Application of laetiporus cinnabarina ttys 997 in preparation of medicine for treating or adjunctively treating diabetic nephropathy

Abstract

The invention provides application of laetiporus cinnabarina ttys 997 in preparation of a medicine for treatment or adjuvant treatment of diabetic nephropathy. The research finds that the laetiporus cinnabarina ttys 997 can be used for reducing the kidney index, the 24-hour urination volume and the urine microalbumin excretion rate of a rat with diabetic nephropathy; meanwhile, the content of urea nitrogen (BUN) and creatinine (Cr) in serum can be reduced, and the symptoms of foamy urine, nocturia increase, fatigue and the like of patients with diabetic nephropathy can be improved.

Description

Technical Field

[0001] The invention relates to application of sulphur fungus ttyS-997 in preparing medicine for treating or assisting in treating diabetic nephropathy. Background Art

[0002] Diabetic nephropathy (DN) is the main microvascular complication of diabetes. The prevalence of diabetic nephropathy in patients with type 2 diabetes in my country has reached 40%. In the early pathological stage, the creatinine clearance capacity decreases, the serum urea nitrogen (BUN) and creatinine (Cr) levels increase, proteinuria occurs, kidney cells proliferate and hypertrophy, the extracellular matrix increases, and in the late stage, glomerular sclerosis and tubular interstitial fibrosis occur, and eventually renal failure occurs. Diabetic nephropathy is currently the main cause of end-stage renal failure worldwide. The Western medicine treatment of diabetic nephropathy mainly involves adjusting lifestyle, lowering blood sugar, lowering blood pressure, regulating lipids, and reducing proteinuria. In the later stage, when kidney function gradually deteriorates, blood purification and kidney transplantation can be performed, and the methods are relatively limited. Therefore, there is an urgent need to develop new and effective kidney protection methods to alleviate diabetic nephropathy.

[0003] A large number of clinical studies have confirmed that traditional Chinese medicine has certain efficacy and advantages in treating diabetic nephropathy. Sulfur fungus, also known as forest chicken, is a rare edible and medicinal fungus that grows on logs, stumps and trunks. It is thick and delicious when young. The book "Compendium of Materia Medica" calls this fungus "Huangzhi". It can treat "five evils of the heart and stomach", temper and calm the mind, making people honest, kind and approachable. Regular consumption can make people light and not old, and prolong their life to become immortal. The Tujia people have used it to treat colitis, rheumatism and rheumatoid arthritis for thousands of years. It is widely used by the people abroad for fever, cough, gastric cancer and rheumatism. Due to the difficulty in collecting wild mushrooms and the small yield, it is expensive and cannot be widely used. Vermilion Sulfur Bacteria ttys-997 is a patented strain produced by our company using industrial culture technology (patent authorization number: ZL 202110330475.7 A Vermilion Sulfur Bacteria and its application in the treatment of gastrointestinal ulcers and colitis, ZL 202110288535.3 A Vermilion Sulfur Bacteria and its application). We have previously found that it can reduce blood sugar concentration, reduce the area of ​​diabetic foot wounds, significantly reduce the concentration of inflammatory factors TNF-α and IL-6 in the damaged skin tissue, increase the expression of vascular endothelial growth factor (VEGF) in the damaged skin tissue, accelerate the wound healing of diabetic rats, and has the effect of treating diabetic foot (patent application number: 202211059601.0 The application of Vermilion Sulfur Bacteria in the preparation of drugs, foods or health products for the treatment of diabetic foot). There is currently no research report that it can improve diabetic nephropathy. Summary of the invention

[0004] The purpose of the present invention is to provide the use of Sulphurum vermilion ttyS-997 in preparing medicine for treating or assisting in treating diabetic nephropathy.

[0005] To achieve the above objectives, the technical solutions adopted are:

[0006] The present invention provides the use of Laetiporus sulphureus ttys-997 in the preparation of a drug for the treatment or auxiliary treatment of diabetic nephropathy. The Laetiporus sulphureus (Laetiporus sulphureus) ttys-997 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on December 3, 2020. The deposit address is: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 21084. The Laetiporus sulphureus ttys-997 described in the present invention is an existing known strain, and the strain itself has been protected in a patent application number of 202110288535.3.

[0007] The invention provides application of culture supernatant of Sulphurum vermilionum ttyS-997 in preparing medicine for treating or assisting in treating diabetic nephropathy.

[0008] The invention provides application of a lysate of Sulphurum vermilionum ttyS-997 in preparing a medicine for treating or assisting in treating diabetic nephropathy.

[0009] The invention provides application of metabolites of Sulphurum vermilionum ttyS-997 in preparing medicine for treating or assisting in treating diabetic nephropathy.

[0010] Preferably, the drug comprises Sulphurum vermilion ttyS-997 in a pharmaceutically acceptable carrier.

[0011] Preferably, the drug is in the form of tablets, capsules, oral preparations or lyophilized powders.

[0012] Preferably, the pharmaceutically acceptable carrier includes at least one of skim milk, lactose, glucose, sucrose, sorbitol, mannose, trehalose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0013] The invention provides use of vermilion sulphur fungus ttys-997, culture supernatant of vermilion sulphur fungus ttys-997, lysate of vermilion sulphur fungus ttys-997 or metabolites of vermilion sulphur fungus ttys-997 in preparing food or health products.

[0014] Beneficial effects:

[0015] The present invention has found through research that the thylacinus ttyS-997 can reduce the renal index, 24-hour urine output, and urine microalbumin excretion rate of diabetic nephropathy rats; at the same time, it can reduce the content of urea nitrogen (BUN) and creatinine (Cr) in serum, and improve the symptoms of foamy urine, increased nocturia, fatigue, etc. in patients with diabetic nephropathy. DETAILED DESCRIPTION

[0016] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0017] Example 1

[0018] 1. Animal experiment: Experiment on improving diabetic nephropathy in rats with thymus vermilion sulphureus ttyS-997

[0019] 1. Methods

[0020] 1.1 Animal model construction and grouping

[0021] Healthy SD rats (male, body weight 200±20)g, SPF grade, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., certificate number SCXK (Beijing) 2007-0001. ), fasting but not water for 12h, once intraperitoneal injection of streptozotocin (Sigma, USA) 60mg / kg (based on body weight). After 72h, blood was collected from the tail vein and fasting blood glucose was measured. The model was successfully established when the blood glucose value was ≥16.7mmol / L. The normal control group was injected with an equal volume of citrate buffer (pH4.3). The rats with successful modeling were randomly divided into a model control group and a vermilion sulphur group according to body weight and blood glucose, with 10 rats in each group. The vermilion sulphur group was given 20ml / kg liquid preparation by gavage, and the normal control group and model control group were given normal saline with a gavage volume of 1mL / 100g for a total of 8 weeks.

[0022] Preparation of the liquid preparation of Vermilion Sulfur Bacteria ttyS-997:

[0023] The culture medium of Vermilion Sulfur Bacteria ttys-997: potato boiled water 15-30% (weight percentage), peptone 2-4% (weight percentage), glucose 10-25% (weight percentage), soybean powder 10-15% (weight percentage), beef extract 1-3% (weight percentage), potassium dihydrogen sulfate 0.1-0.3% (weight percentage), distilled water 1L, pH natural.

[0024] (1) Adding agar to the above liquid culture medium to prepare a slant culture medium, inoculating Vermilion Sulfur Bacillus ttyS-997, and culturing at 25-28° C. for 5-8 days to obtain a culture; inoculating the fresh culture into the above liquid culture medium, culturing at 22-25° C. for 4-5 days to obtain a seed solution; and then inoculating the seed solution into a fermentation tank at a concentration of 5-10% (weight percentage).

[0025] (2) Put pure water into a fermentation tank, add the above culture medium components, heat at high temperature and maintain at 100-120° C. for 20-30 minutes, cool to room temperature, inoculate at 5-10% (weight percentage), and culture at 20-32° C. with stirring for 4-6 days until the bacterial concentration reaches 30-40% (weight percentage) and the pH is 2-3. The tank is placed at room temperature to obtain a fermentation liquid.

[0026] (3) Place the fermentation liquid in a centrifuge at 3000-5000 rpm and centrifuge for 30 minutes to obtain a supernatant. Mix the supernatant with purified water and dilute it to a pH of 5-6. Place it in a sterilizing pot and heat it at a high temperature of 113-115°C for 30 minutes to obtain a liquid preparation after sterilization.

[0027] 1.2 Detection indicators

[0028] 1.2.1 After the last administration, the rats were subjected to 24-hour urine collection, centrifugation and packaging, and stored at 4°C for testing. After the last administration, the rats were fasted but not watered for 5 hours, and 100 μL of blood was collected from the tail vein. The supernatant was separated according to the kit method, and the blood glucose level was measured. After the kidneys were irrigated with 4°C precooled saline through the heart cannula until the kidneys turned white, the kidneys were removed and the capsules were removed and weighed. 8 mm3 of renal cortex was taken on ice and fixed in neutral formaldehyde for immunohistochemical staining. The remaining renal cortex was cut into small pieces and put into liquid nitrogen for tissue RNA and protein extraction.

[0029] 1.2.2 Determination of microalbumin content in urine

[0030] The urine microalbumin content was detected according to the kit (Shanghai Sun Biotechnology Co., Ltd.), and the urine microalbumin excretion rate (UAER) was the product of urine microalbumin and 24-hour urine volume.

[0031] 1.2.3 Determination of serum urea nitrogen (BUN) and creatinine (Cr) content

[0032] The levels of BUN, Cr and UA in rat serum were detected according to the kit method (Nanjing Jiancheng Bioengineering Institute).

[0033] 1.3 Statistical methods

[0034] The data were analyzed using SPSS 21.0 analysis software, and the experimental results were expressed as mean ± standard deviation (mean ± SD). One-way ANOVA was used to evaluate the overall variance difference between the means of multiple groups, and LSD was used for multiple comparisons. For comparisons between two groups, unpaired t-tests were used for comparison. P < 0.05 was considered to be significantly different.

[0035] 2. Results

[0036] Table 1 Effect of the culture medium of Sulphurum vermilionum ttyS-997 on diabetic nephropathy

[0037]

[0038] #. Compared with the normal control group, the difference is extremely significant (P<0.01); *. Compared with the model control group, the difference is significant (P<0.05); **. Compared with the model control group, the difference is extremely significant (P<0.01)

[0039] As shown in Table 1, the renal index of rats in the model control group was higher than that in the normal control group (P < 0.01). After oral administration of ttys-997, the renal index decreased by 27.67% (P < 0.01). This shows that ttys-997 has a good function of inhibiting renal hypertrophy. The 24-hour urine volume and UAER excretion rate of the model control group were significantly higher than those of the normal control group (P < 0.01). After oral administration of ttys-997, the 24-hour urine volume and UAER excretion rate of rats were reduced by 37.53% and 43.829% respectively compared with the model control group (P < 0.01). The blood BUN and Cr in the model control group were significantly higher than those in the normal control group (P < 0.01). After oral administration of ttys-997, the BUN and Cr of rats were reduced by 20.48% and 22.5% respectively compared with the model control group (P < 0.05). The above experimental results indicate that ttys-997 has a certain protective effect on diabetic nephropathy.

[0040] 2. Clinical observation: Sulphur fungus ttyS-997 improves renal function in patients with diabetic nephropathy

[0041] Four patients diagnosed with type 2 diabetes and diabetic nephropathy took 50 ml / day of thymol ttyS-997 oral solution for one month.

[0042] Table 2 Effects of Sulphurum vermilionum ttyS-997 on patients with diabetic nephropathy

[0043]

[0044] After the intervention, the patient's serum urea nitrogen (BUN), creatinine (Cr) and urine microalbumin decreased, and the subjective symptoms of diabetic nephropathy such as foamy urine, increased nocturia and fatigue were alleviated.

[0045] Conclusion: The culture solution of the thysinus vermilionii ttys-997 provided by the present invention can significantly reduce the renal index, 24h urine output, and urine microalbumin excretion rate of diabetic rats; at the same time, it can reduce the content of urea nitrogen (BUN) and creatinine (Cr) in serum, and improve the symptoms of foamy urine, increased nocturia, fatigue, etc. in patients with diabetic nephropathy. It has a certain improvement effect on diabetic nephropathy.

[0046] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. Use of Sulphurum vermilionum ttyS-997 in the preparation of drugs for treating or assisting in the treatment of diabetic nephropathy.

2. Application of the culture supernatant of Sulphur Fungus ttyS-997 in the preparation of drugs for the treatment or auxiliary treatment of diabetic nephropathy.

3. Use of the lysate of Sulphurum vermilionum ttyS-997 in the preparation of drugs for the treatment or auxiliary treatment of diabetic nephropathy.

4. Application of the metabolites of Sulphur Fungus ttyS-997 in the preparation of drugs for the treatment or auxiliary treatment of diabetic nephropathy.

5. The use according to claim 1, characterized in that: The drug comprises thylacinus tys-997 in a pharmaceutically acceptable carrier.

6. The use according to claim 1, characterized in that: The dosage form of the drug is tablet, capsule, oral preparation or lyophilized powder.

7. The use according to claim 2, characterized in that: The pharmaceutically acceptable carrier includes at least one of skim milk, lactose, glucose, sucrose, sorbitol, mannose, trehalose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.

8. Use of Sulphur Fungus ttyS-997, culture supernatant of Sulphur Fungus ttyS-997, lysate of Sulphur Fungus ttyS-997 or metabolites of Sulphur Fungus ttyS-997 in the preparation of food or health products.

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