Application of fungus strain Cs-2025 in preparation of medicine for treating renal function impairment

Abstract

The invention discloses an application of a fungus strain Cs-2025 in preparation of a medicine for treating renal function impairment, the fungus strain Cs-2025 is a new strain which is obtained from a natural cordyceps sinensis sporocarp through isolated culture and can be industrially produced, and the strain Cs-2025 is preserved in China General Microbiological Culture Collection Center, the preservation number is CGMCC (China General Microbiological Culture Collection Center) The medicine for treating renal function impairment provided by the invention comprises a culture of the fungus strain Cs-2025, and experimental research results show that the Cs-2025 has a good treatment effect on mouse renal function impairment caused by adenine, has the advantages of low toxicity, safety, effectiveness and the like, and can be used for preparing a novel medicine for resisting renal injury.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbial medicine, in particular to a new use of a fungal strain Cs-2025 in kidney injury protection. BACKGROUND

[0002] Cordyceps sincesis (Berk.) Sace. of Cordyceps family, and the compound of the larva corpse of the host insect Hepialus armoricanus Oberthur of Hepialidae, has a protective effect on kidney injury caused by various reasons, including drug-induced nephrotoxicity and glomerular sclerosis caused by various kidney diseases, can significantly improve kidney function, promote the repair and regeneration of renal tubules, and correct metabolic disorders. Studies have shown that some strains isolated from Cordyceps sincesis also have similar effects to Cordyceps sincesis.

[0003] Ophiocordyceps qingensis (Cs-2025) is a new strain isolated from natural Cordyceps sincesis in Aba region of Qinghai-Tibet Plateau in China, which has been preserved by China General Microbiological Culture Collection Center (Preservation Number: CGMCC NO. 41879, Preservation Date: April 2, 2025). There is no research report on the use of Cs-2025 in the treatment of kidney function injury. SUMMARY

[0004] The purpose of the present application is to provide the use of a fungal strain Cs-2025 in the preparation of a drug for treating kidney function injury.

[0005] In order to achieve the above-mentioned purpose, according to the first aspect of the present application, the use of a fungal strain Cs-2025 in the preparation of a drug for treating kidney function injury is provided, and the strain Cs-2025 is preserved in China General Microbiological Culture Collection Center, with a preservation number of CGMCC NO. 41879.

[0006] According to another aspect of the present application, a drug for treating kidney function injury is provided, which comprises the culture of the fungal strain Cs-2025 of claim 1.

[0007] Further, the drug treats kidney function injury by reducing purine levels.

[0008] According to another aspect of the present application, a preparation method of the culture of the fungal strain Cs-2025 is provided, which inoculates the mother strain of the strain Cs-2025 into a culture medium for culture, and then dries the culture medium after the culture is completed to obtain the culture.

[0009] Further, the culture medium comprises millet 180-220 grams, sucrose 8-12 grams, magnesium sulfate 200-300 milligrams, potassium dihydrogen phosphate 400-600 milligrams, and distilled water 400-600 milliliters in terms of mass fraction.

[0010] Further, the culture medium comprises millet 200 grams, sucrose 10 grams, magnesium sulfate 250 milligrams, potassium dihydrogen phosphate 500 milligrams, and distilled water 500 milliliters in terms of mass fraction.

[0011] Further, the pH value of the culture medium is 6.0-7.0.

[0012] Further, the temperature during the culture is controlled at 22-26°C.

[0013] Further, the relative humidity during the culture is 30-80%.

[0014] Further, the freeze-drying or drying under the temperature condition of 60-80°C.

[0015] The fungal strain Cs-2025 described in the present application is a new strain that can be industrialized for production and is obtained by isolation and culture from natural Cordyceps sinensis fruiting bodies. The experimental research results show that Cs-2025 has a good therapeutic effect on the renal function damage of mice caused by adenine, has small toxicity, is safe and effective, and can be used for preparing a new anti-renal injury drug. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 , Figure 2 and Figure 3 are contrasted kidney pathological pictures (HE staining ×200) related to Example 4, wherein, Figure 1 is a normal control group kidney pathological picture, Figure 2 is a model group, Figure 3 is a Cs-2025 dose group. Figure 2 In the pictures, A shows that the renal tubules are degenerated and necrotic, and inflammatory cell infiltration foci appear, B shows that the renal tubules are severely dilated, and the renal tubular epithelium is edematous. DETAILED DESCRIPTION

[0017] In order to more clearly understand the above-mentioned purposes, features and advantages of the present application, the schemes of the present application will be further described below. It should be noted that the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.

[0018] In the following description, a lot of specific details are set forth in order to give a thorough understanding of the present application, but the present application can also be implemented in other manners different from those described herein; obviously, the embodiments in the description are only some embodiments of the present application, not all the embodiments.

[0019] The preferred embodiments of the present application will be described in detail below with reference to Examples. It should be understood that the following Examples are given to generally illustrate the application and should not be considered as limiting the scope of the application. Various modifications and alterations of the present application can be made by those skilled in the art without departing from the spirit and scope of the application.

[0020] The experimental methods used in the following examples are conventional methods unless otherwise specified.

[0021] The materials, reagents, etc. used in the following examples can be obtained commercially unless otherwise specified.

[0022] The strain Cs-2025 involved in the present embodiment is Pseudomonas asiatica, which was deposited with the China General Microbiological Culture Collection Center (address: No. 1, Huayuancun, Beijing, China) on April 2, 2025, and has the accession number CGMCC NO. 41879. The strain has been disclosed in the Chinese patent application CN120591112A entitled “Strain for enhancing immune function and application thereof”.

[0023] Example 1 Preparation of Cs-2025 culture The formula of the culture medium is: millet 200 g, sucrose 10 g, magnesium sulfate 250 mg, potassium dihydrogen phosphate 500 mg, and distilled water 500 mL.

[0024] Preparation of the culture medium: Dissolve all ingredients except millet in water, adjust the pH value to 6.5, add rice, continue to heat and stir constantly, stop heating when it becomes solid, and then pour it into 10 wide-mouth bottles with a capacity of 500 mL, respectively, and seal with a layer of parchment paper. Sterilize in a high-pressure steam sterilization pot at 121°C for 20 minutes, cool to below 27°C, and then use.

[0025] Inoculation and culture: Open the culture bottle under sterile conditions, inoculate the mother culture into the culture medium, and then seal the bottle as it is. After inoculation, place it in an environment with a temperature of 24°C, a relative humidity of 50%, and good ventilation for 45 days. Take out the grown culture, stir it evenly, and then dry it immediately at 70°C to obtain dried culture.

[0026] Example 2 Preparation of Cs-2025 culture The formula of the culture medium is: millet 180 g, sucrose 8 g, magnesium sulfate 200 mg, potassium dihydrogen phosphate 400 mg, and distilled water 400 mL.

[0027] Preparation of culture medium: Dissolve all ingredients except millet in water, adjust pH to 6.0, add rice, continue to heat and stir, stop heating when it becomes solid, fill into 10 500ml jars, seal with a layer of cellophane paper, sterilize in a high pressure steam sterilizer at 121°C for 20 minutes, cool to below 27°C, and store for use.

[0028] Inoculation and culture: Open the jar under sterile conditions, inoculate with mother culture, seal the jar, and place in a 22°C, 30% relative humidity, well ventilated environment for 45 days. Take out the culture, mix well, and dry at 60°C to obtain dried culture.

[0029] Example 3 Preparation of Cs-2025 culture Culture medium: millet 220g, sucrose 12g, magnesium sulfate 300mg, potassium dihydrogen phosphate 600mg, distilled water 600ml.

[0030] Preparation of culture medium: Dissolve all ingredients except millet in water, adjust pH to 7.0, add rice, continue to heat and stir, stop heating when it becomes solid, fill into 10 500ml jars, seal with a layer of cellophane paper, sterilize in a high pressure steam sterilizer at 121°C for 20 minutes, cool to below 27°C, and store for use.

[0031] Inoculation and culture: Open the jar under sterile conditions, inoculate with mother culture, seal the jar, and place in a 26°C, 60% relative humidity, well ventilated environment for 45 days. Take out the culture, mix well, and dry at 80°C to obtain dried culture.

[0032] Example 4 Therapeutic effect of Cs-2025 culture on kidney function damage in mice induced by adenine The Cs-2025 culture used in this example was obtained by the method described in Example 1. Kunming mice, male, weight 22-26g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. Adenine was purchased from BIO. BASIC. INC.

[0033] The traditional method of preparing animal kidney function damage model is mostly 5 / 6 nephrectomy to model, but there are many shortcomings such as complicated operation, difficult to make, many complications, high mortality and unable to prepare different degrees of kidney function damage model. In this study, the method of continuous gavage of high concentration of adenine was used to prepare mouse kidney function damage model. High concentration of adenine is changed into 2, 8-dihydroxyadenine which is extremely difficult to dissolve in water by the action of xanthine oxidase, which deposits in the renal tubules, affects the excretion of nitrogen compounds, and ultimately causes kidney damage. In addition, the large intake of adenine leads to excessive production of oxygen free radicals and lipid peroxidation in the body, which may be an important mechanism of adenine leading to kidney damage.

[0034] Animal grouping and administration method: 50 male Kunming mice were randomly divided into normal control group, model control group, Cs-2025 high dose group, Cs-2025 medium dose group and Cs-2025 low dose group. Except for the normal control group, the other groups of mice were given adenine 250mg / kg by gavage on the 1st, 3rd and 5th day, and then supplemented every 10 days. After the first three times of adenine administration, each dose group was given Cs-2025 by gavage for 30 days, and the gavage amount was 1000mg / kg, 500mg / kg and 250mg / kg respectively; the normal control group was given 1mL / 100g distilled water every day.

[0035] General observation: During the experiment, the changes of mouse fur luster, mental state, food and water intake, urine volume and body weight were observed.

[0036] Blood biochemical index: After the administration was completed, the mice were weighed, the mice were anesthetized by intraperitoneal injection of sodium pentobarbital (60mg / kg), the eyeball was taken to collect blood, the serum was separated, and the serum BUN and CRE contents were determined by automatic biochemical analyzer (Mindray).

[0037] Organ index and kidney pathology: The left kidney, right kidney, spleen and thymus were weighed respectively; the left kidney was stripped and fixed with 10% formaldehyde overnight, paraffin-embedded, HE stained, and the pathological changes of kidney were observed.

[0038] Statistical analysis: The experimental data were expressed as , and the statistical software was used for data statistical analysis, the mean comparison between groups was analyzed by one-way analysis of variance, the comparison between two groups was analyzed by least significant difference method (LSD), and the test level was taken as α=0.05.

[0039] (1) The general observation results of animals in each group Compared with the normal control group, the model control group mice were less active, with loose and dull fur, reduced food intake, significantly increased water and urine volume, and significantly decreased body weight. The body weight change was statistically different compared with the normal control group (see Table 1). The Cs-2025 administration groups were significantly improved compared with the model control group, with increased activity, slightly shiny fur, less increased urine volume, and less decreased body weight (see Table 1). The body weight change was statistically different compared with the model group P <0.05).

[0040] Table 1. Effect of Cs-2025 on body weight of mice with renal function injury

[0041] Note: * indicates a significant difference compared with the normal control group P <0.05); △ indicates a significant difference compared with the model group P <0.05).

[0042] (2) Effect of Cs-2025 on serum BUN and CRE levels of mice with renal function injury Compared with the normal control group, the serum BUN and CRE of the model control group mice were significantly increased P <0.05); Compared with the model control group, the serum BUN and CRE levels of the Cs-2025 dose groups were significantly decreased P <0.05) (see Table 2), indicating that Cs-2025 had a good improvement effect on adenine-induced renal function changes.

[0043] Table 2. Effect of Cs-2025 on serum CRE and BUN levels of mice with renal function injury

[0044] Note: * indicates a significant difference compared with the normal control group P <0.05); △ indicates a significant difference compared with the model group P <0.05).

[0045] (3) Effect of Cs-2025 on organ index and kidney pathology of mice with renal function injury The kidney size of the normal control group mice was moderate, the color was brownish red, and the surface was smooth, while the kidney of the model group animals was significantly enlarged, and the kidney index increased P< 0.01) (see Table 3), pale, surface uneven, the kidney color of Cs-2025 each administration group was reddish-brown, surface smooth, and the kidney index of Cs-2025 each administration group was obviously reduced P < 0.01) (see Table 3).

[0046] Compared with the normal control group, the thymus of the model group was significantly atrophic and small, the spleen was obviously small, the thymus index and the kidney index were reduced P < 0.01) (see Table 3), and the thymus and the spleen of Cs-2025 each administration group were close to the normal control group, the thymus index and the kidney index were increased compared with the model control group, and were close to the normal control group P < 0.01) (see Table 3).

[0047] Table 3. Effect of Cs-2025 on the organ index of mice with renal function injury

[0048] Note: * Significant difference compared with the normal control group P < 0.05; △ Significant difference compared with the model group P < 0.05.

[0049] From the pathological results, it can be seen that the cortex and medulla of the kidney of the normal control group mouse were clear, the number and shape of glomerulus were normal, and the shape of renal tubule was normal: no expansion, intact epithelium, and no edema phenomenon Figure 1 ); the renal tubule of the model mouse was severely expanded, the renal tubule epithelium was edematous, and the renal interstitium was infiltrated with inflammatory cells Figure 2 ); the renal tubule of the Cs-2025 administration group mouse was slightly expanded, and no epithelial edema was observed, and the inflammatory cell infiltration of the renal interstitium was reduced Figure 3 .

[0050] Through the experiment, we confirmed that the culture of Cs-2025 strain has a good protective effect on the renal function injury caused by adenine.

[0051] The above only is the specific implementation of the present application, enables the person skilled in the art to understand or implement the present application. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present application. Accordingly, the present application will not be limited to these embodiments described herein but will be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. Use of a fungal strain Cs-2025 in the preparation of a drug for treating impairment of renal function, the strain Cs-2025 being deposited with the China General Microbiological Culture Collection Center and having the accession number CGMCC NO. 41879.

2. A medicament for treating impairment of renal function, characterized by comprising: A culture comprising the fungal strain Cs-2025 of claim 1.

3. The medicament according to claim 2, characterized in that: The drug treats impairment of renal function by reducing purine levels.

4. A method for preparing a culture of a fungal strain Cs-2025, characterized by, The mother culture of the strain Cs-2025 is inoculated into a culture medium and cultured, and after the culture is completed, the culture is dried to obtain a culture.

5. The method of claim 4, wherein: The culture medium comprises, in mass fractions: millet 180-220 g, sucrose 8-12 g, magnesium sulfate 200-300 mg, potassium dihydrogen phosphate 400-600 mg, and distilled water 400-600 ml.

6. The method of claim 4, wherein: The culture medium comprises, in mass fractions: millet 200 g, sucrose 10 g, magnesium sulfate 250 mg, potassium dihydrogen phosphate 500 mg, and distilled water 500 ml.

7. The production method according to claim 4 or 5, characterized by: The pH value of the culture medium is 6.0-7.

0.

8. The method of claim 7, wherein: The temperature during the culture is controlled at 22-26°C.

9. The method of claim 8, wherein: The relative humidity during the culture is 30-80%.

10. The method of claim 9, wherein: Freeze-drying or drying at a temperature of 60-80°C.

Images