Traditional Chinese medicine composition for preventing and treating UACR rise and application

Abstract

The invention belongs to the field of traditional Chinese medicine, and discloses a traditional Chinese medicine composition for preventing and treating UACR rise and application. The traditional Chinese medicine composition is prepared from the following components in parts by mass: 40-60 parts of radix astragali, 25-35 parts of semen euryales, 18-22 parts of fructus rosae laevigatae, 4-6 parts of leeches, 18-22 parts of caulis sinomenii and 35-55 parts of penthorum chinense pursh. The traditional Chinese medicine composition has the effects of tonifying spleen and kidney and reducing turbid toxins, and has the effect of protecting the liver, so that the safety is relatively high; moreover, the treatment effect on the reduction or reversal of the urine UACR is better, the recovery is faster, and the application prospect in the field of treatment of the diabetic nephropathy at the middle and early stage is wide.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine, and in particular relates to a traditional Chinese medicine composition for preventing and treating elevated UACR and its application. Background Art

[0002] Diabetic kidney disease (DKD) is a diabetic complication characterized by renal microvascular lesions. About 30%-40% of diabetic patients progress to DKD, which is the primary cause of end-stage renal disease (ESRD). Modern medicine uses multiple risk factor interventions, such as blood pressure reduction, blood sugar and lipid regulation, and renin-angiotensin system inhibitors (RASi), but it is still difficult to inhibit the progression of DKD. Evidence-based evidence shows that sodium-glucose cotransporter 2 inhibitors (SGLT2i) and new mineralocorticoid receptor antagonists (MRA) can reduce urine protein and reduce renal endpoint events, but both can reduce the urine albumin / creatinine ratio (UACR) by less than 400 mg / g, and the relative risk is only reduced by 16%-39%. The incidence of DKD and the resulting ESKD are still on the rise. Currently, the clinical staging diagnosis of diabetic nephropathy uses the Kidney Disease Improving Global Outcomes (KDIGO) guidelines to recommend combining CKD staging (G1-G5) and albuminuria staging (stage A1: UACR <30 mg / g, stage A2: UACR 30-300 mg / g, stage A3: UACR >300 mg / g) to describe and determine the severity of diabetic nephropathy (it is recommended to test serum creatinine and use the MDRD or CDK-EPI formula to calculate eGFR).

[0003] High sugar is an aggravating factor of the disease and induces premature renal aging. The pathological development of DKD is characterized by overlapping structural and functional changes. Structural changes - mainly GBM thickening, increased membrane matrix, reduced podocyte loss, and Kimmelstiel-Wilson nodules produce functional changes - involving metabolic pathway disorders, hemodynamic changes, inflammatory cascades, growth factor-fibrosis pathways, and genetic susceptibility. Previous studies have confirmed that age is an important cause of the progression of DKD. The prevalence of CKD in diabetic patients aged ≥65 years is as high as 58.7%, and the older the age, the later the stage. Peer scholars have also confirmed that high sugar and premature renal aging are mutually causal and aggravate the progression of DKD. The team previously confirmed that the mechanism of high sugar involvement in premature renal aging and diabetic nephropathy mainly involves: activation of multiple stress resistance responses, abnormal proteins related to nutrient sensing pathways, downregulation of anti-aging related protein factors, and increase in pro-aging factors. In addition, proteinuria is one of the most common clinical symptoms of kidney disease. Causes such as destruction of the glomerular filtration barrier, impaired tubular reabsorption function, abnormal increase and filtration of small molecular weight proteins in plasma, increased renal excretion or tissue damage and release into urine can all cause varying degrees of proteinuria. Proteinuria is not only a pathological product of kidney disease, but also an independent risk factor for the development of kidney disease. There are two mechanisms by which proteinuria damages the kidneys: excessive accumulation of macromolecular substances filtered through the glomerular basement membrane in the mesangial area causes mesangial cell damage, promotes proliferation and increased matrix synthesis, and leads to mesangial sclerosis. Macromolecular proteins enter the renal tubules and compete with small molecular proteins for reabsorption by renal tubular epithelial cells. Therefore, proteinuria has obvious mesangial toxicity. Excessive increase in protein content increases lysosomal activity, causes the brush border of the renal tubules to fall off, and directly damages the integrity of the structure of the renal tubular epithelial cells. Proteinuria is an independent risk factor for tubulointerstitial damage, and its kidney damage is mainly caused by promoting glomerular sclerosis, tubular damage, and infiltration and fibrosis of inflammatory cells in the renal interstitium. Glomerular sclerosis and tubulointerstitial fibrosis (renal fibrosis) are the pathological basis of diabetic nephropathy and are also the common pathways for almost all kidney diseases to progress to end-stage renal failure, but there is currently no effective treatment. In order to slow down and treat the development of kidney disease, early diagnosis, reduction of proteinuria and reversal of urine UACR are particularly important for diabetic nephropathy. Inhibiting premature renal aging not only embodies the idea of ​​traditional Chinese medicine to prevent disease from changing, but can also effectively reduce kidney damage from the source. Not only that, the pathological manifestations of dampness and turbidity in traditional Chinese medicine theory are closely related to the clinical manifestations of lipid metabolism disorders in modern medicine. The development of drugs that effectively reduce proteinuria, improve lipid metabolism, reduce or reverse the progression of urine UACR, and inhibit the mechanism of premature renal aging with high safety is still a dilemma in the current new drug development in the field of diabetic nephropathy, and is also a research goal. Summary of the invention

[0004] The primary purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide a Chinese medicine composition for preventing and treating elevated UACR.

[0005] Another object of the present invention is to provide an application of the above-mentioned traditional Chinese medicine composition for preventing and treating elevated UACR.

[0006] The objective of the present invention is achieved by the following technical scheme: a traditional Chinese medicine composition for preventing and treating elevated UACR, comprising the following components by mass: 40-60 parts of astragalus, 25-35 parts of euryale, 18-22 parts of Rosa laevigata, 4-6 parts of leech, 18-22 parts of Caulis Sinomenii, and 35-55 parts of Herba Cynoglossi; preferably, the composition comprises the following components by mass: 50 parts of astragalus, 30 parts of euryale, 20 parts of Rosa laevigata, 5 parts of leech, 20 parts of Caulis Sinomenii, and 45 parts of Herba Cynoglossi.

[0007] The use of the above-mentioned Chinese medicine composition for preventing and treating elevated UACR in the preparation of a drug for reversing UACR in the early stage of kidney disease or preventing and treating elevated UACR; preferably comprises the following steps: extracting the above-mentioned Chinese medicine composition for preventing and treating elevated UACR, and the obtained extract is a drug for reversing UACR in the early stage of kidney disease or preventing and treating elevated UACR.

[0008] The extract is preferably an aqueous extract.

[0009] The extract is preferably obtained by method I, method II or method III; wherein method I consists of step (1), method II consists of step (1) and step (2), and step III consists of step (1), step (2) and step (3);

[0010] Step (1): decocting the above-mentioned Chinese medicine composition for preventing and treating elevated UACR with water, separating the solid from the liquid, and obtaining a decoction and a medicinal residue; re-decocting the medicinal residue for more than 0 times to obtain a decoction, wherein the re-decocting is for better extracting the effective components of the recipe for tonifying the kidney and eliminating turbidity; combining the decoctions to obtain a liquid extract;

[0011] Step (2): concentrating the decoction obtained in step (1) to obtain a semi-solid extract;

[0012] Step (3): adding ethanol to the extract obtained in step (2), stirring to obtain a drug ethanol solution, wherein the ethanol concentration in the drug ethanol solution is 40-80% by volume; standing, filtering, obtaining a filtrate and a precipitate; washing the precipitate with an ethanol solution with a concentration of 40-80% by volume for more than 0 times, filtering, obtaining a filtrate, wherein the purpose of washing the precipitate is to more fully extract the active ingredients in the precipitate; combining the filtrates; concentrating the filtrates to obtain a semi-solid extract.

[0013] In step (1):

[0014] The amount of water is preferably calculated based on a solid-liquid ratio of the Chinese medicine composition to water of 1:2 to 1:15 (g:mL); more preferably, calculated based on a solid-liquid ratio of the Chinese medicine composition to water of 1:3 to 4 (g:mL).

[0015] The decoction time is preferably 30 to 60 minutes, and more preferably 30 to 40 minutes.

[0016] The amount of water used in the re-decoction is preferably calculated based on a solid-liquid ratio of the Chinese medicine composition to water of 1:2 to 1:15 (g:mL); more preferably, calculated based on a solid-liquid ratio of the Chinese medicine composition to water of 1:3 to 4 (g:mL).

[0017] The re-decoction time is preferably 30 to 60 minutes, and more preferably 30 to 40 minutes.

[0018] The number of re-decoctions is preferably 1 to 3 times, more preferably 1 to 2 times.

[0019] In step (3):

[0020] The ethanol concentration in the drug ethanol solution is preferably 60% by volume.

[0021] The washing is preferably performed using an ethanol solution having a concentration of 60% by volume.

[0022] The number of washings is preferably 3 times.

[0023] The UACR refers to the urine albumin / creatinine ratio. Normal UACR is less than 30 mg / g. Increased urine UACR is an indicator of animal and human function caused by early and middle stages of diabetic nephropathy.

[0024] The kidney disease is preferably diabetic nephropathy.

[0025] The dosage form of the medicine is decoction, pill, powder, paste, pill, granule, oral liquid, capsule, tablet or injection.

[0026] The medicine is a common preparation, a sustained-release preparation, a controlled-release preparation, a targeted preparation or a microparticle drug delivery system.

[0027] Network disease is a unique part of the theoretical system of traditional Chinese medicine. Professor Liu Weijing, a young Qihuang scholar from Dongzhimen Hospital, led a team to inherit and carry forward the theory of "micro-symptoms in kidney network" of Professor Lu Renhe, a master of traditional Chinese medicine, and the theory of "differentiation of mechanism and treatment" of Professor Wang Yaoxian, a Qihuang scholar, and proposed the idea of ​​"three states of kidney network" to diagnose and treat diabetic nephropathy. Kidney network refers to the collaterals of the kidney system. Whether it is the distribution characteristics of glomerular blood vessels or the filtration barrier function of the renal corpuscle, they all conform to the structural characteristics of the collaterals branching out from the meridians, subdividing layer by layer, and gradually becoming narrow and tortuous. Its physiological characteristics (gathering, retention, communication, and transformation) are also consistent with the functional characteristics of the exchange of body fluids and blood and nutrient metabolism, and the pathological characteristics of the kidney network are condensed more comprehensively as easy to be empty and full, easy to stagnate and stagnate, easy to seep and overflow, easy to enter but difficult to exit, and easy to accumulate. Because the kidney collaterals are small, narrow, and blood in the collaterals moves slowly, if the qi and blood in the meridians are slightly deficient, the qi and blood in the collaterals will be obviously insufficient, resulting in "empty collaterals"; and the dampness and fire (heat) of the six evils enter the kidney collaterals with wind as the forerunner, so that the "evil evils cannot be discharged" and hide in the empty collaterals, blocking qi and hindering body fluids, accumulating heat, thus burning body fluids into phlegm, and refining blood into blood stasis. Therefore, diabetic nephropathy belongs to the category of collateral diseases, and the disease is located in the kidney collaterals. "Kidney deficiency and dampness (turbidity) accumulation" is the core pathogenesis of diabetic nephropathy in the early stage, and "evil evil blocking the kidney" is the key link in the late development of diabetic nephropathy.

[0028] Through clinical exploration and practice, the team created a method of tonifying the kidney and eliminating turbidity and optimized a prescription for tonifying the kidney and eliminating turbidity for clinical prevention and treatment of diabetic nephropathy (DN): 50g of Astragalus, 30g of Euryale, 20g of Rosa rugosa, 5g of Hirudo, 20g of Caulis Sinomenii, and 45g of Herba Astragali. In the prescription, Astragalus and Herba Astragali are used as the main drugs. Astragalus is the best among all the Qi-tonifying drugs, which can tonify the Qi of the spleen and kidney and consolidate the essence and micro-substances. "Compendium of Materia Medica" says, "For those who tonify the kidney, Qi is the mother of water." Tonifying Qi can promote blood circulation and promote water excretion. The three complement each other and can effectively treat blood stasis and water retention caused by Qi deficiency. Raw Astragalus has the nature of lifting and lifting, moving but not staying, and can promote evil to the outside, unblock the meridians and relieve numbness. Herba Astragali can clear the heat of the kidney meridians, while promoting water excretion and removing dampness, promoting blood circulation and removing blood stasis, and detoxifying, and can eliminate the pathological products such as phlegm, blood stasis, and toxins that bind the kidney meridians. Gorgon fruit and golden cherry fruit are the assistant drugs. The combination of the two is Shuilu Erxiandan, which has the effects of tonifying the kidney and strengthening the spleen, consolidating the essence and stopping nocturnal emission, removing dampness and stopping leukorrhea. It has a good therapeutic effect on proteinuria caused by spleen deficiency, the inability to transport and distribute the essence of water and grain, and the inability to open and close the kidneys and consolidate and control. Leeches can break blood and dredge the meridians, remove blood stasis and eliminate symptoms; Qingfengteng can dispel wind and dampness, remove blood stasis and dredge the meridians. The two are adjuvant drugs. Insects and vines can play a role in searching for and removing the hidden evil in the meridians, leading the power of other drugs to reach the kidney meridians directly. The whole formula of Astragalus greatly replenishes the Qi of the spleen and kidney, and together with Euryale ferox and Rosa laevigata, it plays the role of consolidating essence and stopping nocturnal emission. The Herba Astragali clears away heat and detoxifies, promotes diuresis and eliminates dampness, and together with Hirudo and Caulis Sinensis, it promotes blood circulation and removes blood stasis, eliminates symptoms and dredges the meridians, replenishes the deficiency of kidney Qi and the deficiency of kidney essence, eliminates dampness, stasis and other pathological products, replenishes the deficiency and eliminates the excess, strengthens the body and eliminates evil, treats both the deficiency and the excess, and takes into account both the symptoms and the root causes. This formula was obtained by the team based on the previous Yiqi Tongluo Xiezhu formula, combined with the characteristics and target efficacy of diabetic nephropathy, and the seaweed turtle shell in the formula was changed to Euryale ferox and Rosa laevigata. The prescription for replenishing qi, unblocking collaterals and eliminating turbidity is mainly for chronic kidney disease. It uses seaweed to dissipate stagnation and promote diuresis, eliminate phlegm, and is good at dissipating phlegm and softening hard masses, which has the function of dissipating stagnation and eliminating symptoms; it uses turtle shell to nourish yin and reduce fever, soften hard masses and dissipate stagnation, and is good at replenishing yin and qi and breaking up stagnation accumulation, which has the function of benefiting the kidney, unblocking collaterals and eliminating accumulation. It focuses on benefiting the kidney, unblocking collaterals, dissipating stagnation and eliminating symptoms; it has a good effect on the decrease in glomerular filtration rate and deterioration of renal function caused by chronic kidney disease, especially the increase in creatinine, and can significantly improve the clinical symptoms of patients with chronic kidney disease. This prescription is mainly for diabetic nephropathy, and its clinical trials are mainly for diabetic patients. In the formula, Gorgon fruit and Cornus officinalis are used to nourish the kidney and strengthen the spleen, while Caulis Scutellariae is used to eliminate dampness and dredge the meridians, search for and eliminate hidden evil, and lead the medicine directly to the site of the disease. The whole formula focuses on nourishing the spleen and kidney, eliminating turbidity and eliminating evil. The therapeutic effect is mainly to reduce and reverse urinary UACR. It has a better effect on reducing trace albumin in urine, and is more targeted at diabetic nephropathy. It has a good effect on increased urinary UACR in the early and middle stages of diabetic nephropathy, lipid metabolism disorders, and different degrees of renal pathological damage; it also has the effect of improving renal function, reducing or reversing urinary UACR, and reducing lipid metabolism disorders.

[0029] Compared with the prior art, the present invention has the following advantages and effects:

[0030] 1. The inventors of the present invention have discovered for the first time that the Bushen Xiezhuo formula has a good therapeutic effect on increased urine UACR, lipid metabolism disorders and different degrees of renal pathological damage in the early and middle stages of diabetic nephropathy; the Bushen Xiezhuo formula also has the effects of improving renal function, reducing or reversing urine UACR, and reducing lipid metabolism disorders.

[0031] 2. The recipe for tonifying the kidney and eliminating turbidity has the effect of nourishing the spleen and kidney and reducing turbidity and toxins. It is widely used in tonifying the kidney and clinical and basic experiments have found that it does not cause liver and kidney damage, so it is relatively safe.

[0032] 3. Compared with its split formula, modified formula and positive drugs, the whole formula of tonifying the kidney and eliminating turbidity has better therapeutic effect and faster recovery in reducing or reversing urinary UACR, and has broad application prospects in the early treatment of diabetic nephropathy. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 This is a graph showing the results of the intervention of the Bushen Xiezhuo recipe, its split recipes, modified recipes and positive drugs in the body weight changes of DN rats.

[0034] Figure 2 This is a graph showing the blood sugar changes in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0035] Figure 3 This is a graph showing the changes in kidney weight / body weight of DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0036] Figure 4 This is a graph showing the changes in 24-hour urinary protein excretion rate of DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0037] Figure 5 This is a graph showing the changes in UACR in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0038] Figure 6 This is a graph showing the changes in blood urea nitrogen in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0039] Figure 7 This is a graph showing the changes in blood creatinine in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0040] Figure 8 This is a graph showing the changes in blood uric acid in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0041] Fig. 9 This is a graph showing the changes in total cholesterol in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0042] Fig.10 This is a graph showing the changes in triglycerides in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0043] Fig.11 This is a graph showing the changes in apolipoprotein in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0044] Fig.12 This is a graph showing the changes in alanine aminotransferase in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0045] Fig.13 This is a graph showing the changes in aspartate aminotransferase in DN rats after intervention with the Bushen Xiezhuo recipe and its split recipes, modified recipes, and positive drugs.

[0046] Fig.14 This is a graph showing the results of the kidney-tonifying and detoxifying prescription and its split prescriptions, modified prescriptions and positive drugs in intervening in the renal pathological changes in DN rats.

[0047] Fig.15 This is a graph showing the results of the Bushen Xiezhuo recipe and its split recipes, modified recipes and positive drugs in intervening in the changes in the mechanism of premature renal aging in DN rats. DETAILED DESCRIPTION

[0048] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0049] Example 1 Clinical study on the treatment of early diabetic nephropathy with the Bushen Xiezhuo recipe

[0050] Subjects: Patients diagnosed with diabetic nephropathy who visited Dongzhimen Hospital of Beijing University of Chinese Medicine from July 2020 to July 2024 were retrospectively analyzed and divided into six groups: simple conventional treatment group, group A, group B, group C, group D, and group E. The conventional treatment group only used conventional Western medicine treatment, and the other groups were treated with Chinese medicine prescriptions on the basis of conventional Western medicine treatment. Group A took Bushen Xiezhuo prescription, group B took Shuilu Erxiandan (Gorgon fruit and golden cherry medicine pair), group C took Bushen Xiezhuo prescription changed to prescription 1 (Ganhuangcao was changed to Tufuling, the dosage remained unchanged), group D took Bushen Xiezhuo prescription changed to prescription 2 (Qingfengteng was changed to Haifengteng, the dosage remained unchanged), group E took Bushen Xiezhuo prescription changed to prescription 3 (the dosage of the main medicine Huangqi and Ganhuangcao was changed, and the total dosage of the whole prescription remained unchanged). In addition, each group was divided into two groups according to UACR 30-300mg / g and 300-600mg / g. A total of 345 patients were treated with Bushen Xiezhuo prescription and related prescriptions for more than 3 months, including 58 patients in the conventional treatment group, 60 patients in the prescription A group, 57 patients in the prescription B group, 55 patients in the prescription C group, 59 patients in the prescription D group, and 56 patients in the prescription E group. There was no statistically significant difference in gender and age among the groups.

[0051] Grouping and drug preparation: A simple conventional treatment group was planned based on the source of prescriptions and drugs (① Reduce urine protein and protect renal function: Irbesartan tablets (Ambovi): 300 mg each time, once a day, orally; ② For blood sugar reduction, the drug should be reasonably selected according to the "Guidelines for the Prevention and Treatment of Diabetes in China". During the trial, the subjects' HbA1C should be maintained at ≤7.5%; ③ Blood pressure should be controlled at ≤130 / 80 mmHg. When blood pressure does not meet the standard, the following drugs should be selected in turn: calcium ion antagonists, β-receptor blockers, and diuretics. Note: ACEI / ARB drugs should no longer be used; ④ For blood lipid reduction, blood lipid reduction treatment should be given according to the 2013 ADA guidelines; ⑤ For other symptomatic treatments, subjects with cardiovascular risks can take aspirin; those with high blood uric acid can take similar drugs. Related drugs), Group A (50g of Astragalus, 30g of Euryale, 20g of Rosa Laevigata, 5g of Leech, 20g of Caulis Sinomenii, 45g of Herba Smilacis Glabrae, combined with conventional western medicine treatment), Group B (30g of Euryale, 20g of Rosa Laevigata, combined with conventional western medicine treatment), Group C (50g of Astragalus, 30g of Euryale, 20g of Rosa Laevigata, 5g of Leech, 20g of Caulis Sinomenii, 45g of Herba Smilacis Glabrae, combined with conventional western medicine treatment), Group D (50g of Astragalus, 30g of Euryale, 20g of Rosa Laevigata, 5g of Leech, 20g of Caulis Sinomenii, 45g of Herba Smilacis Glabrae, combined with conventional western medicine treatment), Group E (30g of Astragalus, 30g of Euryale, 20g of Rosa Laevigata, 5g of Leech, 20g of Caulis Sinomenii, 65g of Herba Smilacis Glabrae, combined with conventional western medicine treatment). Sufficient amount of slices in each group were added with water at a material-liquid ratio of 1:3 (g / mL), decocted twice, each time for 40 min, to obtain the medicinal solution and residue, and the filtrate was combined.

[0052] Western medicine diagnostic criteria:

[0053] Diabetes: The diagnostic criteria of ADA in 2013 are as follows: ① HbA1C ≥ 6.5%; or ② patients with typical symptoms of hyperglycemia or hyperglycemic crisis, with plasma glucose (glucose oxidase method) level > 11.1 mmol / L (200 mg / dl) at any time; or ③ fasting plasma glucose (FBG) level > 7.0 mmol / L (126 mg / dl), where fasting is defined as no caloric intake for at least 8 hours; or ④ in the OGTT test, 2hBG level > 11.1 mmol / L (200 mg / dl); or patients currently taking hypoglycemic drugs.

[0054] Diabetic kidney disease: The 2012 K-DOQI standard of the American Kidney Foundation is used: patients with macroalbuminuria or microalbuminuria with diabetic retinopathy, or type 1 diabetes for more than 10 years with microalbuminuria can mostly be diagnosed with diabetic kidney disease. However, other chronic kidney diseases are considered in the following situations: no diabetic retinopathy; rapid decrease in GFR; rapid increase in proteinuria or nephrotic syndrome; refractory hypertension; urine sediment activity (red blood cells); symptoms or signs of other systemic diseases; GFR decrease of more than 30% within 2 to 3 months after starting ACEI / ARB treatment.

[0055] Proteinuria refers to the excretion of more than normal amounts of protein in the urine. The classification criteria for proteinuria are shown in Table 1, depending on the urine sample.

[0056] Table 1

[0057] Staging Random sample (mg / g) Normoalbuminuria <30 Microalbuminuria 30-300 Macroalbuminuria >300

[0058] Inclusion criteria:

[0059] Meet the diagnosis of diabetic kidney disease (2012 K / DOQI standard): clinical microalbuminuria stage and macroalbuminuria stage, specifically: (1) Meet the diagnosis of diabetic kidney disease (DKD): a) type 2 diabetes; b) combined with diabetic retinopathy; (2) UACR 30mg / g-600mg / g; (3) serum creatinine <265μmol / L (3mg / dl); (4) with hypertension (blood pressure can be controlled to ≤130 / 80mmHg by medication) or without hypertension; (5) HbA1c ≤7.5%; (6) have been taking irbesartan tablets for 2 weeks or more, with a daily dose of 100mg; (7) aged 18 to 70 years, regardless of gender.

[0060] Grouping criteria:

[0061] According to UACR, the patients were divided into a low baseline group (UACR between 30 and 300 mg / g) and a high baseline group (UACR > 300 mg / g).

[0062] Exclusion criteria:

[0063] Patients who meet any of the following conditions are not allowed to participate in the study: patients with primary or secondary kidney diseases other than DKD; patients with urinary system diseases other than DKD (such as urinary tract infection, cystitis, female vaginitis, prostatitis, etc.); patients with other serious diseases (such as malignant tumors, etc.); patients with various infectious diseases in the past month; patients who have received kidney transplants, hematopoietic stem cell transplants or bone marrow transplants; women who are pregnant; and patients with other conditions that the researchers believe are not suitable for inclusion.

[0064] method:

[0065] The study used its own before-and-after control to compare the therapeutic differences between the Bushen Xiezhuo prescription and related proposed prescriptions.

[0066] Main outcome measures:

[0067] Efficacy indicators: UACR, serum creatinine, glomerular filtration rate, cystatin-C;

[0068] Safety indicators: blood potassium, liver function (alanine aminotransferase, aspartate aminotransferase).

[0069] Statistical methods:

[0070] SPSS was used for statistical analysis. The measured data that conformed to the normal distribution were expressed as mean ± standard deviation (x ± s), and the data that did not conform to the normal distribution were expressed as median (interquartile range). For the data that conformed to the normal distribution, paired sample t-test was applied before and after treatment, and one-way analysis of variance was applied for comparison among multiple groups. If the data did not conform to the normal distribution, non-parametric test was applied, and p < 0.05 indicated statistical significance.

[0071] result

[0072] 1) UACR

[0073] The low baseline group was considered to be reversed (effective) when UACR tended to normal; according to previous literature, the drug was effective when the patient's UACR decreased by 30%, so the high baseline group was considered effective when the outcome UACR value decreased by 30% compared with the baseline. According to statistical analysis, the efficacy between the low baseline group and the high baseline group was not completely equal (p < 0.05). After multiple comparisons between the groups, the efficacy of the A prescription group was different from that of other proposed prescriptions (p < 0.05 or p < 0.01). The A prescription had the highest treatment efficacy (reversal rate) for UACR. The specific data are shown in Table 2.

[0074] Table 2 Analysis of the effectiveness of reversing UACR

[0075]

[0076]

[0077] Note: Low baseline group: * represents p < 0.05 compared with group A, ** represents p < 0.01 compared with group A, *** represents p < 0.001 compared with group A; High baseline group: * represents p < 0.05 compared with group A, ** represents p < 0.01 compared with group A.

[0078] 2) Serum creatinine level and glomerular filtration rate level

[0079] Since creatinine (Cr) in the blood is mainly filtered and excreted from the body by the kidneys, the blood creatinine level reflects the function of the kidneys to a certain extent. It is one of the commonly used indicators for diagnosing kidney diseases in clinical practice and is widely used in the diagnosis and monitoring of diseases such as chronic kidney disease, chronic renal failure, acute kidney injury and nephrotic syndrome. The estimated glomerular filtration rate (eGFR) is one of the commonly used indicators for measuring the renal function of the subject in clinical practice. A decrease in this indicator indicates that the patient may suffer from various kidney diseases such as acute kidney injury, chronic renal failure, and diabetic nephropathy. The eGFR is often calculated using the CKD-EPI formula. According to statistical analysis, the efficacy of each prescription group was obtained through a before-and-after comparison t-test, and the efficacy of Group A, Group C, Group D, and Group E was significant (p < 0.0001). Due to uneven variance or data that do not conform to normal distribution, the rank sum test was used to compare the components. There was no significant difference in creatinine and estimated glomerular filtration rate at week 0 (p>0.05), so the baseline of the components can be considered equal. There was a significant difference in creatinine and estimated glomerular filtration rate at week 12 (p<0.0001), so it can be considered that the efficacy of each prescription group is not completely equal. According to multiple comparisons and outcome descriptions, it can be seen that the Bushen Xiezhuo prescription (Group A) is the most effective in reducing patients' blood creatinine and increasing glomerular filtration rate. The effects of A and the proposed prescription on patients' blood creatinine are shown in the following table. When the data is normally distributed, the Expression, the data is in non-normal distribution and quartile expression is used:

[0080] Table 3 Serum creatinine level and glomerular filtration rate level

[0081]

[0082] Note: * represents p<0.05 in self-comparison before and after, ** represents p<0.01 in self-comparison before and after, *** represents p<0.001 in self-comparison before and after, **** represents p<0.0001 in self-comparison before and after; compared with the Bushen Xiezhuo prescription group, ## represents p<0.01, #### represents p<0.0001.

[0083] 3) Cystatin-C

[0084] Cystatin-C (Cys-C) is an endogenous cystine protease inhibitor, and its increase reflects the patient's decreased glomerular filtration function and tubular reabsorption function. Changes in Cys-C are not affected by factors such as inflammation, gender, age, and liver function, and are a common indicator for evaluating renal function. CysC is also a risk factor for the progression of DKD. Relevant studies have shown that Cys-C is positively correlated with the progression of diabetic nephropathy. Elevated Cys-C can be found in patients with early diabetic nephropathy, so Cys-C is also a sensitive indicator for the early diagnosis of diabetic nephropathy. By monitoring the level of Cys-C, we can detect changes in the condition of patients with diabetic nephropathy earlier, and it can also provide a certain basis for predicting the progression of diabetic nephropathy. After statistical analysis, because some data did not conform to the normal distribution, the rank sum analysis was used for component comparison. There was no difference between the low baseline group and the high baseline group at 0 week (p>0.05), and there was a significant difference between the low baseline groups at 12 weeks (p<0.0001), and there was a significant difference between the high baseline groups (p<0.0001). It can be considered that the treatment effects between different groups are not completely equal. Multiple comparisons of components show that the Bushen Xiezhuo prescription has better effect and applicability for lowering patients' serum Cys-C than other split prescription groups. The efficacy of different drug groups was analyzed using the before-and-after t-test or rank sum test. In the low baseline group, there were significant differences between group A (Bushen Xiezhuo group), group D (Bushen Xiezhuo group with Qingfengteng replaced by Haifengteng group), and group E (Bushen Xiezhuo group with changed dosage of the main drug) (p < 0.0001, p < 0.05, p < 0.0001). In the high baseline group, there were significant differences between group A (Bushen Xiezhuo group), group D (Bushen Xiezhuo group with changed Qingfengteng replaced by Haifengteng group), and group E (Bushen Xiezhuo group with changed dosage of the main drug) (p < 0.0001, p < 0.05, p < 0.01). The effects of Bushen Xiezhuo and the proposed prescription on cystatin-C in patients are shown in the following table. When the data is normally distributed, the Expression, non-normal data, expressed according to the quartile method:

[0085] Table 4 Clinical serum cystatin-C status

[0086]

[0087] Note: * represents p<0.05 in self-comparison before and after, ** represents p<0.01 in self-comparison before and after, *** represents p<0.001 in self-comparison before and after, **** represents p<0.0001 in self-comparison before and after; compared with the Bushen Xiezhuo prescription group, ## represents p<0.01, #### represents p<0.0001.

[0088] 4) Safety indicators:

[0089] During the treatment, the vital signs of the five groups of patients were stable. After 12 weeks of treatment, liver function, electrolytes and other indicators were tested to evaluate the effects of the Bushen Xiezhuo prescription and the proposed prescription on the life safety of patients. Compared with before treatment, there were no obvious abnormalities in blood potassium, AST, ALT and other indicators, and no indicators exceeded the normal range, indicating that the Bushen Xiezhuo prescription has good safety, no obvious toxic side effects, and is not prone to adverse reactions (see Tables 5-7 for details).

[0090] 4-1) Blood potassium (mmol / L):

[0091] Table 5 Changes in blood potassium (mmol / L)

[0092]

[0093] 4-2)AST (U / L):

[0094] Table 6 Changes of aspartate aminotransferase (AST) before and after treatment

[0095]

[0096] 4-3) ALT (U / L):

[0097] Table 7 Changes of alanine aminotransferase (ALT) before and after treatment

[0098]

[0099] Example 2 Comparison of the Bushen Xiezhuo Recipe and Tongluo Xiezhuo Recipe in the Treatment of Early Diabetic Nephropathy

[0100] Subjects: Same as Example 1; 60 cases were included in the Bushen Xiezhuo prescription group, and 52 cases were included in the Tongluo Xiezhuo prescription group (also known as Yiqi Tongluo Xiezhuo prescription).

[0101] Grouping and preparation: Bushen Xiezhuo prescription group (50g of Astragalus, 30g of Euryale, 20g of Rosa Laevigata, 5g of Hirudo, 20g of Caulis Sinomenii, 45g of Herba Aspergiae, combined with conventional western medicine treatment), Tongluo Xiezhuo prescription group (40g of Astragalus, 5g of Hirudo, 60g of Herba Aspergiae, 15g of Sargassum, 15g of Carapax Trichosanthis, combined with conventional western medicine treatment). Sufficient pieces of each group were added with water at a material-liquid ratio of 1:3 (g / mL), decocted twice, each time for 40min, to obtain the medicinal solution and residue, and the filtrate was combined.

[0102] Western medicine diagnostic criteria: same as in Example 1.

[0103] Inclusion criteria: Same as Example 1.

[0104] Grouping criteria: According to UACR, patients were divided into a low baseline group (UACR 30-300 mg / g) and a high baseline group (UACR>300 mg / g).

[0105] Exclusion criteria: Same as Example 1.

[0106] method:

[0107] The study used its own before-and-after control to compare the therapeutic differences between the Bushen Xiezhuo prescription and the Tongluo Xiezhuo prescription.

[0108] Main outcome measures

[0109] Efficacy indicators: UACR, serum creatinine, glomerular filtration rate, and cystatin-C.

[0110] Safety indicators: blood potassium, liver function (alanine aminotransferase, aspartate aminotransferase).

[0111] Statistical methods: UACR was analyzed using the chi-square test, and paired sample t-test was used for self-control before and after. P < 0.05 indicated statistical significance.

[0112] result

[0113] 1) UACR

[0114] The low baseline group was considered to be reversed (effective) when UACR tended to normal; according to previous literature, the drug was effective when the patient's UACR decreased by 30%, so the high baseline group was considered effective when the outcome UACR value decreased by 30% compared with the baseline. After statistical test, there was no statistical difference between the two groups (p>0.05), but the effective rate of the Bushen Xiezhuo prescription group was significantly higher than that of the Yiqi Tongluo Xiezhuo prescription group, indicating that the Bushen Xiezhuo prescription was better than the Yiqi Tongluo Xiezhuo prescription in reversing UACR, as shown in Table 8.

[0115] Table 8 Analysis of the effectiveness of Bushen Xiezhuo prescription and Yiqi Tongluo Xiezhuo prescription in reversing UACR

[0116]

[0117] 2) Serum creatinine level and glomerular filtration rate level

[0118] According to statistical analysis, both prescriptions can effectively reduce the patient's blood creatinine level and increase the glomerular filtration rate. The effects on the patient's blood creatinine are shown in the following table

[0119] Table 9 Serum creatinine level and glomerular filtration rate level

[0120]

[0121] Note: **** represents before-after comparison p < 0.0001.

[0122] 3) Cystatin-C

[0123] According to statistical analysis, both prescriptions can reduce the serum Cys-C level of patients. There are significant differences between different groups before and after treatment, but no difference between groups. The effects on cystatin-C of patients are shown in the following table.

[0124] Table 10 Clinical serum cystatin-C status

[0125]

[0126] Note: **** represents before-after comparison p < 0.0001.

[0127] 4) Safety indicators:

[0128] During the treatment, the patient's vital signs were stable. After 12 weeks of treatment, liver function, electrolytes and other indicators were tested to evaluate the effects of the Bushen Xiezhuo prescription and the proposed prescription on the patient's life safety. Compared with before treatment, there were no obvious abnormalities in blood potassium, AST, ALT and other indicators, and no indicators exceeded the normal range, indicating that the Bushen Xiezhuo prescription has good safety, no obvious toxic side effects, and is not prone to adverse reactions.

[0129] 4-1) Blood potassium (mmol / L):

[0130] Table 11 Changes in blood potassium (mmol / L)

[0131]

[0132] 4-2)AST (U / L):

[0133] Table 12 Changes of aspartate aminotransferase before and after treatment

[0134]

[0135] 4-3) ALT (U / L):

[0136] Table 13 Changes of alanine aminotransferase before and after treatment

[0137]

[0138] Example 3: Experimental study on the efficacy of the whole prescription for tonifying the kidney and eliminating turbidity and its aqueous extracts in treating diabetic nephropathy in rats

[0139] 1. Experimental Materials

[0140] (1) Experimental animals

[0141] Male SD (Sprague Dawley) rats, 6-7 weeks old, weighing 200±20g, were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., animal production license number: SCXK (Beijing) 2016-0006. All animals were kept in the SPF-level experimental animal barrier environment of the Key Laboratory of Dongzhimen Hospital of Beijing University of Chinese Medicine, use license number: SYXK (Beijing) 2015-0001, maintained with 12 hours of light and dark alternation, and fed with ordinary feed.

[0142] (2) Drugs and reagents

[0143] The following Chinese medicine prescriptions are all from the herbal medicine store of Dongzhimen Hospital of Beijing University of Chinese Medicine: Group A (50g of Astragalus, 30g of Euryale, 20g of Rosa laevigata, 5g of Leech, 20g of Caulis Sinomenii, 45g of Herba Smilacis Glabrae), Group B (102g of Euryale, 68g of Rosa laevigata), Group C (50g of Astragalus, 30g of Euryale, 20g of Rosa laevigata, 5g of Leech, 20g of Caulis Sinomenii, 45g of Smilax glabra), Group D (50g of Astragalus, 30g of Euryale, 20g of Rosa laevigata, 5g of Leech, 20g of Caulis Sinomenii, 45g of Herba Smilacis Glabrae), Group E (30g of Astragalus, 30g of Euryale, 20g of Rosa laevigata, 5g of Leech, 20g of Caulis Sinomenii, 65g of Herba Smilacis Glabrae).

[0144] 2. Experimental methods

[0145] (1) Preparation of Chinese medicine prescriptions

[0146] 1) Chop the medicinal pieces according to the compatibility ratio of each proposed prescription, add water according to the material-liquid ratio of 1:3 (g / mL), soak for 30 minutes, and boil for 30-40 minutes to obtain the medicinal liquid and residue;

[0147] 2) re-decocting the residue twice according to the step of adding water and decocting in step 1); combining the medicinal liquid obtained in step 1) and the aforementioned re-decocting step;

[0148] 3) The medicinal solution obtained in step 2) is concentrated and decocted, and the required liquid volume is calculated according to the concentration of 1.5 g raw medicinal material / mL, and the experimental medicinal solution is obtained by decocting and volatilizing according to the required liquid volume.

[0149] (2) Establishment of animal model:

[0150] 64 SD rats were selected. After one week of adaptive feeding with ordinary feed (weight 200±20g), healthy and good condition rats with normal blood sugar and negative urine protein qualitative test were selected for the experiment. They were randomly divided into sham operation group (8 rats) and modeling group (56 rats). The sham operation group fasted for 12 hours. After anesthesia, the rats were skinned before surgery with conventional iodine disinfection. The skin and muscle layer were cut longitudinally at 2cm below the left rib and 1cm from the midline of the back. The incision was about 1.5cm, fully exposing the left kidney, peeling the left renal capsule, and ligating the renal pedicle. The abdominal cavity was closed and the incision was sutured layer by layer (one stitch every 0.5cm, and each stitch was tied with a surgical knot 4 times). After the muscle layer was sutured, the sutured site was rinsed with penicillin solution and then the cortex was sutured. After the cortex was sutured, the blood was wiped with iodine cotton balls (to prevent biting), and chlortetracycline ointment was applied to the sutured site. 4 rats were placed in a cage, 2 cages per group, and a total of 8 rats. Lay flat until the rats wake up, do not stack them. After the rats woke up from anesthesia, they were given an intraperitoneal injection of 400,000 units of penicillin (800,000 units dissolved in 2 mL of saline, 1 mL per rat) for 3 consecutive days. The 8 rats in the sham operation group were fed with conventional feed, and the 56 rats in the modeling group were fed with high-sugar and high-fat feed. After 2 weeks of feeding, the rats in the modeling group were fasted but not watered for more than 12 hours. Once a week for 3 consecutive weeks, the rats in the modeling group were given a single intraperitoneal injection of STZ 55 mg / kg (1% STZ was obtained by dissolving STZ in 0.1 mol / L, pH 4.5 citrate buffer before use), while the rats in the sham operation group were given an intraperitoneal injection of the same volume of the above citrate buffer. After the injection, an appropriate amount of sucrose was added to the drinking bottle in each cage. After the injection of STZ, blood was collected from the tip of the tail for 3 consecutive days to measure random blood sugar. If it was higher than 16.7 mmol / L, the model was successfully established. If the blood sugar did not meet the standard, the blood sugar was retested after 1 week. If it still did not meet the standard, STZ 55 mg / kg was injected intraperitoneally again. Insulin injection: For those with random blood sugar > 30mmol / L, give 1IU of glargine insulin subcutaneously once every other day for 2-3 times to keep blood sugar above 16.7mmol.

[0151] (3) Experimental grouping and drug administration

[0152] Eight rats in the sham operation group were selected as controls, and the rats with successful modeling were randomly divided into the model group and the medication group, with 8 rats in each group, specifically divided into DN model group, A group, B group, C group, D group, E group, and valsartan group, a total of 7 groups. The Chinese medicine medication group was gavaged once a day, and the Chinese medicine decoction was gavaged according to the drug dosage conversion of experimental animals (calculated according to the body surface area). The Chinese medicine medication group required 15.3g / kg / d of crude drug per day, that is, 10.2mL / kg / d of water extract. The sham operation group and the model group were gavaged with the same amount of drinking water every day. The valsartan group required 7.2mg / kd / d of crude drug per day, which was consistent with the gavage amount of other groups. The drug was dissolved in drinking water by calculation and then gavaged. The medication was continuously used for 10 weeks, by gavage, once a day.

[0153] (4) Detection method

[0154] 1) General observation

[0155] The general condition of the rats, including their mental state, food intake, water intake, urine output, feces output, body weight, body hair, and activity. After administration, the general symptoms were continuously observed to see if the animals showed any abnormal symptoms.

[0156] 2) Indicator monitoring

[0157] At the end of the 12th week, rats were anesthetized and specimens were collected 1 hour after the first intragastric administration. Urine was collected in a metabolic cage, and paraffin sections of abdominal aorta and kidney specimens of rats were taken. Blood glucose, 24-hour urine protein, urine UACR, aspartate aminotransferase, alanine aminotransferase, blood urea nitrogen, blood creatinine, blood uric acid, liver function, total cholesterol, triglycerides, and apolipoprotein A1 were tested in each group of mice.

[0158] 3) Renal tissue pathological observation

[0159] The left kidney was completely cut off and weighed, the capsule was peeled off and the cortex was separated, and the tissue was quickly placed in a cryotube and put into liquid nitrogen and stored in a -80°C refrigerator. The kidney tissue was fixed in 4% paraformaldehyde, dehydrated in ethanol gradient, embedded in paraffin to make wax blocks, sliced, stained with HE, MASSON, and PAS, observed under an optical microscope, and photographed.

[0160] 4) Immunohistochemistry

[0161] Kidney tissues from each group were sliced, dewaxed, hydrated, antigen repaired, and washed. Appropriate amounts of endogenous peroxidase blockers and goat serum working solutions were added for blocking and incubation. Primary antibodies were diluted with antibody diluent, rabbit anti-p21 antibody (1:1000) and rabbit anti-γH2AX antibody (1:1000). Appropriate amounts of primary antibodies were added to cover the tissues for incubation. Reaction enhancement solution and goat anti-rabbit secondary antibody were added. Freshly prepared DAB working solution was used for color development at room temperature and in the dark. The color development degree was observed under a microscope to control the color development time. The reaction was terminated by washing with pure water. Dehydrated with ethanol solution, transferred into xylene, and sealed with neutral gum. Observed, photographed, and analyzed under an optical microscope.

[0162] (5) Statistical methods

[0163] One-way ANOVA was used for animal experiments. P < 0.05 indicated statistical significance, and P < 0.01 indicated a significant difference.

[0164] 3. Results

[0165] (1) General observation of DN rats

[0166] During the experiment, the rats in the sham operation group moved freely, had normal diet and drinking water, had bright eyes, shiny fur, ruddy and shiny tails, gradually gained weight, had normal urine and feces output, had no symptoms of weight loss and abdominal distension, and were in good mental state. The rats in the model group had significantly increased food and water intake, significantly increased urine output, muscle weight loss, dry and dull hair, listlessness, slow reaction, preference for lying and aversion to movement, and weight loss to varying degrees. Continuous observation after medication found that the rats gained weight and their spirits gradually improved. The changes in body weight, blood sugar, and kidney weight / body weight of rats in each group before and after administration are shown in Tables 14 and Figure 1-Figure 3 The Bushen Xiezhuo recipe has a better effect on increasing the body weight, lowering blood sugar, and reducing the kidney weight / body weight ratio of rats.

[0167] Table 14 Changes in body weight, blood sugar, kidney weight / body weight of DN rats (S±x)

[0168]

[0169] Note: Compared with the model group, # represents p < 0.05; #### represents p < 0.0001; compared with the A group, * represents p < 0.05, **** represents p < 0.0001.

[0170] (2) 24h urine protein quantification (24hUpro (mg / ml)) and urine UACR of DN rats

[0171] Table 15 and Figure 4-Figure 5 The results showed that 24-hour urine protein excretion rate (24h Upro) and urine protein to creatinine ratio (UACR) are important indicators for measuring urine protein excretion. This study analyzed the urine protein excretion of rats in the sham operation group, model group and various drug intervention groups after 12 weeks of intervention and found that rats with diabetic nephropathy had a large amount of proteinuria, but the drug group could reduce proteinuria. The Bushen Xiezhuo recipe has a better effect in reducing urine protein and increasing UACR.

[0172] Table 15 Changes of 24h urine protein and urine UACR in DN rats

[0173]

[0174] Note: Compared with the model group, # represents p < 0.05, #### represents p < 0.0001; compared with the A group, * represents p < 0.05, ** represents p < 0.01, **** represents p < 0.0001.

[0175] (3) Renal function level of DN rats

[0176] Serum creatinine level is the gold standard for diagnosing renal function in diabetic nephropathy and is of great value in assessing the degree of tubulointerstitial damage; while the metabolism of blood urea nitrogen and blood uric acid in the kidney is mainly related to glomerular filtration. The renal function of rats after treatment is shown in Table 16 and Figure 6-Figure 8 As shown, the model group had increased blood urea nitrogen, blood creatinine and blood uric acid levels compared with the sham operation group. The Bushen Xiezhuo recipe can reduce the levels of blood urea nitrogen, blood creatinine and blood uric acid and improve renal function.

[0177] Table 16 Renal function levels in DN rats

[0178]

[0179] Note: Compared with the model group, ## represents p < 0.01, ### represents p < 0.001, and #### represents p < 0.0001; compared with the A group, * represents p < 0.05, ** represents p < 0.01, and **** represents p < 0.0001.

[0180] (4) Lipid metabolism in DN rats

[0181] When patients with kidney disease have dyslipidemia, it may aggravate the patient's kidney damage. Therefore, when kidney disease is combined with dyslipidemia, it should be actively controlled. The blood lipid profile of rats after treatment is shown in Table 17 and Figure 9-11 As shown in the data, the model group had lipid metabolism disorders compared with the sham operation group. The Bushen Xiezhuo prescription and the proposed prescriptions can reduce the levels of total cholesterol and triglycerides. The effect of the Bushen Xiezhuo prescription is more obvious. The Bushen Xiezhuo prescription can also increase the level of apolipoprotein A1 and improve lipid metabolism.

[0182] Table 17 Lipid metabolism in DN rats

[0183]

[0184] Note: Compared with the model group, # represents p < 0.05, #### represents p < 0.0001; compared with the A group, * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.001, **** represents p < 0.0001.

[0185] (5) Safety indicators of DN rats

[0186] As shown in Table 18 and Figure 12-13 As shown in the figure, compared with the sham operation group at the same time point, the ALT and AST levels of the DN model rats at 12 weeks were not significantly different (p>0.05). Compared with the other groups at the same time point, the ALT and AST levels of the rats in the A group were not significantly different (p>0.05). This shows that the Bushen Xiezhuo recipe and the proposed recipes have certain safety.

[0187] Table 18 Liver function levels in DN rats

[0188]

[0189] (6) Renal pathology of DN rats

[0190] The pathological structure of the glomeruli of rats in each group, HE staining can clearly show the nuclear structure, making the nucleus blue, and the basement membrane, collagen fibers and muscle fibers red; the main component of the basement membrane is glycoprotein, PAS staining can fully show the glycoprotein components in the tissue, making the basement membrane, collagen fibers and muscle fibers red. Diabetic kidney damage can accumulate all kidney structures. Masson staining is a classic connective tissue staining method. According to the different permeability of different tissues, different sizes of anions can be selected to stain collagen fibers, elastic fibers and reticular fibers, respectively, to show different colors and different pathological changes, which can determine the glomerular sclerosis and interstitial fibrosis of the kidney. Proteinuria is the main manifestation of glomerular filtration function defect, which is significantly correlated with the structure of the filtration membrane.

[0191] HE and PAS staining results showed that the glomerular structure of the rats in the sham operation group was clear and no obvious abnormalities were observed; the DKD group and each drug group showed varying degrees of glomerular capillary hypertrophy, GBM thickening, mesangial matrix hyperplasia, inflammatory cell infiltration, decreased podocytes, KW nodule formation, etc.; Masson showed that the kidneys of the rats in the sham operation group had no obvious fibrosis pathology, while the DKD group and each drug group showed varying degrees of glomerular sclerosis and tubular damage, and interstitial fibrosis. It can be seen that after the intervention of each drug, the above pathological manifestations were alleviated to varying degrees compared with the DKD group, such as Fig.14 shown.

[0192] like Fig.15 As shown in Table 19, the expression of P21 (cyclin-dependent kinase inhibitor) and phosphorylated histone γH2AX in the kidneys of DN rats increased, indicating that cell senescence occurred in diabetic kidneys. After intervention with drugs such as the Bushen Xiezhuo Recipe, the above pathological manifestations were alleviated to varying degrees. The Bushen Xiezhuo Recipe can regulate cell senescence and delay disease progression by downregulating the expression of P21 and γH2AX.

[0193] Table 19 Expression levels of P21 and γH2AX in DN rats

[0194]

[0195]

[0196] Note: #### indicates p < 0.0001 compared with the model group; **** indicates p < 0.0001 compared with the A group.

[0197] 4. Conclusion

[0198] The experimental results show that the Bushen Xiezhuo prescription has a good UACR reversal rate and reduction rate for diabetic nephropathy stage A2 UACR 30-300mg / g in clinical practice. Long-term follow-up found that the drug can protect renal function, improve glomerular filtration rate, and has good safety. In animal models, the drug can reduce renal pathological damage in diabetic rats, reduce urinary protein excretion and reduce UACR, and improve renal function. There are significant differences in indicators, and long-term follow-up shows that the drug is better than other prescriptions in reducing the disorder of glucose and lipid metabolism in the late stage of the disease. Long-term observations have found that liver function is stable and safe. This provides a basic basis for the Bushen Xiezhuo prescription to treat increased urinary protein excretion, increased UACR, and glucose and lipid metabolism disorders caused by diabetic nephropathy, and is expected to be developed into a new generation of drugs for the treatment of reversal or reduction of increased urinary UACR.

[0199] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A Chinese medicine composition for preventing and treating elevated UACR, characterized in that The invention is composed of the following ingredients by weight: 40-60 parts of astragalus, 25-35 parts of euryale, 18-22 parts of rosa laevigata, 4-6 parts of leech, 18-22 parts of clematis, and 35-55 parts of herba scutellariae.

2. The Chinese medicine composition for preventing and treating elevated UACR according to claim 1, characterized in that The invention is composed of the following ingredients by mass: 50 parts of astragalus, 30 parts of euryale, 20 parts of Rosa laevigata, 5 parts of leech, 20 parts of Caulis Sinomenii, and 45 parts of Herba Lycopodii.

3. Use of the Chinese medicine composition for preventing and treating elevated UACR according to claim 1 or 2 in the preparation of a drug for reversing UACR in the early stage of renal disease or preventing and treating elevated UACR.

4. The use according to claim 3, characterized in that The method comprises the following steps: extracting the Chinese medicine composition for preventing and treating elevated UACR according to claim 1 or 2, and obtaining the extract as a drug for reversing UACR in the early stage of kidney disease or preventing and treating elevated UACR.

5. The use according to claim 4, characterized in that: The extract is a water extract.

6. The use according to claim 4, characterized in that: The extract is obtained by method I, method II or method III; wherein method I consists of step (1), method II consists of step (1) and step (2), and step III consists of step (1), step (2) and step (3); Step (1): decocting the above-mentioned Chinese medicine composition for preventing and treating elevated UACR with water, separating the solid from the liquid, and obtaining a decoction and a medicinal residue; re-decocting the medicinal residue for more than 0 times to obtain a decoction, wherein the re-decocting is for better extracting the effective components of the recipe for tonifying the kidney and eliminating turbidity; combining the decoctions to obtain a liquid extract; Step (2): concentrating the decoction obtained in step (1) to obtain a semi-solid extract; Step (3): adding ethanol to the extract obtained in step (2), stirring to obtain a drug ethanol solution, wherein the ethanol concentration in the drug ethanol solution is 40-80% by volume; standing, filtering, obtaining a filtrate and a precipitate; washing the precipitate with an ethanol solution with a concentration of 40-80% by volume for more than 0 times, filtering, obtaining a filtrate, wherein the purpose of washing the precipitate is to more fully extract the active ingredients in the precipitate; combining the filtrates; concentrating the filtrates to obtain a semi-solid extract.

7. The use according to claim 6, characterized in that: In step (1): The amount of water is calculated based on the solid-liquid ratio of the Chinese medicine composition to water of 1:2 to 1:15 (g:mL); The decoction time is 30 to 60 minutes; The amount of water used in the re-decoction is calculated based on a solid-liquid ratio of the Chinese medicine composition to water of 1:2 to 1:15 (g:mL); The time of re-decoction is 30 to 60 minutes.

8. The use according to claim 6, characterized in that: In step (3): The ethanol concentration in the drug ethanol solution is 60% by volume; The washing is performed using an ethanol solution having a concentration of 60% by volume.

9. The use according to any one of claims 3 to 8, characterized in that: The kidney disease is diabetic nephropathy.

10. The use according to any one of claims 3 to 8, characterized in that: The dosage form of the medicine is decoction, pill, powder, paste, pill, granule, oral liquid, capsule, tablet or injection; The medicine is a common preparation, a sustained-release preparation, a controlled-release preparation, a targeted preparation or a microparticle drug delivery system.

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