Application of rhodotorula red in preparation of medicine for treating diabetic nephropathy

Abstract

The invention provides application of rhodotorula red in preparation of a medicine for treating diabetic nephropathy, and relates to the technical field of biological medicine. According to the application, by constructing a mouse diabetic nephropathy model, the control and alleviation effects of the rhodotorula red pigment on the diabetic kidney diseases are detected on an animal level, and particularly by observing indexes such as blood sugar, renal functions, renal tissue injury conditions and the like of diabetic mice and structures of intestinal microbial flora and the like of the rhodotorula red pigment, the diabetic kidney diseases can be controlled and relieved. The important function of the rhodotorula red in controlling and relieving the diabetic nephropathy is proved; besides, as a food source compound with high safety, the rhodotorula rubra can be used for relieving and treating kidney diseases of diabetes mellitus by feeding the rhodotorula rubra, so that an effective component with safety is provided for daily health care of diabetic nephropathy; theoretical and experimental basis is provided for development of daily health-care components of the rhodotorula red pigment for auxiliary treatment of diabetes mellitus.

Description

Technical Field

[0001] The present application relates to the field of biomedicine technology, and in particular to the use of rhodotorula erythrosin in the preparation of a drug for treating diabetic nephropathy. Background Art

[0002] Diabetic kidney disease (DKD) is the most common microvascular complication of diabetes and has become the main cause of chronic kidney disease and end-stage renal disease in China. Current methods for treating diabetic nephropathy include blood sugar or blood pressure control, but existing drugs have many application restrictions on the patient's age, health status, etc., and most drugs have strong toxic side effects. In recent years, the importance of the intestinal flora-gut kidney axis in the pathogenesis of diabetic nephropathy has gradually attracted attention. Intestinal flora imbalance can further damage the kidneys by affecting metabolites, inflammatory factors and other pathways. There are many application restrictions on existing drugs, and many drugs currently used to treat diabetic nephropathy have many application restrictions in actual application. For example, sulfonylurea drugs should be prohibited or used with caution in specific populations such as children under 10 years old, the elderly over 80 years old, pregnant and lactating women, and those with liver and kidney dysfunction. Blood pressure control drugs should be prohibited in cases of pregnancy, bilateral renal artery stenosis, acute heart failure, sick sinus syndrome, etc., or allergies to drugs. When using these drugs, doctors need to make individual adjustments based on the specific conditions of the patients. Blood sugar control drugs may cause hypoglycemia reactions, such as dizziness, fatigue, sweating, etc., and in severe cases, coma, and other adverse reactions, such as nausea, vomiting, diarrhea, etc. Lipid-lowering drugs (atorvastatin, rosuvastatin, etc.) and RAS blockers (sartans, prils) may cause adverse reactions such as liver damage and muscle pain. In addition, a small number of patients will have allergic reactions, and anaphylactic shock is occasionally seen. When using these drugs, doctors need to closely monitor the patient's condition changes and adverse reactions. In addition, long-term medication can lead to a disorder in the structure of intestinal microbial flora, thereby causing changes in intestinal metabolites and inducing other side effects and complications. These reasons all bring certain difficulties to the daily medication selection and dosage of more sensitive people with diabetes or diabetes-related diseases. Therefore, it is urgent to develop a health care functional component for auxiliary treatment with high safety and diabetes and complication control effects.

[0003] Rhodotorula hexogenin is a new type of natural carotenoid. In past studies, the extremely superior anti-inflammatory and anti-tumor properties of rhodotorula hexogenin are irreplaceable by traditional carotenoids. However, most of the existing research on the function of rhodotorula hexogenin is focused on the cellular level, and there is very little research at the animal level. In addition, there is no related application of rhodotorula hexogenin in the field of alleviating diabetic nephropathy. Summary of the invention

[0004] The purpose of the present application is to provide the use of rhodotorula erythrophylla in the preparation of a drug for treating diabetic nephropathy and to expand the application scope of rhodotorula erythrophylla.

[0005] In order to solve the above technical problems, the technical solution adopted in this application is:

[0006] The present application provides the use of rhodotorula erythrophyllotoxin in the preparation of a drug for treating diabetic nephropathy. The concentration of rhodotorula erythrophyllotoxin in the drug is 3 mg / mL, and the dosage of rhodotorula erythrophyllotoxin is 30 mg / kg.

[0007] Although relevant studies have reported that red yeast rhodopsin can relieve hyperlipidemia chronic kidney disease by regulating oxidative stress levels, hyperlipidemia chronic kidney disease is mainly caused by abnormal lipid metabolism, which leads to decreased kidney metabolism of lipoproteins and increased lipoprotein levels in the blood, while diabetic nephropathy is caused by long-term hyperglycemia and insulin resistance. Hyperglycemia can cause microvascular damage in the kidneys, impairing glomerular filtration function and thus causing proteinuria. In addition, hyperglycemia can also promote oxidative stress response, accelerate renal cell apoptosis, and further aggravate kidney damage. Therefore, there is a significant gap between the pathogenesis of hyperlipidemia chronic kidney disease and diabetic nephropathy described in this application, and the animal level disease model used is also different. This application has found that feeding red yeast rhodopsin can reduce the blood sugar and oxidative stress levels of diabetic nephropathy mice to a certain extent, and repairs the disordered microbial flora structure caused by diabetes, effectively alleviating kidney tissue damage in diabetic nephropathy mice, which is different from the principle mechanism of hyperlipidemia chronic kidney disease.

[0008] Compared with the prior art, the embodiments of the present application have at least the following advantages or beneficial effects:

[0009] The present application constructs a diabetic nephropathy model in mice to detect the control and relief effects of rhodotorula rhodotorula on diabetic nephropathy at the animal level. Specifically, the effects of rhodotorula rhodotorula on blood sugar, renal function, renal tissue damage, and the structure of intestinal microbial flora in diabetic mice are observed to prove the important function of rhodotorula rhodotorula in controlling and relieving diabetic nephropathy. In addition, rhodotorula rhodotorula is a highly safe food-derived compound. Feeding rhodotorula rhodotorula can relieve and treat diabetic kidney disease, providing a safe and effective ingredient for the daily health care of diabetic nephropathy, and providing a theoretical and experimental basis for the development of rhodotorula rhodotorula as a daily health care ingredient for the auxiliary treatment of diabetes. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0011] Figure 1 This is a comparison chart of the body weight and fasting blood glucose of mice in different experimental groups in the present application example;

[0012] Figure 2 This is a comparison chart of the 24-hour food intake, water intake, and urine output of mice in different groups in the present application example;

[0013] Figure 3 This is a comparison chart of the levels of serum creatinine in blood tissue, urine creatinine in urine tissue, and urine albumin in different groups of mice in the examples of this application;

[0014] Figure 4 The following are comparison diagrams of glomerular structures in kidney tissues of mice in different groups under transmission electron microscopy (observation magnifications are 3000×, 7000×, and 20000×) in the examples of the present application;

[0015] Figure 5 This is a comparison of the H&E, PAS and Masson staining results in the kidney tissues of different groups of mice in the examples of this application;

[0016] Figure 6 This is a comparison chart of the expression levels of Nrf2, keap1, HO-1, NQO1 and β-actin proteins in the kidney tissues of different groups of mice in the examples of this application;

[0017] Figure 7 This is a comparison chart of the expression levels of malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (CAT and SOD) in the blood of different groups of mice in the examples of this application;

[0018] Figure 8 This is the PLS-DA analysis diagram of the DKD group and the DKD+Toru group in the examples of this application;

[0019] Fig. 9 This is a diagram showing the effect of red yeast rhodotorula on the intestinal microbial structure at the phylum level in the example of this application;

[0020] Fig.10 This is a correlation analysis diagram of intestinal microorganisms and metabolites based on Mantel-test in the embodiment of the present application. DETAILED DESCRIPTION

[0021] In order to make the purpose, technical scheme and advantages of the embodiments of the present application clearer, the technical scheme in the embodiments of the present application will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to the conventional conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0022] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present application can be combined with each other. The present application will be described in detail below with reference to specific embodiments.

[0023] The present application discloses the use of rhodotorula erythrophyllin in the preparation of a drug for treating diabetic nephropathy, wherein the chemical structural formula of rhodotorula erythrophyllin is as shown in Formula I:

[0024]

[0025] In some embodiments of the present application, the above-mentioned drug further comprises a pharmaceutically acceptable carrier.

[0026] In some embodiments of the present application, the above-mentioned drug is in oral dosage form.

[0027] In some embodiments of the present application, the above-mentioned drugs are used to repair the intestinal microbial flora structure.

[0028] This application uses a mouse model to explore the effect and mechanism of red yeast rhodotorula in treating diabetic nephropathy. The process is as follows:

[0029] (1) Construction and grouping of diabetic mouse models: ① Normal non-modeled control group (Control, Con); ② Diabetic nephropathy model group (DKD); ③ Model mice + administration of red yeast rhodotorula group (DKD+Toru); ④ Model mice + administration of dapagliflozin group (DKD+Dapa);

[0030] (2) Administration: The dosage of rhodotorula rubrum was 30 mg / kg (dissolved in corn oil, drug concentration 3 mg / mL) and the administration was continued for 2 months;

[0031] (3) Record the blood glucose and body weight of mice;

[0032] (4) Sampling: Collect the metabolic status of each group of mice for 24 hours, dissect the mice, and separate and collect the blood, urine, and kidney tissues of the mice;

[0033] (5) Detection of indicators related to diabetic kidney tissue damage;

[0034] (6) Detect oxidative stress levels in blood and kidney tissue;

[0035] (7) Analyze the structure of intestinal microbial flora;

[0036] (8) Evaluate the protective and auxiliary therapeutic effects of red yeast rhodotorula on diabetic kidney damage.

[0037] The features and performance of the present application are further described in detail below in conjunction with the embodiments.

[0038] Example

[0039] 1. Construction of diabetic mouse model

[0040] The diabetic model of mice was induced by intraperitoneal injection of streptozotocin (STZ, purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.). Fifty 8-week-old male mice with C57BL / 6 background (purchased from Wuhan Shubaili Biotechnology Co., Ltd.) were purchased and intraperitoneally injected with STZ (50 mg / kg, dissolved in sodium citrate buffer) for 5 consecutive days, and fed with a high-fat diet. At the same time, a control group of mice without modeling was set up (the same volume of sodium citrate buffer was injected intraperitoneally and fed with ordinary maintenance feed). One week later, the mice were fasted and deprived of water for 12 hours, and the fasting blood glucose (FBG) was >11.1mmol / L, which was considered as diabetic mice; those below 11.1mmol / L were considered as healthy mice without modeling. The mice were divided into groups (n=6): ① Ordinary non-modeling control group (Control, Con); ② Diabetic nephropathy modeling group (DKD); ③ Modeling mice + administration of red yeast rhodotorula group (DKD+Toru); ④ Modeling mice + administration of dapagliflozin group (DKD+Dapa).

[0041] 2. Drug administration

[0042] One month after the diabetic mice were established, the mice in the ③DKD+Toru group were given rhodotorula rhodotorula (30 mg / kg, dissolved in corn oil, drug concentration 3 mg / mL) by oral gavage, and the mice in the ④DKD+Dapa group were given dapagliflozin (2 mg / kg, turbid solution prepared with sterile water, drug concentration 0.2 mg / mL, as a positive control) by oral gavage. The drugs were continued for 2 months.

[0043] 3. Record blood glucose and body weight of mice

[0044] During the modeling and drug administration period, the body weight and fasting blood glucose (FBG) of the mice were measured and recorded every two weeks until the end of the experiment. The comparison of body weight and fasting blood glucose of mice in different groups is shown in the figure below. Figure 1 As shown, Figure 1 The left is a comparison chart of body weight. Figure 1 The right is a comparison chart of fasting blood glucose; Figure 1 The effects of rhodotorula on body weight and blood sugar in a diabetic model can be seen in:

[0045] ① Body weight loss was reduced: Compared with healthy mice without modeling (Con group: 25.05±0.76g), diabetic mice (DKD group: 18.12±1.50g) had significantly reduced body weight. After feeding diabetic mice with rhodozyma (DKD+Toru group: 22.97±1.10g), the weight loss of mice was reduced, which is also a positive effect for the treatment of diabetes. Feeding dapagliflozin (DKD+Dapa: 22.3±1.93g) can also slow down the weight loss of mice.

[0046] ② Decreased fasting blood glucose (FBG) levels: Increased blood glucose levels are the most significant symptom of diabetes. Compared with healthy mice without modeling (Con group: 4.80±0.77mmol / L), fasting blood glucose levels in diabetic mice (DKD group: 15.40±5.21mmol / L) were significantly increased. After feeding diabetic mice with rhodozyma (DKD+Toru group: 4.98±1.05mmol / L), the glucose level in the blood of diabetic mice was significantly reduced, thereby reducing the complications caused by high sugar to the body. Feeding dapagliflozin (DKD+Dapa: 5.20±0.8mmol / L) can also reduce fasting blood glucose levels.

[0047] 4. Sampling

[0048] After 2 months of administration, the 24-hour food intake, water intake, and urine volume of the mice were collected and recorded using metabolic cages, urine was collected, blood was collected through orbital bleeding, and the mice were dissected to separate and collect the kidney tissue of the mice for subsequent testing. Figure 2 shown.

[0049] 24h metabolic indicators returned to normal: During the development of diabetes, "three mores and one less" will occur, that is, compared with normal mice, diabetic mice will have more 24-hour food intake (Con group: 2.50±0.6g; DKD group: 4.24±1.14g), water intake (Con group: 2.50±0.55mL; DKD group: 5.67±1.21mL) and urine volume (Con group: 0.67±0.30mL; DKD group: 2.28±0.31mL). Figure 2It can be seen that after diabetic mice were fed with rhodotorula rhodotorula, the 24-hour food intake (DKD+Toru group: 2.58±0.5g), water intake (DKD+Toru group: 4.00±0.63mL) and urine volume (DKD+Toru group: 1.75±0.21mL) of the mice returned to normal levels, reducing the pressure on the kidneys to a certain extent. After administration of dapagliflozin, the 24-hour food intake (DKD+Dapa group: 2.35±0.47g), water intake (DKD+Dapa group: 3.00±0.89mL) and urine volume (DKD+Dapa group: 0.92±0.52mL) of the mice also returned to normal levels.

[0050] 5. Detection of physiological and biochemical indicators related to kidney disease in blood / urine tissue samples

[0051] Biochemical kits (purchased from Wuhan Elabscience Biotechnology Co., Ltd.) were used to measure the physiological and biochemical indicators of diabetic nephropathy in the blood: urea nitrogen (BUN); and the physiological and biochemical indicators of diabetic nephropathy in the urine: urinary creatinine levels and urinary albumin levels. The comparison of serum creatinine in blood tissues, urinary creatinine in urine tissues and urinary albumin levels in different groups of mice was as follows: Figure 3 shown.

[0052] from Figure 3 It can be seen that the effect of red yeast rhodotorula on renal function in diabetic nephropathy:

[0053] ① Physiological indicators of kidney disease in blood tissues: Long-term high blood sugar environment can cause damage to the kidneys, leading to a decrease in glomerular filtration rate and a decrease in the kidney's ability to excrete urea nitrogen, which in turn causes an increase in the level of urea nitrogen (BUN) in the blood. Compared with healthy mice without modeling (Con group: 8.43±1.33mmol / L), the level of urea nitrogen in the blood of diabetic mice (DKD group: 11.17±1.38mmol / L) was significantly increased. After diabetic mice were fed with red yeast rhodopsin (DKD+Toru group: 9.10±0.63mmol / L), the level of urea nitrogen in the blood of mice was reduced. Feeding dapagliflozin (DKD+Dapa: 9.08±0.88mmol / L) can also reduce the level of urea nitrogen in the blood.

[0054] ② Kidney pathology-related indicators in urine tissue: Changes in urine creatinine and albumin levels are usually related to the filtration function and urine concentration ability of the kidney. Compared with normal mice, diabetic mice had increased levels of urine creatinine (Con group: 1580.16±117.24mmol / L; DKD group: 1836.28±119.23mmol / L) and urine albumin (Con group: 12.39±0.53ng / mL; DKD group: 14.75±0.18ng / mL). After diabetic mice were fed with rhodozyma, the levels of urine creatinine (DKD+Toru group: 1688.17±71.70mmol / L) and urine albumin (DKD+Toru group: 13.86±0.64ng / mL) in the mice's urine were downregulated and restored to normal levels, indicating that rhodozyma can improve the filtration function of diabetic kidney tissue. After administration of dapagliflozin, the urinary creatinine (DKD+Dapa group: 1606.06±75.18mmol / L) and urinary albumin (DKD+Dapa group: 13.77±0.36ng / mL) in the urine of mice also returned to normal levels.

[0055] 6. Pathological testing of kidney tissue samples

[0056] After the kidney cortex was dissected, it was placed in electron microscopy fixative (purchased from Servicebio) at room temperature in the dark for 2 hours, then transferred to 4°C for storage, and the samples were sent to Servicebio for transmission electron microscopy and photography, and the structure of the glomeruli in the kidney cortex area was recorded at observation magnifications of 3000×, 7000×, and 20000×. Comparison of glomerular structures under transmission electron microscopy (observation magnifications of 3000×, 7000×, and 20000×) in the kidney tissues of different groups of mice Figure 4 shown.

[0057] The integrity of the glomerular structure in the renal cortex, the normal structure of podocytes and basement membranes are the basis for the normal physiological function of the kidney. Figure 4 It can be seen that compared with the control group (Con), the foot processes of glomerular podocytes in the diabetic nephropathy model group (DKD) fused and disappeared, the basement membrane thickened unevenly, and the matrix concentration decreased. After feeding red yeast rhodotorula (DKD+Toru), the foot process fusion improved, the uneven basement membrane thickening phenomenon was alleviated, and the matrix concentration increased. This shows that red yeast rhodotorula effectively improves the degree of damage to the glomeruli in the cortical area of ​​the kidney in diabetes.

[0058] After the kidney tissue was dissected, it was placed in a fixative (purchased from Servicebio) and fixed at room temperature for 24 hours. The samples were sent to Servicebio for H&E, PAS and Masson staining and scanning and photography to observe the pathological changes of the kidney tissue. Figure 5 shown.

[0059] from Figure 5 It can be seen that the effect of red yeast rhodotorula on renal pathological damage in diabetic nephropathy:

[0060] ①H&E staining: The renal structure of mice in the control group (Con) was clearly visible and intact. The renal tubules were regular in shape and there was no inflammatory cell infiltration. Compared with the control group (Con), the diabetic nephropathy model group (DKD) had thickened basement membrane, mesangial cell proliferation, obvious tubular dilatation, and glomerular hypertrophy. Compared with the DKD group, after feeding red yeast rhodopsin (DKD+Toru), the lesions were significantly improved, the basement membrane thickening and mesangial cell proliferation were improved, the renal tubules were more complete, and the glomerular hypertrophy was reduced.

[0061] ②PAS staining: In the kidney tissue of mice in the control group (Con), the positive areas were mainly distributed along the glomerular capillary wall and renal tubules, with no positive areas in the middle. Compared with the control group (Con), the model group (DKD) had significantly more positive substance deposition, thickened basement membrane, expanded mesangial area, and increased glycogen accumulation in the cytoplasm. Compared with the DKD group, after feeding red yeast rhodopsin (DKD+Toru), the positive area was significantly reduced, the renal tubular structure was relatively regular, the positive substance deposition phenomenon was reduced, and the basement membrane thickening and mesangial hyperplasia phenomenon were improved.

[0062] ③Masson staining: The kidneys of mice in the control group (Con) were all red-stained normal tissues. Compared with the control group (Con), a large number of blue fibers were visible in the kidney tissue of the model group (DKD). Compared with the DKD group, the blue fibers were significantly reduced after feeding red yeast rhodopsin (DKD+Toru).

[0063] Therefore, the staining results show that red yeast rhodotorula can significantly improve the degree of renal pathological damage in diabetic nephropathy, slow down the thickening of basement membrane and mesangial hyperplasia, and reduce glycogen accumulation and fibrosis.

[0064] 7. Detection of oxidative stress levels in diabetic nephropathy

[0065] Immunoblotting was used to detect the activation level of Nrf2-Keap1-mediated oxidative stress signaling pathway in different groups of kidney tissues. RIPA lysis buffer (purchased from Beyotime Biotechnology Co., Ltd.) was added to the kidney tissue to prepare tissue homogenate, and 5×SDS loading (purchased from Wuhan Servicebio Biotechnology Co., Ltd.) was added after centrifugation to obtain the supernatant. The protein was denatured by heating, and 10% SDS polyacrylamide gel was prepared to ensure that an equal amount of protein sample was added to each group. Electrophoresis was performed in an electrophoresis instrument for 1-2 hours, and then the protein sample was transferred to a PVDF membrane by a membrane transfer instrument according to the protein molecular weight. After the transfer was completed, the membrane was cut according to the molecular weight, and 5% skim milk was added as a blocking solution for 1 hour at room temperature. The membrane was incubated with appropriately diluted specific Nrf2, keap1, HO-1, NQO1 and β-Actin antibodies at 4°C overnight. The membrane was then washed 6 times with TBST solution, incubated with the corresponding secondary antibody for 2 hours, and then washed 6 times with TBST solution. Finally, ECL was used for color development and observed with a chemiluminescence analyzer. (All antibodies were purchased from Hangzhou Huaan Biotechnology Co., Ltd.: HUABIO Biotechnology Co., Ltd., and the remaining reagents and consumables were purchased from Wuhan Servicebio Biotechnology Co., Ltd.: Servicebio Biotechnology Co., Ltd.) Comparison of Nrf2, keap1, HO-1, NQO1 and β-actin protein expression levels in kidney tissues of mice in different groups Figure 6 shown.

[0066] Biochemical kits (purchased from Wuhan Elabscience Biotechnology Co., Ltd.) were used to measure the oxidative stress-related indicators malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (CAT and SOD) in the blood. Comparison of the expression levels of malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (CAT and SOD) in the blood of mice in different groups Figure 7 shown.

[0067] from Figure 6 and Figure 7 As can be seen in the results, the effects of rhodotorula on oxidative stress levels in diabetes include:

[0068] ①Nrf2-Keap1-mediated oxidative stress signaling pathway: Under normal physiological conditions, NRF2 binds to KEAP1 and is ubiquitinated and degraded through the ubiquitin-proteasome system, thereby maintaining a low level of NRF2. When the oxidation level increases, the levels of oxidants such as reactive oxygen species (ROS) and reactive nitrogen species (RNS) in the cell increase, causing the conformation of KEAP1 to change, and then dissociate from NRF2. The dissociated NRF2 is activated and translocated from the cytoplasm to the nucleus, promoting the level of oxidative stress. When the oxidation level in the tissue is upregulated, the protein levels of NRF2 and KEAP1 increase, and the protein levels of the antioxidant enzymes HO-1 and NQO1 are downregulated. The immunoblotting experiment showed that compared with the control group (Con), the diabetic nephropathy model group (DKD) had increased Nrf2 and Keap1 protein levels, and decreased HO-1 and NQO1 protein levels. After feeding red yeast rhodotorula (DKD+Toru), the Nrf2 and Keap1 protein levels were significantly reduced, and the protein levels of antioxidant enzymes HO-1 and NQO1 were promoted. This indicates that red yeast rhodotorula can regulate the Nrf2-Keap1-mediated oxidative stress signaling pathway, promote antioxidant levels, and alleviate the oxidation level in renal tissue, thereby playing a protective auxiliary therapeutic role in diabetic nephropathy.

[0069] ② Decreased oxidation level: Malondialdehyde (MDA) is one of the final products of lipid peroxidation in organisms and is often used to assess the degree of oxidative damage to cells or tissues. Compared with healthy mice without modeling (Con group: 9.96±6.73mol / L), the MDA level in the blood of diabetic mice (DKD group: 23.88±6.00mol / L) was significantly increased. After diabetic mice were fed with rhodotorula rhodotorula (DKD+Toru group: 10.96±4.81mol / L), the MDA level in the blood of mice was reduced. After feeding with dapagliflozin (DKD+Dapa: 12.29±5.98mol / L), the MDA level in the blood was also reduced, indicating that the oxidation level in the body was reduced.

[0070] ③ Upregulation of antioxidant levels: Reduced glutathione (GSH) and superoxide dismutase (CAT and SOD) are often used to evaluate the antioxidant capacity of tissues. Compared with normal mice, the levels of GSH (Con group: 4238.62.16±90.23U; DKD group: 3870.63±116.40U), CAT (Con group: 134.13±17.35U / mL; DKD group: 88.95±2.39U / mL), and SOD (Con group: 126.60±10.81U; DKD group: 102.35±11.38U) in the blood of diabetic mice were reduced. After diabetic mice were fed with rhodotorula rhodotorula, the levels of GSH (DKD+Toru group: 4455.08±372.43U), CAT (DKD+Toru group: 124.72±6.72U / mL), and SOD (DKD+Toru group: 142.42±14.20U) in the blood of mice increased. Compared with the DKD group, the levels of GSH (DKD+Dapa group: 4449.49±269.35U), CAT (DKD+Dapa group: 123.52±10.39U / mL), and SOD (DKD+Toru group: 170.67±13.11U) in the blood of mice also increased after administration of dapagliflozin. The upregulation of reduced glutathione and superoxide dismutase levels in the blood indicates that the body's antioxidant level is increased, which helps to reduce the level of oxidative stress in diabetic kidneys, thereby improving the protective effect on diabetic kidney tissue.

[0071] 8. The structural composition of intestinal microorganisms in diabetic nephropathy

[0072] The feces of mice in the ②DKD group and ③DKD+Toru group were collected, stored in dry ice and sent to Shanghai Meiji Biotechnology Co., Ltd. for microbiome analysis. The genome was extracted and the 16S sequence was sequenced to analyze the effect of red yeast rhodotorula on the structure of the intestinal microbial flora of mice.

[0073] All data analyses were performed on the MajorBio cloud platform (https: / / cloud.majorbio.com), as follows: mothur software (http: / / www.mothur.org / wiki / Calculators) was used to calculate the Alpha diversity knowledge Chao 1, Shannon index, etc., and the Wilxocon rank sum test was used to analyze the inter-group differences in Alpha diversity: PCoA analysis (principal coordinate analysis) based on the Bray-Curtis distance algorithm was used to test the similarity of the microbial community structure between samples, and the PERMANOVA non-parametric test was combined to analyze whether the differences in microbial community structure between sample groups were significant; LEfSe analysis (Linear discriminant analysis Effect Size) (http: / / huttenhower.sph.harvard.edu / LEfSe) (LDA>2, P<0.05) was used to determine the bacterial groups with significant differences in abundance from the phylum to the genus level between different groups. Distance-based redundancy analysis (db-RDA) was used to investigate the influence of metabolic indicators on the intestinal bacterial community structure. Linear regression analysis was used to evaluate the effects of the main soil physicochemical / clinical indicators determined in the db-RDA analysis on the microbial Alpha diversity index. Species were selected for correlation network analysis based on spearman correlation |r|>0.6p<0.05. Correlation analysis based on Mantel-test was used to determine the correlation between microorganisms and metabolites.

[0074] From the analysis results, we can see that the intra-group parallelism of this microbiome study is good, and the differences between groups are significant ( Figure 8 ), the intestinal microbial structure of the DKD group and the DKD+Toru group was significantly different, which also means that the intake of rhodotorula rhodotorula did change the structure of the intestinal flora of mice. From the statistical results of the bacterial phyla of the two groups, Firmicutes, Bacteroidetes and Desulfobacteria were the main components of the intestinal flora of the two groups of mice. Rhodotorula rhodotorula significantly increased the abundance of Bacteroidetes and reduced the abundance of Firmicutes and Desulfobacteria ( Fig. 9 ). The Bacteroidetes flora and carbohydrate metabolism are closely related. In addition, this application screened a variety of significantly different metabolites in the intestine and conducted correlation analysis with significantly different bacterial genera, and found that changes in the microbial flora are indeed closely related to some metabolites closely related to diabetic nephropathy ( Fig.10 ).

[0075] In summary, the examples of this application demonstrate that rhodotorula erythrin has important application value in the process of alleviating and treating diabetic kidney disease. This patent constructs a mouse diabetic nephropathy model and detects the control and relief effect of rhodotorula erythrin on diabetic kidney disease at the animal level. It provides a daily health care ingredient and theoretical basis with higher safety for the control and auxiliary treatment of diabetic nephropathy.

[0076] The embodiments described above are part of the embodiments of the present application, rather than all of the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents the selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work belong to the scope of protection of the present application.

Claims

1. Use of red yeast rhodotorula in the preparation of drugs for treating diabetic nephropathy.

2. The use of rhodotorula erythrin according to claim 1 in preparing a drug for treating diabetic nephropathy, characterized in that: The medicament further comprises a pharmaceutically acceptable carrier.

3. The use of rhodotorula erythrin in the preparation of a drug for treating diabetic nephropathy according to claim 2, characterized in that: The drug is in oral dosage form.

4. The use of rhodotorula erythrin in the preparation of a drug for treating diabetic nephropathy according to claim 1, characterized in that: The medicine is used to repair the intestinal microbial flora structure.

5. The use of rhodotorula erythrin in the preparation of a drug for treating diabetic nephropathy according to claim 1, characterized in that: The concentration of rhodotorula rhodotorula in the medicine is 3 mg / mL.

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