A traditional Chinese medicine composition for preventing and treating elevated UACR and its application

Abstract

The present invention belongs to the field of traditional Chinese medicine, and discloses a traditional Chinese medicine composition for preventing and treating elevated UACR and its application. The traditional Chinese medicine composition is composed of the following components in parts by mass: 40-60 parts of Astragalus membranaceus, 25-35 parts of Euryale ferox Salisb., 18-22 parts of Rosa laevigata Michx., 4-6 parts of Hirudo, 18-22 parts of Sinomenium acutum Rehd. et Wils., and 35-55 parts of Penthorum chinense Pursh. The traditional Chinese medicine composition has the effects of invigorating the spleen and kidney, and reducing turbid toxin, has the effect of protecting the liver, and thus has relatively high safety; moreover, it has a better therapeutic effect on reducing or reversing urine UACR and faster recovery, and has broad application prospects in the field of the treatment of early and middle stages of diabetic nephropathy.

Description

Technical Field

[0001] The present invention belongs to the field of traditional Chinese medicine, and particularly 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 mainly manifested by renal microvascular lesions. Approximately 30%-40% of diabetic patients progress to DKD, which is the leading cause of end-stage renal disease (ESRD). Modern medicine uses multiple risk factor interventions, such as blood pressure lowering, blood sugar and lipid regulation, and renin-angiotensin system inhibitors (RASi), etc., but still it is difficult to inhibit the progression of DKD. Evidence-based evidence shows that sodium-glucose cotransporter 2 inhibitors (SGLT2i) and novel mineralocorticoid receptor antagonists (MRA) can reduce urinary protein and reduce renal endpoint events. However, the reduction of urinary albumin / creatinine ratio (UACR) by them is 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. At present, the clinical staging diagnosis of diabetic nephropathy all adopts the Kidney Disease: Improving Global Outcomes (KDIGO) guidelines, which recommend combining the 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 (serum creatinine is recommended for detection, and the MDRD or CDK-EPI formula is used to calculate eGFR).

[0003] High glucose is a factor exacerbating this disease and induces premature kidney aging. The pathological development of DKD is characterized by overlapping structural and functional changes. Structural changes mainly include thickening of the GBM, increased matrix, loss and reduction of podocyte numbers, and formation of Kimmelstiel-Wilson nodules. Functional changes involve disordered metabolic pathways, 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 glucose and premature kidney aging are mutually causal, exacerbating the disease progression of DKD. The team's previous studies have confirmed that high glucose is involved in the comorbidity mechanism of premature kidney aging and diabetic nephropathy, mainly involving activation of multiple stress resistance responses, abnormal proteins related to the nutrient sensing pathway, downregulation of anti-aging related protein factors, and increased pro-aging factors. In addition, proteinuria is one of the most common clinical symptoms of kidney diseases. Glomerular filtration barrier damage, impaired tubular reabsorption function, abnormal increase and filtration of small molecular proteins in plasma, increased renal excretion or tissue damage and release into urine, etc. can all cause varying degrees of proteinuria. Proteinuria is not only a pathological product of kidney diseases but also an independent risk factor promoting the development of kidney diseases. There are two mechanisms by which proteinuria damages the kidneys: Macromolecular substances filtered through the glomerular basement membrane accumulate excessively in the mesangial area, causing mesangial cell damage, promoting proliferation and increased matrix synthesis, leading to mesangial sclerosis. Macromolecular proteins enter the renal tubules and are reabsorbed by renal tubular epithelial cells competitively with small molecular proteins. Therefore, proteinuria has obvious mesangial toxicity. The excessive increase in protein content increases lysosomal activity and causes the shedding of the renal tubular brush border, directly damaging the integrity of the structure of renal tubular epithelial cells. Proteinuria is an independent risk factor for renal tubular interstitial injury, and its resulting renal damage is mainly to promote glomerulosclerosis, renal tubular injury, and infiltration and fibrosis of inflammatory cells in the renal interstitium. Glomerulosclerosis and renal tubular interstitial fibrosis (renal fibrosis) are the pathological bases of diabetic nephropathy and also the common pathway for almost all kidney diseases to progress to end-stage renal failure, but there is currently no effective treatment. To slow down and treat the development of kidney diseases, for diabetic nephropathy, early diagnosis, reducing proteinuria, and reversing urinary UACR are particularly important. Preventing premature kidney aging not only reflects the idea of preventing disease in traditional Chinese medicine but also can effectively reduce kidney damage from the source. Moreover, the pathological manifestation of damp turbidity in traditional Chinese medicine theory is closely related to the clinical manifestations of lipid metabolism disorders in modern medicine. Developing drugs with high safety that can effectively reduce proteinuria, improve lipid metabolism, reduce or reverse the progression of urinary UACR, and prevent premature kidney aging mechanism is still the dilemma and research goal in the field of new drug research and development for diabetic nephropathy currently. 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 dosage of water is preferably calculated according to the material-liquid ratio of traditional Chinese medicine composition to water being 1:2 to 1:15 (g: mL); more preferably calculated according to the material-liquid ratio of traditional Chinese medicine composition to water being 1:3 to 4 (g: mL).

[0015] The decocting time is preferably 30 to 60 min; more preferably 30 to 40 min.

[0016] The dosage of water in the second decoction is preferably calculated according to the material-liquid ratio of traditional Chinese medicine composition to water being 1:2 to 1:15 (g: mL); more preferably calculated according to the material-liquid ratio of traditional Chinese medicine composition to water being 1:3 to 4 (g: mL).

[0017] The time of the second decoction is preferably 30 to 60 min; more preferably 30 to 40 min.

[0018] The number of times of the second decoction 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 carried out using an ethanol solution with a concentration of 60% by volume.

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

[0023] The UACR refers to the urinary albumin / creatinine ratio. In normal people, UACR < 30 mg / g, and the increase in urinary UACR is an animal and human function index caused by the early and middle stages of diabetic nephropathy.

[0024] The nephropathy is preferably diabetic nephropathy.

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

[0026] The drug is a conventional preparation, sustained-release preparation, controlled-release preparation, targeted preparation or particulate drug delivery system.

[0027] Collateral disease theory is a unique part of the traditional Chinese medicine theoretical system. Professor Liu Weijing, a young Qihuang scholar at Dongzhimen Hospital, led the team to inherit and carry forward the theories of "miniature masses in the kidney collaterals" by the national master of traditional Chinese medicine, Professor Lü Renhe, and the theory of "treatment based on syndrome differentiation of pathogenesis" by Qihuang scholar Professor Wang Yaoxian, and put forward the idea of "three states of kidney collaterals" for the diagnosis and treatment of diabetic nephropathy. The kidney collaterals refer to the collateral vessels of the kidney system. Whether it is the distribution characteristics of glomerular blood vessels or the filtration barrier function of renal corpuscles, they all conform to the structural characteristics of collateral vessels branching horizontally from the meridians, being gradually subdivided layer by layer, and gradually becoming narrow and tortuous in the collateral body. Its physiological characteristics (aggregation, retention, circulation, transformation) also conform to the functional characteristics of the exchange of body fluids and blood and nutritional metabolism, and comprehensively summarize the pathological characteristics of the kidney collaterals as being prone to deficiency and excess, stagnation and stasis, leakage and overflow, easy entry and difficult exit, and easy accumulation into masses. Due to the characteristics of the kidney collaterals being small in collateral body, narrow in collateral channels, and slow in collateral blood flow, therefore, as long as there is a slight deficiency of qi and blood in the meridians, the qi and blood in the collaterals will be obviously insufficient, leading to "emptiness of the collateral vessels"; and the pathogenic factors of dampness and fire (heat) among the six exogenous pathogens enter the kidney collaterals led by wind, resulting in "failure to expel the pathogenic factors", hiding and lying latent in the deficient collaterals, blocking qi and obstructing body fluids, accumulating and transforming into heat, thus burning body fluids into phlegm and refining blood into stasis. Therefore, diabetic nephropathy belongs to the category of collateral diseases, with the disease location in the kidney collaterals. "Kidney deficiency with damp (turbidity) accumulation" is the core pathogenesis in the early and middle stages of diabetic nephropathy, and "latent pathogen blocking the kidney" is the key link in the late stage of the 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 first discovered that Bushen Xiezhuo Formula has good curative effects on the elevated urinary UACR, lipid metabolism disorders, and various degrees of renal pathological damage in the early and middle stages of diabetic nephropathy; Bushen Xiezhuo Formula also has the effects of improving renal function, reducing or reversing urinary UACR, and reducing lipid metabolism disorders.

[0031] 2. Bushen Xiezhuo Formula has the effects of strengthening the spleen and kidney and reducing turbidity and toxins. It is widely used in tonifying the kidney, and no liver or kidney damage has been found in clinical and basic experiments, so its safety is relatively high.

[0032] 3. Compared with its disassembled formulas, variant formulas, and positive drugs, the whole Bushen Xiezhuo Formula has better therapeutic effects in reducing or reversing urinary UACR and faster recovery, and has broad application prospects in the treatment field of early and middle-stage diabetic nephropathy. Description of the Drawings

[0033] Figure 1 It is a graph showing the results of the changes in the body weight of DN rats intervened by Bushen Xiezhuo Formula and its disassembled formulas, variant formulas, and positive drugs.

[0034] Figure 2 It is a graph showing the results of the changes in the blood glucose of DN rats intervened by Bushen Xiezhuo Formula and its disassembled formulas, variant formulas, and positive drugs.

[0035] Figure 3 It is a graph showing the results of the changes in the kidney weight / body weight of DN rats intervened by Bushen Xiezhuo Formula and its disassembled formulas, variant formulas, and positive drugs.

[0036] Figure 4 It is a graph showing the results of the changes in the 24-hour urinary protein excretion rate of DN rats intervened by Bushen Xiezhuo Formula and its disassembled formulas, variant formulas, and positive drugs.

[0037] Figure 5 It is a graph showing the results of the changes in the UACR of DN rats intervened by Bushen Xiezhuo Formula and its disassembled formulas, variant formulas, and positive drugs.

[0038] Figure 6 It is a graph showing the results of the changes in the blood urea nitrogen of DN rats intervened by Bushen Xiezhuo Formula and its disassembled formulas, variant formulas, and positive drugs.

[0039] Figure 7 It is a graph showing the results of the changes in the serum creatinine of DN rats intervened by Bushen Xiezhuo Formula and its disassembled formulas, variant formulas, and positive drugs.

[0040] Figure 8 It is a graph showing the results of the changes in the blood uric acid of DN rats intervened by Bushen Xiezhuo Formula and its disassembled formulas, variant formulas, and positive drugs.

[0041] Figure 9 It is a graph showing the results of the changes in the total cholesterol of DN rats intervened by Bushen Xiezhuo Formula and its disassembled formulas, variant formulas, and positive drugs.

[0042] Figure 10 It is a graph showing the changes in triglyceride levels in DN rats intervened by Bushen Xiezhuo Formula, its disassembled formulas, variant formulas, and positive drugs.

[0043] Figure 11 It is a graph showing the changes in apolipoprotein levels in DN rats intervened by Bushen Xiezhuo Formula, its disassembled formulas, variant formulas, and positive drugs.

[0044] Figure 12 It is a graph showing the changes in alanine aminotransferase levels in DN rats intervened by Bushen Xiezhuo Formula, its disassembled formulas, variant formulas, and positive drugs.

[0045] Figure 13 It is a graph showing the changes in aspartate aminotransferase levels in DN rats intervened by Bushen Xiezhuo Formula, its disassembled formulas, variant formulas, and positive drugs.

[0046] Figure 14 It is a graph showing the pathological changes in the kidneys of DN rats intervened by Bushen Xiezhuo Formula, its disassembled formulas, variant formulas, and positive drugs.

[0047] Figure 15 It is a graph showing the changes in the mechanism of premature kidney failure in DN rats intervened by Bushen Xiezhuo Formula, its disassembled formulas, variant formulas, and positive drugs. Specific implementation methods

[0048] The present invention will be further described in detail below in conjunction with the embodiments and the accompanying drawings of the specification, but the implementation manners of the present invention are not limited thereto.

[0049] Example 1 Clinical study on Bushen Xiezhuo Formula in the treatment of early and middle stages of diabetic nephropathy

[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 6 groups: a simple conventional treatment group, Group A, Group B, Group C, Group D, and Group E. The conventional treatment group only received conventional Western medicine treatment, and the other groups received traditional Chinese medicine formulas in addition to conventional Western medicine treatment. Group A took Bushen Xiezhuo Formula, Group B took Shuilu Erxian Dan (the medicinal pair of Euryale ferox and Rosa laevigata), Group C took Variant Formula 1 of Bushen Xiezhuo Formula (replacing Senecio scandens with Smilax glabra, with the dose unchanged), Group D took Variant Formula 2 of Bushen Xiezhuo Formula (replacing Sinomenium acutum with Piper kadsura, with the dose unchanged), Group E took Variant Formula 3 of Bushen Xiezhuo Formula (changing the doses of the monarch drugs Astragalus membranaceus and Senecio scandens, with the total dose of the whole formula unchanged). In addition, each group was further divided into two groups according to UACR of 30 - 300 mg / g and 300 - 600 mg / g. A total of 345 patients were treated with Bushen Xiezhuo Formula and related formulated formulas for more than 3 months, including 58 patients in the conventional treatment group, 60 patients in Group A, 57 patients in Group B, 55 patients in Group C, 59 patients in Group D, and 56 patients in Group E. There were no statistically significant differences in the gender and age of the patients in each group.

[0051] Grouping and pharmaceutical treatment: According to the source of the prescription drugs, a simple conventional treatment group was established (① To reduce urinary protein and protect renal function: Irbesartan tablets (Aprovel): 300 mg each time, once a day, orally; ② To control blood sugar, drugs were rationally selected according to the "China Guidelines for the Prevention and Treatment of Diabetes". During the experiment, the HbA1C of the subjects was maintained at ≤ 7.5%; ③ Blood pressure should be controlled at ≤ 130 / 80 mmHg. When the blood pressure did not reach the standard, calcium antagonists, β-blockers, and diuretics were selected in sequence. Note: ACEI / ARB drugs could no longer be used; ④ To reduce blood lipids, lipid-lowering treatment was given according to the recommended standards of the 2013 ADA guidelines; ⑤ For other symptomatic treatments, subjects at risk of cardiovascular diseases could take aspirin; those with high blood uric acid could take relevant drugs), Group A (Astragalus membranaceus 50 g, Euryale ferox 30 g, Rosa laevigata 20 g, Hirudo 5 g, Sinomenium acutum 20 g, Penthorum chinense 45 g, combined with conventional Western medicine treatment), Group B (Euryale ferox 30 g, Rosa laevigata 20 g, combined with conventional Western medicine treatment), Group C (Astragalus membranaceus 50 g, Euryale ferox 30 g, Rosa laevigata 20 g, Hirudo 5 g, Sinomenium acutum 20 g, Smilax glabra 45 g, combined with conventional Western medicine treatment), Group D (Astragalus membranaceus 50 g, Euryale ferox 30 g, Rosa laevigata 20 g, Hirudo 5 g, Piper kadsura 20 g, Penthorum chinense 45 g, combined with conventional Western medicine treatment), Group E (Astragalus membranaceus 30 g, Euryale ferox 30 g, Rosa laevigata 20 g, Hirudo 5 g, Sinomenium acutum 20 g, Penthorum chinense 65 g, combined with conventional Western medicine treatment). For each group, an appropriate amount of Chinese herbal pieces was added with water at a material-liquid ratio of 1:3 (g / mL), decocted twice, 40 minutes each time, to obtain the medicinal liquid and the residue, and the filtrates were combined to obtain the final product.

[0052] Western medicine diagnostic criteria:

[0053] Diabetes: The diagnostic criteria of the 2013 ADA were adopted: ① HbA1C ≥ 6.5%; or ② For patients with typical symptoms of hyperglycemia or hyperglycemic crisis, the plasma glucose level (glucose oxidase method) at any time > 11.1 mmol / L (200 mg / dL); or ③ The fasting plasma glucose (FBG) level > 7.0 mmol / L (126 mg / dL), and fasting was defined as no calorie intake for at least 8 hours; or ④ During the OGTT test, the 2hBG level > 11.1 mmol / L (200 mg / dL); or those who were using hypoglycemic drugs.

[0054] Diabetic kidney disease: The K-DOQI criteria of the National Kidney Foundation of the United States in 2012 were adopted: Most patients with macroalbuminuria or microalbuminuria combined with diabetic retinopathy, or those with type 1 diabetes for more than 10 years and microalbuminuria can be diagnosed with diabetic kidney disease. However, other chronic kidney diseases are considered when the following situations occur: no diabetic retinopathy; rapid decrease in GFR; sharp increase in proteinuria or nephrotic syndrome; refractory hypertension; active manifestations in urine sediment (red blood cells); symptoms or signs of other systemic diseases; GFR decreases by more than 30% within 2 to 3 months after starting treatment with ACEI / ARB.

[0055] Proteinuria refers to the excretion of more protein in urine than normal. According to different urine samples, the classification criteria for proteinuria are shown in Table 1.

[0056] Table 1

[0057] Staging Random specimen (mg / g) Normal albuminuria <30 Microalbuminuria 30-300 Macroalbuminuria >300

[0058] Inclusion criteria:

[0059] Meet the diagnosis of diabetic kidney disease (2012 K / DOQI criteria): 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 < 265 μmol / L (3mg / dl); (4) With hypertension (blood pressure can be controlled ≤ 130 / 80 mmHg with medication) or without hypertension; (5) HbA1c ≤ 7.5%; (6) Have continuously taken irbesartan tablets for 2 weeks or more, with a daily dosage of 100mg; (7) Aged 18 - 70 years old, regardless of gender.

[0060] Grouping criteria:

[0061] Patients were divided into a low baseline group (UACR 30 - 300mg / g) and a high baseline group (UACR > 300mg / g) according to UACR.

[0062] Exclusion criteria:

[0063] If any of the following conditions are met, the patient cannot participate in the study: Patients with primary or secondary kidney diseases other than DKD; Patients with other 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 within the past 1 month; Patients who have received kidney transplantation, hematopoietic stem cell transplantation or bone marrow transplantation; Female pregnancy; Other situations considered by other researchers as not suitable for inclusion in the group.

[0064] Method:

[0065] A self - controlled before - and - after study was adopted to compare the treatment differences between the Bushen Xiezhuo Formula and related formulated prescriptions.

[0066] Main observation indicators:

[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. Measurement data that conformed to a normal distribution were expressed as the mean ± standard deviation (x ± s), and data that did not conform to a normal distribution were expressed as the median (inter - quartile range); for data that conformed to a normal distribution, paired - sample t - tests were applied before and after treatment, and one - way analysis of variance was applied for between - group comparisons of multiple groups; if the data did not conform to a normal distribution, non - parametric tests were applied, and p < 0.05 indicated statistical significance.

[0071] Results

[0072] 1) UACR

[0073] In the low - baseline group, the normalization of UACR was considered as reversal (effective); according to previous literature, a 30% reduction in UACR in patients by the drug was considered effective. Therefore, in the high - baseline group, a 30% reduction in the UACR value at the end compared to the baseline was considered effective. After statistical analysis, the curative effects among groups in the low - baseline group and the high - baseline group were not all equal (p < 0.05). After multiple between - group comparisons, the curative effect of Formula A was different from that of other formulated prescriptions (p < 0.05 or p < 0.01). Formula A had the highest treatment effective rate (reversal rate) for UACR. The specific data are shown in Table 2.

[0074] Table 2 Analysis of the effective rate of reversing UACR

[0075]

[0076]

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

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

[0079] Since creatinine (Cr) in the blood is mainly excreted through the kidneys, the blood creatinine level reflects the kidney function to a certain extent. It is one of the commonly used clinical indicators for diagnosing kidney diseases 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. Estimated glomerular filtration rate (eGFR) is one of the commonly used clinical indicators to measure the renal function of the examinee. A decrease in this indicator suggests that the patient may suffer from various kidney diseases such as acute kidney injury, chronic renal failure, and diabetic nephropathy. eGFR is often calculated using the CKD-EPI formula. According to statistical analysis, the efficacy of each prescription group was obtained through paired t-tests before and after. The efficacy of the A prescription group, C prescription group, D prescription group, and E prescription group was significant (p < 0.0001). Due to unequal variances or data that did not conform to the normal distribution, the Wilcoxon rank sum test was used for component comparison. There were no significant differences in creatinine and estimated glomerular filtration rate at week 0 (p > 0.05), so it could be considered that the component baselines were equal. At week 12, there were significant differences in creatinine and estimated glomerular filtration rate (p < 0.0001), so it could be considered that the efficacy among the prescription groups was not all equal. According to multiple comparisons and outcome descriptions, it could be seen that the Bushen Xiezhuo Prescription (A prescription group) was the most effective in reducing the patient's blood creatinine and increasing the glomerular filtration rate. The effects of Prescription A and the proposed prescription on the patient's blood creatinine are shown in the following table. When the data was normally distributed, was expressed, and when the data was not normally distributed, quartiles were used for expression:

[0080] Table 3 Blood Creatinine Level and Glomerular Filtration Rate Level

[0081]

[0082] Note: * represents p < 0.05 for paired comparison before and after; ** represents p < 0.01 for paired comparison before and after; *** represents p < 0.001 for paired comparison before and after; **** represents p < 0.0001 for paired comparison before and after; compared with the Bushen Xiezhuo Prescription group, ## represents p < 0.01, and # 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 process, the vital signs of the five groups of patients were stable. After 12 weeks of treatment, liver function, electrolyte and other indicators were detected to evaluate the impact of Bushen Xiezhuo Decoction and the formulated prescription on the patients' life safety. There were no obvious abnormalities in indicators such as blood potassium, AST, and ALT compared with those before treatment, and no indicators exceeded the normal range, indicating that Bushen Xiezhuo Decoction has good safety, no obvious toxic and 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 in aspartate aminotransferase (AST) before and after treatment

[0095]

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

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

[0098]

[0099] Comparison of Bushen Xiezhuo Decoction and Tongluo Xiezhuo Decoction in the treatment of early diabetic nephropathy in Example 2

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

[0101] Grouping and preparation of medicine: Bushen Xiezhuo Decoction group (Astragalus membranaceus 50g, Euryale ferox 30g, Rosa laevigata 20g, Hirudo 5g, Sinomenium acutum 20g, Penthorum chinense 45g, combined with conventional western medicine treatment), Tongluo Xiezhuo Decoction group (Astragalus membranaceus 40g, Hirudo 5g, Penthorum chinense 60g, Sargassum fusiforme 15g, Trionyx sinensis 15g, combined with conventional western medicine treatment). Each group added water to the sufficient amount of cut herbs according to the ratio of material to liquid 1:3 (g / mL), decocted twice, 40 minutes each time, obtained the medicinal liquid and medicinal residues, and the filtrates were combined to obtain the medicine.

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

[0103] Inclusion criteria: The same as in Example 1.

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

[0105] Exclusion criteria: same as in Example 1.

[0106] Method:

[0107] A self - controlled before - and - after study was adopted to compare the therapeutic differences between the Bushen Xiezhuo Prescription and the Tongluo Xiezhuo Prescription.

[0108] Main observation indicators

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

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

[0111] Statistical method: Chi - square test was used for UACR, and paired - sample t - test was used for self - controlled before - and - after comparison. P < 0.05 indicates statistical significance.

[0112] Results

[0113] 1) UACR

[0114] In the low - baseline group, the normalization of UACR was considered as reversal (effective); according to previous literature, a 30% reduction in UACR by the drug was considered effective. Therefore, in the high - baseline group, a 30% reduction in the UACR value at the end compared to the baseline was considered effective. After statistical testing, 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 superior to the Yiqi Tongluo Xiezhuo Prescription in reversing UACR, as shown in Table 8.

[0115] Table 8 Analysis of the effective rate of the Bushen Xiezhuo Prescription and the Yiqi Tongluo Xiezhuo Prescription in reversing UACR

[0116]

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

[0118] According to statistical analysis, both formula groups could effectively reduce the serum creatinine level of patients and increase the glomerular filtration rate. The impact on the serum creatinine of patients is shown in the following table

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

[0120]

[0121] Note: **** represents p < 0.0001 for self - controlled before - and - after comparison.

[0122] 3) Cystatin - C

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

[0124] Table 10 Clinical serum cystatin-C situation

[0125]

[0126] Note: **** represents p < 0.0001 for comparison before and after treatment with oneself

[0127] 4) Safety indicators:

[0128] During the treatment process, the vital signs of patients were stable. After 12 weeks of treatment, liver function, electrolyte and other indicators were detected to evaluate the effects of Bushen Xiezhuo Prescription and the formulated prescription on the life safety of patients. Indicators such as blood potassium, AST, and ALT showed no obvious abnormalities compared with those before treatment, and no indicators exceeded the normal range, indicating that Bushen Xiezhuo Prescription has good safety, no obvious toxic and 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) situation

[0131]

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

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

[0134]

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

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

[0137]

[0138] Example 3 Pharmacodynamic experiments of the whole Bushen Xiezhuo Prescription, its disassembled and modified prescriptions' water extracts in the treatment of diabetic nephropathy rats

[0139] 1. Experimental materials

[0140] (1) Experimental animals

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

[0142] (2) Drugs and reagents

[0143] The traditional Chinese medicines in the following prescriptions were all from the herbal pharmacy of Dongzhimen Hospital, Beijing University of Chinese Medicine: Prescription A group (Astragalus membranaceus 50 g, Euryale ferox 30 g, Rosa laevigata 20 g, Hirudo 5 g, Sinomenium acutum 20 g, Penthorum chinense 45 g), Prescription B group (Euryale ferox 102 g, Rosa laevigata 68 g), Prescription C group (Astragalus membranaceus 50 g, Euryale ferox 30 g, Rosa laevigata 20 g, Hirudo 5 g, Sinomenium acutum 20 g, Smilax glabra 45 g), Prescription D group (Astragalus membranaceus 50 g, Euryale ferox 30 g, Rosa laevigata 20 g, Hirudo 5 g, Piper kadsura 20 g, Penthorum chinense 45 g), Prescription E group (Astragalus membranaceus 30 g, Euryale ferox 30 g, Rosa laevigata 20 g, Hirudo 5 g, Sinomenium acutum 20 g, Penthorum chinense 65 g).

[0144] 2. Experimental methods

[0145] (1) Preparation of traditional Chinese medicine prescriptions

[0146] 1) The cut crude drugs of each formulated prescription were chopped according to the compatibility ratio, added water according to the material - liquid ratio of 1:3 (g / mL), soaked for 30 min, and decocted for 30 - 40 min to obtain the medicinal liquid and the residue.

[0147] 2) The residue was decocted twice again according to the steps of adding water and decocting in step 1); the medicinal liquids obtained in step 1) and the aforementioned repeated decocting steps were combined.

[0148] 3) The medicinal liquid obtained in step 2) was concentrated by decoction. The required liquid volume was calculated according to the concentration of 1.5 g of crude drugs / mL, and the experimental medicinal liquid was obtained by decocting and volatilizing according to the required liquid volume.

[0149] (2) Establishment of animal models:

[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 methods

[0154] 1) General observation

[0155] The general state of rats, including the mental state, food intake, water intake, urine output, feces output, body weight, body hair, and activity. Continuously observe the general symptoms after administration to check if the animals show any abnormal symptoms.

[0156] 2) Index monitoring

[0157] Anesthetize the rats 1 hour after the last gavage at the 12th week and collect specimens. Collect urine in a metabolic cage and take paraffin sections of the abdominal aorta blood and kidney specimens of the rats. Detect blood glucose, 24-hour urinary protein, urinary UACR, aspartate aminotransferase, alanine aminotransferase, blood urea nitrogen, serum creatinine, blood uric acid, liver function, total cholesterol, triglyceride, and apolipoprotein A1 in each group of mice.

[0158] 3) Histopathological observation of the kidney

[0159] Completely cut off the left kidney, weigh it, strip the capsule to separate the cortex, quickly put it into a cryotube and immerse it in liquid nitrogen, and store it in a -80°C refrigerator. Fix the kidney tissue in 4% paraformaldehyde, dehydrate it with gradient ethanol, embed it in paraffin to make a wax block, section it, and perform HE, MASSON, and PAS staining. Observe and take pictures under an optical microscope.

[0160] 4) Immunohistochemistry

[0161] Take sections of the kidney tissue of each group, dewax, hydrate, repair antigens, wash, add an appropriate amount of endogenous peroxidase blocker, incubate with goat serum working solution for blocking, dilute the primary antibody with antibody diluent, rabbit anti-p21 antibody (1:1000), rabbit anti-γH2AX antibody (1:1000), add an appropriate amount of primary antibody to cover the tissue and incubate. Add reaction enhancer, add goat anti-rabbit secondary antibody, freshly prepare DAB working solution and develop color at room temperature in the dark. Observe the color development degree under the microscope to control the color development time, and wash thoroughly with pure water to terminate the reaction. Dehydrate with ethanol solution, transfer to xylene, and seal with neutral gum. Observe, take pictures, and analyze under an optical microscope.

[0162] (5) Statistical methods

[0163] One-way ANOVA analysis is used for animal experiments. P < 0.05 indicates statistical significance, and P < 0.01 indicates significant differences.

[0164] 3. Results

[0165] (1) General observation of DN rats

[0166] During the experiment, the rats in the sham operation group were active, with normal diet and water intake, bright eyes, shiny fur, rosy and bright tails, gradually increasing body weight, normal urine output and fecal output, no symptoms of emaciation or abdominal distension, and good mental state. The rats in the model group had significantly increased food and water intake, significantly increased urine output, muscle wasting, dull and lusterless hair, listless spirit, slow reaction, liked to lie down and disliked moving, and accompanied by varying degrees of weight loss. After continuous observation after medication, it was found that the body weight of the rats increased and their spirits gradually improved. The changes in body weight, blood glucose and kidney weight / body weight of the rats in each group before and after administration are shown in Table 14 and Figure 1 - Figure 3 as follows. Bushen Xiezhuo Prescription has a better effect on increasing the body weight of rats, reducing blood glucose and reducing the ratio of kidney weight to body weight.

[0167] Table 14 Changes in body weight, blood glucose and 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 prescription group, * represents p < 0.05, **** represents p < 0.0001.

[0170] (2) 24-hour urinary protein quantification (24hUpro (mg / ml)) and urinary UACR of DN rats

[0171] Table 15 and Figure 4 - Figure 5 show that the 24-hour urinary protein excretion rate (24h Upro) and urinary protein creatinine ratio (UACR) are important indicators for measuring urinary protein excretion. Through analyzing the urinary protein excretion of the rats in the sham operation group, model group and each drug intervention group after 12 weeks of intervention in this study, it was found that diabetic nephropathy rats had a large amount of proteinuria, but the drug group could reduce proteinuria. Bushen Xiezhuo Prescription has a better effect on reducing urinary protein and increasing UACR.

[0172] Table 15 Changes in 24-hour urinary protein quantification and urinary UACR of DN rats

[0173]

[0174] Note: Compared with the model group, # represents p < 0.05, # represents p < 0.0001; compared with the A prescription 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 the renal function of diabetic nephropathy and is of great value in evaluating the degree of tubulointerstitial damage. Serum urea nitrogen and uric acid metabolism 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 follows. Compared with the sham operation group, the levels of serum urea nitrogen, serum creatinine, and uric acid were increased in the model group. Bushen Xiezhuo Formula could reduce the levels of serum urea nitrogen, serum creatinine, and uric acid and improve renal function.

[0177] Table 16 Renal function levels of 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 formula 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 exacerbate kidney damage in patients. Therefore, when kidney disease is combined with dyslipidemia, it should be actively controlled. The blood lipid conditions of rats after treatment are shown in Table 17 and Figure 9 - Figure 11 as follows. Compared with the sham operation group, there was lipid metabolism disorder in the model group. Bushen Xiezhuo Formula and each proposed formula could reduce the levels of total cholesterol and triglyceride. The effect of Bushen Xiezhuo Formula was more obvious, and Bushen Xiezhuo Formula could 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, and # represents p < 0.0001; compared with the A formula group, * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.001, and **** represents p < 0.0001.

[0185] (5) Safety indicators of DN rats

[0186] As shown in Table 18 and Figure 12 - Figure 13 as follows. Compared with the sham operation group at the same time point, there was no significant difference in the levels of ALT and AST in the DN model group rats at 12 weeks (p > 0.05). Compared with other groups at the same time point, there was no significant difference in the levels of ALT and AST in the A formula group rats (p > 0.05). It indicates that Bushen Xiezhuo Formula and each proposed formula have certain safety.

[0187] Table 18 Liver function levels of DN rats

[0188]

[0189] (6) Pathological conditions of the kidneys of DN rats

[0190] The pathological structures of the glomeruli of rats in each group. HE staining can clearly show the nuclear structure, staining the nucleus blue, and the basement membrane, collagen fibers, and muscle fibers red; the main component of the basement membrane is glycoprotein, and PAS staining can fully show the glycoprotein components in the tissue, staining the basement membrane, collagen fibers, and muscle fibers red. Diabetic kidney damage can affect all structures of the kidney. Masson staining is a classic connective tissue staining method. By selecting anions of different sizes for staining according to the different permeabilities of different tissues, collagen fibers, elastic fibers, and reticular fibers can be shown as different colors, and different pathological changes can occur, which can be used to judge the glomerulosclerosis and interstitial fibrosis of the kidney. Proteinuria is the main manifestation of glomerular filtration function defects and is significantly related to the structure of the filtration membrane.

[0191] The results of HE and PAS staining showed that: the glomerular structure of rats in the sham operation group was clear and no obvious abnormalities were seen; in the DKD group and each drug group, different degrees of glomerular capillary tuft hypertrophy, thickening of the glomerular basement membrane GBM, mesangial matrix hyperplasia, inflammatory cell infiltration, reduction in the number of podocytes, and formation of KW nodules were seen; Masson staining showed that: no obvious fibrotic pathology was seen in the kidneys of rats in the sham operation group, while different degrees of glomerulosclerosis and tubular damage, and interstitial fibrosis lesions were seen in the DKD group and each drug group. 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, as Figure 14 shown.

[0192] As Figure 15 and Table 19 show, the expressions of P21 (cyclin-dependent kinase inhibitor) and phosphorylated histone γH2AX in the kidneys of DN rats increased, suggesting cellular senescence in diabetic kidneys. After the intervention with drugs such as Bushen Xiezhuo Decoction, the above pathological manifestations were all alleviated to varying degrees. Bushen Xiezhuo Decoction can regulate cellular senescence and delay the progression of the disease by downregulating the expressions 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 formula group.

[0197] 4. Conclusion

[0198] As can be seen from the experimental results, Bushen Xiezhuo Formula has a good reversal rate and reduction rate of UACR for stage A2 diabetic nephropathy with UACR of 30 - 300 mg / g clinically. Long-term follow-up found that this medicine can protect renal function, improve glomerular filtration rate, and has good safety. In the animal model, this drug can reduce the pathological damage of the kidneys in diabetic rats, and has significant differences in reducing urinary protein excretion, decreasing UACR, and improving renal function and other indicators. After long-term follow-up, this drug is superior to other prescriptions in reducing the disorders of glucose and lipid metabolism in the later stage of the disease. Long-term observation found that the liver function is stable and it has safety. This provides a basic basis for Bushen Xiezhuo Formula to treat the elevated urinary protein excretion, elevated UACR, and disorders of glucose and lipid metabolism caused by diabetic nephropathy, and is expected to be developed into a new generation of drugs for reversing or reducing the elevated urinary UACR.

[0199] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A traditional Chinese medicine composition for reversing UACR or preventing and treating the increase of UACR in diabetic nephropathy, characterized in that It is made of the following raw material medicine components by mass parts: 40 - 60 parts of Astragalus membranaceus, 25 - 35 parts of Euryale ferox Salisb., 18 - 22 parts of Rosa laevigata Michx., 4 - 6 parts of Hirudo, 18 - 22 parts of Sinomenium acutum Rehd. et Wils., 35 - 55 parts of Penthorum chinense Pursh.

2. The traditional Chinese medicine composition according to claim 1, wherein It is made of the following raw material medicine components by mass parts: 50 parts of Astragalus membranaceus, 30 parts of Euryale ferox Salisb., 20 parts of Rosa laevigata Michx., 5 parts of Hirudo, 20 parts of Sinomenium acutum Rehd. et Wils., 45 parts of Penthorum chinense Pursh.

3. Use of the traditional Chinese medicine composition according to claim 1 or 2 in the preparation of a drug for reversing UACR or preventing and treating the increase of UACR in diabetic nephropathy.

4. The preparation method of the traditional Chinese medicine composition according to claim 1 or 2, characterized in that It includes the following steps: extracting the traditional Chinese medicine composition according to claim 1 or 2, and the obtained extract is a water extract.

5. The preparation method according to claim 4, characterized in that: The said 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): Decoct the above-mentioned traditional Chinese medicine composition for preventing and treating the increase of UACR with water, carry out solid-liquid separation to obtain a decoction and medicinal residues; decoct the medicinal residues more than 0 times to obtain a decoction; combine the decoctions to obtain an extract in liquid state; Step (2): Concentrate the decoction obtained in step (1) to obtain a semi-solid extract. Step (3): Add ethanol to the extract obtained in step (2), stir to obtain a drug ethanol solution, wherein the ethanol concentration in the drug ethanol solution is 40 - 80% by volume percentage; let it stand, filter to obtain a filtrate and a precipitate; wash the precipitate with an ethanol solution with a concentration of 40 - 80% by volume percentage more than 0 times, filter to obtain a filtrate; combine the filtrates; concentrate the filtrates to obtain a semi-solid extract.

6. According to the preparation method described in claim 5, it is characterized in that: In step (1): The dosage of the said water is calculated according to the material-liquid ratio of the traditional Chinese medicine composition and water being 1:2 - 1:15 (g:mL); The decocting time is 30 - 60 min; The dosage of water in the repeated decoction is calculated according to the material-liquid ratio of the traditional Chinese medicine composition and water being 1:2 - 1:15 (g:mL); The time of the repeated decoction is 30 - 60 min.

7. According to the preparation method described in claim 5, it is characterized in that: In step (3): The ethanol concentration in the said drug ethanol solution is 60% by volume percentage; The said washing is carried out using an ethanol solution with a concentration of 60% by volume percentage.

8. The application according to claim 3, wherein: The said drug is a sustained-release preparation, a controlled-release preparation or a targeted preparation.

9. The application according to claim 3, wherein: The dosage form of the said drug is a decoction, a pill, a powder, an ointment, a pellet, a granule, an oral liquid, a capsule or a tablet.

Images