonati officinalis, 12 parts by mass of mulberry leaf and 6 parts by mass of mint as raw materials with water.

The medicinal materials in the food therapy formula for clearing heat, tonifying spleen and tonifying kidney are all homologous in medicine and food.

A preparation method of a food therapy formula for clearing heat, invigorating spleen and tonifying kidney comprises the steps of soaking all medicinal materials in water for 0.5-1.5 hours according to a ratio, decocting all the medicinal materials together with strong fire, boiling, and then slowly decocting for half an hour with slow fire; after removing the residue and taking the decoction, the method is continued, and the decoction is mixed twice.

Furthermore, in the preparation process, the water adding amount is controlled to be 2-3 cm higher than the medicine surface on the water surface.

An application of a dietetic therapy prescription for clearing heat, invigorating spleen and tonifying kidney, which is used for preparing a medicinal preparation for treating diabetes.

Further, the pharmaceutical preparation is an oral preparation.

Compared with the prior art, the invention has the following remarkable beneficial effects:

the test result shows that the food therapy formula has the effects of clearing heat, invigorating spleen and tonifying kidney, can effectively improve the symptoms of diabetic kidney and bladder injury, can be used for treating diabetes, has definite long-term curative effect, no side effect and lower cost, is suitable for long-term administration, and has popularization and application values.

Drawings

FIG. 1 is a graph showing the change in body mass of rats in example 5;

FIG. 2 is a graph showing the blood glucose changes in the tail vein of the rat in example 5;

FIG. 3 shows the effect of the formula of diet therapy for clearing heat, invigorating spleen and tonifying kidney on the pathological morphology of diabetic rat kidney tissue;

FIG. 4 shows the effect of the formula of diet therapy for clearing heat, invigorating spleen and tonifying kidney on pathological forms of bladder tissues of diabetic rats;

FIG. 5 shows the protein expression of TGF-beta/alpha-SMA and PD1/PDL in diabetic rat kidney tissues by the formula for clearing heat, invigorating spleen and tonifying kidney.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.

The raw materials of the sun-dried ginseng, the wrinkled gianthyssop herb, the sealwort, the coix seed, the Chinese yam, the medlar, the bamboo leaf, the pepper, the lotus leaf, the gardenia, the purslane, the fermented soybean, the galangal, the gordon euryale seed, the dark plum, the turmeric, the kudzuvine root, the fragrant solomonseal rhizome, the mulberry leaf, the mint and the like used in the invention all accord with the relevant provisions under the medicinal material items in the first part of the text of Chinese pharmacopoeia 2005 edition. The medicinal materials are cleaned, cut, processed, crushed and the like, and before the materials are put into the medicine, the material objects of the medicinal materials are identified, the names of the material objects are consistent, and the quality of the material objects accords with the standard of national pharmacopeia (the identification method of the specific medicinal materials is executed by referring to the standard of the pharmacopeia).

Example 1: preparation of diet therapy formula for clearing heat, invigorating spleen and tonifying kidney

Weighing 3g of sun-dried ginseng, 9g of wrinkled gianthyssop herb, 9g of rhizoma polygonati, 12g of semen coicis, 20g of Chinese yam, 10g of medlar, 12g of bamboo leaves, 5g of pepper, 10g of lotus leaves, 10g of gardenia, 20g of purslane, 12g of fermented soybean, 5g of galangal, 12g of gordon euryale seed, 6g of dark plum, 5g of turmeric, 15g of radix puerariae, 10g of radix polygonati officinalis, 12g of folium mori and 6g of mint; soaking all the medicinal materials in water for 0.5 hr, decocting all the medicinal materials with water over strong fire, boiling, and decocting with slow fire for half an hour; removing residues, collecting decoction, mixing the decoctions; the water adding amount is controlled to be 2-3 cm higher than the medicine surface in the preparation process.

Example 2: preparation of diet therapy formula for clearing heat, invigorating spleen and tonifying kidney

Weighing 3g of sun-dried ginseng, 12g of wrinkled gianthyssop herb, 9g of rhizoma polygonati, 15g of semen coicis, 30g of Chinese yam, 12g of medlar, 9g of bamboo leaves, 6g of pepper, 9g of lotus leaves, 12g of gardenia, 20g of purslane, 9g of fermented soybean, 6g of galangal, 9g of gordon euryale seed, 9g of dark plum, 6g of turmeric, 20g of radix puerariae, 12g of radix polygonati officinalis, 9g of folium mori and 6g of mint; soaking all the medicinal materials in water for 1 hr, decocting all the medicinal materials with water over strong fire, boiling, and decocting with slow fire for half an hour; removing residues, collecting decoction, mixing the decoctions; the water adding amount is controlled to be 2-3 cm higher than the medicine surface in the preparation process.

Example 3: preparation of diet therapy formula for clearing heat, invigorating spleen and tonifying kidney

Weighing 3g of sun-dried ginseng, 12g of wrinkled gianthyssop herb, 12g of rhizoma polygonati, 15g of semen coicis, 30g of Chinese yam, 12g of medlar, 12g of bamboo leaves, 6g of pepper, 12g of lotus leaves, 12g of gardenia, 20g of purslane, 12g of fermented soybean, 6g of galangal, 9g of gordon euryale seed, 9g of dark plum, 6g of turmeric, 20g of radix puerariae, 12g of radix polygonati officinalis, 12g of folium mori and 6g of mint; soaking all the medicinal materials in water for 1 hr, decocting all the medicinal materials with water over strong fire, boiling, and decocting with slow fire for half an hour; removing residues, collecting decoction, mixing the decoctions; the water adding amount is controlled to be 2-3 cm higher than the medicine surface in the preparation process.

Example 4: preparation of diet therapy formula for clearing heat, invigorating spleen and tonifying kidney

Weighing 3g of sun-dried ginseng, 12g of wrinkled gianthyssop herb, 9g of rhizoma polygonati, 12g of semen coicis, 25g of Chinese yam, 12g of medlar, 9g of bamboo leaves, 6g of pepper, 9g of lotus leaves, 12g of gardenia, 15g of purslane, 9g of fermented soybean, 6g of galangal, 12g of gordon euryale seed, 9g of dark plum, 6g of turmeric, 18g of radix puerariae, 9g of radix polygonati officinalis, 9g of folium mori and 5g of mint; soaking all the medicinal materials in water for 1.5 hr, decocting all the medicinal materials with water over strong fire, boiling, and decocting with slow fire for half an hour; removing residues, collecting decoction, mixing the decoctions; the water adding amount is controlled to be 2-3 cm higher than the medicine surface in the preparation process.

Example 5: pharmaceutical research of dietary therapy formula for clearing heat, invigorating spleen and tonifying kidney

5.1, animals:

healthy male Wistar clean grade rats 40, SPF grade, body mass (200 ± 20) g, shanghai slyke laboratory animals llc, certification No.: 20170005015466, special standard pellet feed, water feeding, animal ethics check number: PZSHUTCM190712005, experiments were performed in the experimental animal center facility of the university of medicine in shanghai (SYCK (shanghai) 2013-.

5.2, main equipment and instruments:

cryo-refrigerated centrifuge 3K15(Sigma, usa); microplate reader 680 (Bio-Rad, USA); SDS-PAGE electrophoresis apparatus (MINI Protean2 Bio-Rad, USA), Roche Excellent glucometer.

5.3, drugs and reagents:

metformin hydrochloride tablets, national pharmaceutical group chemical agents limited, lot No. 20181206; streptozotocin, sigma aldrich (shanghai) trade ltd, lot No.: WXBC 8740V; rabbit monoclonal PD1 antibody (ab243644), rabbit monoclonal PDL1 antibody (ab228462), rabbit monoclonal TGF-BETA antibody (ab179695) and rabbit monoclonal α SMA antibody (ab32575) were purchased from Abcam corporation (USA); anti-rabbit IgG (H + L) (8889S) was obtained from Cell Signaling, Inc. (USA); the ECL chemiluminescence solution and the BCA protein concentration determination kit are supported by Biyuntian research institute (Jiangsu province); EnVision reagent (HRP/rabbit) was purchased from Dako corporation (denmark);

the food therapy formula for clearing heat, tonifying spleen and tonifying kidney prepared in example 3 is prepared by heating and concentrating decoction liquid to 123.5mL, namely, the concentration is 2g/mL, cooling and storing at 4 ℃ for later use.

5.4, a statistical method:

the experimental results are all adopted

Showing that SPSS 21.0 statistical software is adopted for all data, single-factor analysis of variance is adopted for comparison among groups, and P is less than 0.05, which is of statistical significance.

5.5, general state observation of rats:

the experimental method comprises the following steps:

selecting 40 healthy male Wistar cleaning-grade rats, and randomly dividing the rats into a normal group, a model group, a diet therapy formula (high and low dose) group for clearing heat, tonifying spleen and tonifying kidney and a metformin group, wherein each group comprises 8 rats; the weight is 200 +/-20 g, and the normal group is fed with the common feed after the healthy male Wistar clean-grade rats are adaptively fed for one week; the rest rats are forbidden to take the night, 0.1mol/L citric acid buffer solution (pH4.0) is used for preparing 1% concentration before Streptozotocin (STZ) is used, 55mg/kg dosage is used for intraperitoneal one-time injection, fasting blood sugar is detected after 72 hours, only equivalent citric acid buffer solution is injected in an abdominal cavity in a normal group, and blood sugar detected for three times continuously is used as a model for selection; the successfully molded diabetes rat models are randomly divided into a model group, a metformin group and a high and low dose group of a food therapy prescription for clearing, tonifying spleen and tonifying kidney; after the molding is successful, the metformin and a low-dose and high-dose food therapy prescription for clearing heat, tonifying spleen and tonifying kidney are respectively administrated for intragastric administration for 1 time/day (the metformin is 100mg/kg, the low-dose food therapy prescription is 4.5g/kg, and the high-dose food therapy prescription is 18g/kg) for 4 weeks; observing all rats in the experimental period, wherein the mental state, the activity condition, the fur glossiness, the death condition and the like are observed, measuring the weight and the blood sugar of the rats every 7 days, after four weeks, fasting the rats for 6 hours, weighing, after anesthesia, killing each group of rats, and opening the chest and abdomen to collect kidney tissues and bladder tissues; part of the cells were fixed with 4% paraformaldehyde and stored at-80 ℃ and the histopathological morphological changes of kidney and bladder were observed under a light microscope, while the rest were analyzed by Western blot.

The experimental results are as follows:

during the experiment 2 rats in the model group died; the performance before and after administration is as follows: after the streptozotocin is administrated by gastric lavage for a period of time, the rats in the normal group show that the body form is obviously increased, the tail vein blood sugar is normal, the fur is dry and smooth, the mental state is good, and the rats are active and active compared with the rats in the model group and the administration groups; the blood sugar of the rats in the model group is continuously higher and gradually shows symptoms of spleen and kidney deficiency, such as the rats are gradually thinned, hair is fallen, hair loses luster, listlessness, slow response, the amount of drinking water is reduced, and activities are obviously reduced; the rats in the normal control group have no abnormal phenomena in the aspects of body mass, blood sugar, food intake, various reactions and the like; during the treatment administration period of each administration group, after the administration by gastric lavage, the symptoms of the deficiency of the spleen and the kidney are all relieved to a certain degree, and the blood sugar is controlled to a certain degree.

Specifically, the change in body mass of each group of rats is shown in table 1 and fig. 1.

TABLE 1 change in body Mass (g) of rats in each group: (

n＝6)

In table 1, # compared to the normal group: p <0.05, # #: p < 0.01; comparison with model group,: p <0.05, x: p < 0.01; compared with the metformin group, the metformin hydrochloride is in good coverage: p <0.05, in the color: p < 0.01.

FIG. 1 is a graph showing the change in body mass of rats in this example. As can be seen from Table 1 and FIG. 1, the body mass of the model group rats was significantly reduced on days 1, 14 and 28 of treatment (P <0.01) compared with the normal group, and there was no significant difference in body mass between the treatment groups on days 1 and 14 of treatment compared with the model group; on day 28 of treatment, the body mass increased significantly for each treatment group (P < 0.05).

The change in body mass of each group of rats is shown in table 2 and fig. 2.

TABLE 2 Tail vein blood glucose Change (mmol/L) of rats in groups: (

n＝6)

Group of	Day 1	Day 7	Day 14	Day 21	Day 28
						Normal group	5.4±0.16	5.5±0.16	5.6±0.15	5.5±0.18	5.6±0.24
Model set	20.9±1.00^##	20.8±0.81	21.3±0.89^##	20.9±1.29	21.5±1.83^##
						Metformin hydrochloride	21.4±1.22	18.4±0.76	17.3±0.92^*	14.1±0.81	12.2±1.94^**
Low dose group	21.4±1.13	19.9±0.98	19.1±1.02	17.1±1.71	16.8±2.27^*※
						High dose group	20.5±0.52	18.2±0.68	16.9±0.92^*	13.1±1.44	11.9±1.98^**

In table 2, # compared to the normal group: p <0.05, # #: p < 0.01: (ii) a Comparison with model group,: p <0.05, x: p < 0.01; comparison with metformin group: in addition: p <0.05, in the color: p < 0.01.

FIG. 2 is a graph showing the blood glucose changes in the tail vein of the rat in this example. As can be seen from table 2 and fig. 2, the tail vein blood glucose levels of the rats in the model group were significantly increased on days 1, 14 and 28 of treatment (P <0.01) compared with the normal group, and the tail vein blood glucose levels of the rats in the high and low dose groups were not significantly different between the metformin group and the qingli-tonifying and kidney-tonifying diet therapy on day 1 of treatment, the tail vein blood glucose levels of the rats in the high dose group and the qingli-tonifying and kidney-tonifying diet therapy on day 14 of treatment (P <0.05), and the tail vein blood glucose levels of the rats in the high and low dose groups of the metformin group and the qingli-tonifying and kidney-tonifying diet therapy on day 28 of treatment (P <0.01, P <0.05) compared with the normal group.

5.6 pathological morphology observation of rat kidney and bladder

The experimental method comprises the following steps:

rat kidney and bladder tissues were taken and fixed in 4% paraformaldehyde and stored at-80 ℃. Dehydrating the paraffin sections conventionally, and washing with PBS for 3X 3 minutes; thermally induced with citric acid buffer (CB) pH 6.0 (microwave 3 for 20 min, natural cooling at room temperature, washing with PBS for 3X 3 min; 0.3% H)₂O₂Inhibiting endogenous peroxidase for 20 minutes at room temperature; washing with PBS for 3 × 3 min; incubating 20% normal sheep serum for 30 min at room temperature; adding primary antibody dropwise and incubating for 2 hours at 37 ℃; PBS wash for 3X 3 min; EnVision reagent (HRP/R)37 ℃, 30 minutes; PBS wash for 3X 3 min; DAB color development is carried out for 8-12 minutes; hematoxylin-eosin staining, dehydrating, clearing, sealing, and observing pathological changes of kidney and bladder tissues under an optical microscope.

Results of the experiment

1) HE staining observation of rat kidney tissue

FIG. 3 shows the effect of the formula of diet therapy for clearing heat, invigorating spleen and tonifying kidney on the pathological morphology of diabetic rat kidney tissue; as can be seen from FIG. 3, no significant pathological changes were observed in normal renal tissues; the epithelial cells of the far-end renal tubule of the model group part are obviously swelled and necrotic, glomerular crescent is formed, the local far-end near-end renal tubule is expanded, and the epithelial cells of the wall layer of the glomerular capsule are locally proliferated; partial renal tubular watery degeneration of the metformin group, local distal and proximal renal tubular dilatation, epithelial cell swelling and necrosis; a small amount of distal tubular epithelial cells are obviously swollen (i.e. balloon-like) and proximal tubular epithelial cells are expanded in a low-dose group part of the diet therapy formula for clearing heat, tonifying spleen and tonifying kidney; part of local distal renal tubular epithelial cells of a high-dose group of the diet therapy formula for clearing heat, tonifying spleen and tonifying kidney obviously swell, and proximal renal tubular cells expand.

2) HE staining observation of rat bladder

FIG. 4 shows the effect of the formula of diet therapy for clearing heat, invigorating spleen and tonifying kidney on pathological forms of bladder tissues of diabetic rats; as can be seen in FIG. 4, no significant pathological changes were observed in the normal group of bladder wall tissues; the model group rat bladder wall smooth muscle cell part is hypertrophied, the form is various, the arrangement is disordered, the muscle cell is slightly mucus-like degeneration; partial hypertrophy, various forms, disorganization and local intercellular mucus-like degeneration of the smooth muscle cells of the bladder wall in the metformin group; the part of the low-dose group of the diet therapy prescription for clearing heat, tonifying spleen and tonifying kidney can be seen to be slightly hypertrophied by smooth muscle cells of bladder wall, and the forms are various; the hypertrophy arrangement disorder of the muscle cells of the bladder wall can be seen in the high-dose group of the food therapy formula for clearing heat, tonifying spleen and tonifying kidney.

5.7 determination of TGF-. beta./alpha. -SMA/PD1/PDL1 expression in rat Kidney tissue

Experimental methods

Kidney tissue was taken and homogenized to extract proteins, and then equal amounts of the obtained proteins were subjected to 6% SDS-PAGE electrophoresis by the bicinchoninic acid (BCA) method, then constant voltage transmembrane for 180 minutes at 100V then primary antibody (for PD1, PDL1, TGF- β or α -SMA) and incubated overnight at 4 ℃, membrane washed with TBST, then diluted secondary antibody added for 1 hour again with TBST, then gel imaging analysis system was used for analysis, grey values were analyzed using AlphaView SA software (protein simple, CA, USA), i.e. for quantitative analysis, grey scale calculation from grey scale target bar to inner bar was referenced as relative expression level target protein.

Results of the experiment

FIG. 5 shows the protein expression of TGF-beta/alpha-SMA and PD1/PDL in diabetic rat kidney tissues by the dietotherapy for clearing heat, invigorating spleen and tonifying kidney; in the figure, # compared to the normal set: p <0.05, # #: p < 0.01; comparison with model group,: p <0.05, x: p < 0.01; comparison with metformin group: in addition: p <0.05, in the color: p < 0.01; as can be seen in FIG. 5, TGF- β and α -SMA expression was significantly increased in rat kidney tissue in the model group compared to the normal group (P < 0.01). Compared with the model group, the expression of TGF-beta and alpha-SMA in the kidney tissues of rats in high and low dose groups of the food therapy formula for clearing heat, tonifying spleen and tonifying kidney is obviously reduced (P is less than 0.01).

In conclusion, the food therapy formula can obviously improve the states of the diabetic rat, such as mental state, activity condition, fur glossiness and death condition, can obviously improve the pathological form of the kidney tissue of the diabetic rat, and can reduce the damage of the bladder tissue to be not obviously different from that of a normal group; the expression of TGF-beta and alpha-SMA in kidney tissues of rats with renal diabetes can be obviously inhibited, and the immune response of an organism can be regulated, so that the dietary therapy formula provided by the invention can be used for treating diabetes, and has obvious progress and unexpected effect compared with the prior art.

Finally, it should be pointed out here that: the above is only a part of the preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention, and the insubstantial modifications and adaptations of the present invention by those skilled in the art based on the above description are intended to be covered by the present invention.

Claims

1. A food therapy formula for clearing heat, invigorating spleen and tonifying kidney is characterized in that: the health care tea is prepared by decocting 1-10 parts by mass of sun-dried ginseng, 1-30 parts by mass of wrinkled gianthyssop herb, 1-30 parts by mass of rhizoma polygonati, 1-30 parts by mass of semen coicis, 1-60 parts by mass of Chinese yam, 1-30 parts by mass of wolfberry fruit, 1-30 parts by mass of bamboo leaf, 1-20 parts by mass of pepper, 1-30 parts by mass of lotus leaf, 1-30 parts by mass of cape jasmine fruit, 1-40 parts by mass of purslane, 1-30 parts by mass of fermented soybean, 1-20 parts by mass of galangal, 1-30 parts by mass of gordon euryale seed, 1-20 parts by mass of dark plum, 1-20 parts by mass of turmeric, 1-40 parts by mass of radix puerariae, 1-30 parts by mass of radix polygonati officinalis, 1-30 parts by mass of mulberry leaf and 1-20 parts by mass of mint as raw materials.

2. The dietary therapy formula for clearing heat, invigorating spleen and tonifying kidney according to claim 1, characterized in that: the health care tea is prepared by decocting raw materials of 2-5 parts by mass of sun-dried ginseng, 5-15 parts by mass of wrinkled gianthyssop herb, 5-15 parts by mass of rhizoma polygonati, 5-25 parts by mass of semen coicis, 20-45 parts by mass of Chinese yam, 5-15 parts by mass of wolfberry fruit, 5-15 parts by mass of bamboo leaf, 5-10 parts by mass of pepper, 5-15 parts by mass of lotus leaf, 5-15 parts by mass of cape jasmine fruit, 15-35 parts by mass of purslane, 5-15 parts by mass of fermented soybean, 5-10 parts by mass of galangal, 5-15 parts by mass of gordon euryale seed, 5-15 parts by mass of dark plum, 5-10 parts by mass of turmeric, 10-30 parts by mass of radix puerariae, 5-15 parts by mass of polygonatum, 5-15 parts by mass of mulberry leaf and 5-10 parts by mass of mint.

3. The dietary therapy formula for clearing heat, invigorating spleen and tonifying kidney according to claim 1 or 2, characterized in that: is prepared by decocting 3 parts by mass of sun-dried ginseng, 12 parts by mass of wrinkled gianthyssop herb, 12 parts by mass of rhizoma polygonati, 15 parts by mass of semen coicis, 30 parts by mass of Chinese yam, 12 parts by mass of medlar, 12 parts by mass of bamboo leaves, 6 parts by mass of pepper, 12 parts by mass of lotus leaves, 12 parts by mass of gardenia, 20 parts by mass of purslane, 12 parts by mass of fermented soybean, 6 parts by mass of galangal, 9 parts by mass of gordon euryale seed, 9 parts by mass of dark plum, 6 parts by mass of turmeric, 20 parts by mass of radix puerariae, 12 parts by mass of polygonatum odoratum, 12 parts by mass of mulberry leaves and 6 parts by mass of mint as raw materials.

4. A preparation method of the dietary therapy prescription for clearing heat, invigorating spleen and tonifying kidney according to claim 1, which is characterized in that: soaking all the medicinal materials in water for 0.5-1.5 hours according to the proportion, decocting all the medicinal materials together with water over strong fire until boiling, and then decocting slowly over slow fire for half an hour; after removing the residue and taking the decoction, the method is continued, and the decoction is mixed twice.

5. The method of claim 4, wherein: in the preparation process, the water adding amount is controlled to be 2-3 cm higher than the medicine surface on the water surface.

6. The application of the food therapy formula for clearing heat, invigorating spleen and tonifying kidney according to claim 1 is characterized in that: is used for preparing a medicinal preparation for treating diabetes.

7. Use according to claim 6, characterized in that: the pharmaceutical preparation is an oral preparation.