A traditional Chinese medicine composition for treating diabetic nephropathy, its pharmaceutical preparation and application.

Abstract

This invention relates to the field of traditional Chinese medicine, specifically to a traditional Chinese medicine composition for treating diabetic nephropathy, its preparation method, and its application. The traditional Chinese medicine composition of this invention consists of the following raw materials in parts by weight: 10-60 parts of raw Astragalus membranaceus, 5-30 parts of Polygonatum sibiricum, 5-30 parts of Dioscorea opposita, 5-30 parts of Cornus officinalis, 3-15 parts of Coptis chinensis, 5-30 parts of Euonymus alatus, 5-30 parts of Arctium lappa, and 3-15 parts of Hirudo medicinalis. This invention optimizes the selection of raw materials and their compatibility, focusing on strengthening the spleen and replenishing the middle jiao, invigorating the spleen and benefiting the kidneys, strengthening the spleen and consolidating the kidneys, clearing heat and detoxifying, fully utilizing the synergistic therapeutic characteristics of traditional Chinese medicine with multiple components, pathways, and targets, and can significantly improve the clinical healing of patients with diabetic nephropathy. Preliminary small-scale clinical observations have found that the spleen-strengthening, kidney-benefiting, and heat-clearing formula can effectively improve the clinical symptoms and TCM syndromes of patients with diabetic nephropathy, reduce UACR values, and has good potential for widespread application.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine compositions, specifically to a traditional Chinese medicine composition for treating diabetic nephropathy, its pharmaceutical preparation, and its application. Background Technology

[0002] Diabetic nephropathy (DKD) is a chronic kidney disease (CKD) caused by diabetes mellitus (DM). It is one of the major microvascular complications of DM, characterized by varying degrees of proteinuria and progressive renal function decline. International studies have found that 20%-40% of DM patients also have DKD, and approximately 50% of DKD patients progress to end-stage renal disease (ESRD), requiring dialysis or kidney transplantation. Meta-analysis shows that the prevalence of DKD among type 2 DM patients in my country is 21.8%. DKD has become a leading cause of CKD among hospitalized patients in my country and, after chronic nephritis (CGN), a major cause of ESRD, placing a significant economic burden on society and families.

[0003] Proteinuria is the most important clinical manifestation and pathological product of kidney disease (DKD), and a key factor promoting its development. The initial process of proteinuria formation involves glomerular and tubular damage, and is considered a result of damage to the glomerular filtration barrier or tubulointerstitial system. Proteinuria is not only an indicator of kidney damage but also closely related to the progression of DKD. Persistent proteinuria and leakage of plasma proteins accumulated in the glomeruli are independent risk factors for exacerbating glomerulosclerosis. Furthermore, urinary protein itself has a toxic effect that causes tubulointerstitial damage, inducing the production of inflammatory factors by stimulating tubular epithelial cells (PTECs), leading to inflammation and fibrosis. This is one of the main reasons why DKD develops into endocrine renal disease (ESRD). Therefore, clinical intervention targeting proteinuria in DKD can help delay the progression of DKD to ESRD, and has high economic and social value.

[0004] With advancements in medicine and continuous improvements in renal replacement therapies such as dialysis and kidney transplantation, the efficacy of end-stage renal disease (ESRD) has significantly increased. However, how to reduce proteinuria in disseminated kidney disease (DKD), slow its progression, and prevent it from developing into ESRD remains a challenge for the medical community. Early intervention for DKD patients to reduce proteinuria and prevent ESRD has become a crucial issue for DKD researchers. Traditional Chinese medicine (TCM) possesses multi-target and multi-pathway therapeutic characteristics. Using TCM to treat DKD not only offers broad applicability and fewer side effects but also provides irreplaceable advantages over Western medicine in improving clinical symptoms and slowing DKD progression.

[0005] Although there is no specific disease name "diabetic nephropathy" recorded in ancient Chinese medical texts, based on the clinical manifestations of DKD, it can be categorized under the TCM terms "kidney wasting," "wasting kidney," "lower wasting," "edema," and "turbid urine." Traditional Chinese medicine emphasizes "adhering to the pathogenesis," and contemporary physicians generally believe that DKD is caused by deficiency of both the spleen and kidneys, insufficiency of qi and yin, and the presence of blood stasis, dampness, and pathogenic factors. Its pathogenesis can be summarized as deficiency in the root and excess in the branch. The root deficiency, in terms of the internal organs, mainly involves spleen and kidney deficiency; in terms of qi, blood, yin, and yang, it mainly involves qi deficiency, yin deficiency, and yang deficiency. The branch excess includes blood stasis, dampness, phlegm, fluid retention, toxins, heat, and wind.

[0006] The spleen governs transportation and transformation, ascending of clear qi, and overall control. When spleen qi is abundant, its physiological functions of consolidating and retaining water and essence are normal. If spleen qi is deficient, it lacks the ability to ascend and control, leading to the leakage of essential substances and resulting in proteinuria. The kidneys govern the intake of qi, consolidation and retention, water metabolism, and the two lower orifices (urethra and anus). They receive and store the essence of the five viscera and six bowels. If the kidneys fail to receive qi, qi fails to retain essence, and the kidney gate is not secure, essential substances leak from below, also causing proteinuria in diabetic kidney disease (DKD). The spleen and kidneys, as the foundation of acquired and innate constitutions, have a mutually nourishing relationship. The healthy functioning of the spleen depends on the warmth of the kidneys, and the essence of the kidneys relies on the spleen's transportation and transformation of water and grain essence to maintain its abundance. When the qi of the spleen and kidneys is abundant, the body's function of consolidating and retaining water and essence is normal. Therefore, spleen and kidney qi deficiency is key to the development of proteinuria in DKD, and "strengthening the spleen and replenishing qi" has become the main clinical treatment method for DKD proteinuria.

[0007] The medical text *Yifang Kao* states, "Diseases of the lower burner are attributed to damp-heat." The lingering damp-heat, leading to persistent and difficult-to-treat proteinuria, is a major characteristic of DKD. Dampness, being a yin evil, is characterized by its sticky and stubborn nature, easily obstructing the ascending and descending of qi and readily attacking yin areas, damaging the lower burner. Heat, being a yang evil, easily injures fluids and depletes qi. Spleen and kidney qi deficiency weakens their transformative and transporting functions, leading to an imbalance in the body's fluid metabolism. Over time, dampness stagnates and transforms into heat. The damp-heat adheres, and its downward-moving nature scorches the lower burner, disturbing the seminal chamber. The kidneys' inability to retain essence results in leakage of essential substances through urine. "When blood is not flowing smoothly, it becomes water," highlighting the significant role of blood stasis in the progression of DKD. Spleen and kidney qi deficiency weakens the body's ability to circulate blood, leading to blood stasis. Prolonged illness can penetrate the collaterals, further obstructing the ascending and descending of qi, resulting in impaired qi transformation. Furthermore, blood stasis easily generates heat, damaging true yin and exacerbating kidney deficiency. Meta-analysis suggests that tonifying qi, nourishing yin, and activating blood circulation can serve as an effective supplementary and alternative therapy for DKD, particularly in improving DKD ergometer, serum creatinine (Scr), and plasma glucose levels. Given the importance of dampness, heat, and blood stasis in the pathogenesis of DKD, emphasizing the clearing of damp-heat, resolving blood stasis, and activating the collaterals is crucial in the treatment of DKD.

[0008] In the progression of DKD, the underlying deficiency and the excess of pathogenic factors are mutually causal and cyclical. Spleen and kidney qi deficiency, along with damp-heat and blood stasis, contribute to the disease's prolonged and lingering course, leading to frequent complications. Therefore, "strengthening the spleen and kidneys, clearing away pathogenic factors" has become crucial for DKD treatment. Consequently, a more diversified, optimally dosed, and modernly pharmacologically supported traditional Chinese medicine composition is needed to enhance therapeutic efficacy. Summary of the Invention

[0009] The purpose of this invention is to provide a traditional Chinese medicine composition (Jianpi Yishen Qinghua Heluo Fang) for treating diabetic nephropathy, its preparation method, and the application of this traditional Chinese medicine composition or pharmaceutical preparation in the preparation of drugs for treating diabetic nephropathy.

[0010] The "Spleen-Strengthening, Kidney-Nourishing, and Blood-Activating Formula" is based on the pathogenesis of DKD (Diabetic Kidney Disease) characterized by spleen and kidney qi deficiency and damp-heat stagnation. It is adapted from the "Spleen-Strengthening and Blood-Activating Formula," a formula based on the experience of renowned TCM doctor Professor Cai Gan in treating diabetes mellitus. It consists of Astragalus membranaceus, Polygonatum sibiricum, Dioscorea opposita, Cornus officinalis, Coptis chinensis, Euonymus alatus, Arctium lappa, and Hirudo medicinalis. This herbal composition fully utilizes the synergistic effects of multiple components, pathways, and targets of traditional Chinese medicine, focusing on strengthening the spleen and replenishing qi, tonifying the kidneys and consolidating essence, clearing heat and detoxifying, dispelling wind and dampness, and promoting blood circulation and removing blood stasis. It can significantly improve the clinical efficacy of DKD patients and slow the progression of DKD. Preliminary small-scale clinical observations have found that the Spleen-Strengthening, Kidney-Nourishing, and Blood-Activating Formula can effectively improve the clinical symptoms and TCM syndromes of DKD patients and reduce the UACR (Ultra-Adverse Reproductive Rate) value.

[0011] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating DKD, the composition comprising the following raw materials in parts by weight:

[0012] Raw Astragalus membranaceus 10-60 parts by weight, Polygonatum sibiricum 5-30 parts by weight, Dioscorea opposita 5-30 parts by weight, Cornus officinalis 5-30 parts by weight, Coptis chinensis 3-15 parts by weight, Euonymus alatus 5-30 parts by weight, Arctium lappa 5-30 parts by weight, Hirudo medicinalis 3-15 parts by weight.

[0013] Furthermore, the preferred traditional Chinese medicine composition of the present invention is composed of the following raw materials in parts by weight: 30 parts of raw Astragalus membranaceus, 15 parts of Polygonatum sibiricum, 15 parts of Dioscorea opposita, 15 parts of Cornus officinalis, 6 parts of Coptis chinensis, 15 parts of Euonymus alatus, 15 parts of Arctium lappa, and 6 parts of Hirudo medicinalis.

[0014] This traditional Chinese medicine composition is based on the three principles of tonification, clearing, and unblocking. Astragalus membranaceus (Huangqi) strengthens the spleen and replenishes the middle jiao, raising yang and lifting prolapse; Polygonatum sibiricum (Huangjing) strengthens the spleen and benefits the kidneys, replenishing qi and nourishing yin; Dioscorea opposita (Shanyao) strengthens the spleen and consolidates the kidneys, replenishing deficiency and benefiting essence; Cornus officinalis (Shanzhuyu) tonifies the kidneys, astringes essence, and consolidates the body; Coptis chinensis (Huanglian) clears heat, detoxifies, and strengthens yin; Euonymus alatus (Guijiyu) invigorates blood, removes blood stasis, and unblocks the collaterals; Arctium lappa (Niubangzi) dispels wind, detoxifies, and eliminates dampness and turbidity; and Hirudo medicinalis (Shuizhi) breaks up blood stasis, removes blood stasis, and unblocks the kidney collaterals. The entire formula uses Astragalus membranaceus as the chief ingredient, Polygonatum sibiricum (Huangjing), Dioscorea opposita (Shanyao), and Cornus officinalis (Shanzhuyu) as assistant ingredients, Coptis chinensis (Huanglian) and Euonymus alatus (Guijiyu) as adjuvant ingredients, and Arctium lappa (Niubangzi) and Hirudo medicinalis (Shuizhi) as guiding ingredients, working together to strengthen the spleen, benefit the kidneys, clear away stagnation, and harmonize the collaterals. See the formula diagram for details. Figure 1 .

[0015] Astragalus membranaceus (Huangqi) is sweet and slightly warm in nature. It ranks first among frequently used herbs in traditional Chinese medicine formulas for treating kidney disease (DKD) and has a clear therapeutic effect on DKD. Modern research has found that astragalus polysaccharides and total astragalus saponins can inhibit excessive activation of the renal artery system (RAS), reduce intraglomerular pressure, and decrease the production of urinary mAlb.

[0016] Polygonatum has a sweet taste and neutral properties, and it can invigorate the spleen and replenish qi, as well as nourish the kidneys and replenish essence. Polygonatum polysaccharides, as the main active ingredient of Polygonatum, have been found to have hypoglycemic and antioxidant effects. Its protective mechanism on the kidneys of diabetic rats may be related to its ability to control blood sugar and its anti-fibrotic factors.

[0017] Yam is sweet and neutral in nature, and has the effects of strengthening the spleen and kidneys, nourishing essence and replenishing deficiency. It is one of the most commonly used Chinese medicines for treating diabetic kidney disease (DKD). Mr. Shi Jinmo believes that when Astragalus membranaceus and yam are used together to treat DKD, they can simultaneously regulate and replenish spleen yang and spleen yin, and together achieve the effects of strengthening the spleen and promoting digestion, and astringing essence and stopping leakage.

[0018] Cornus officinalis has a sour and astringent taste and is slightly warm in nature. It is beneficial to the liver and kidneys, astringes essence and qi, and stops collapse. Numerous in vitro and in vivo experimental studies have found that Cornus officinalis not only has a variety of pharmacological effects such as lowering blood sugar, anti-oxidation, and inhibiting inflammatory response, but can also improve renal fibrosis in DKD rats, reduce inflammatory cell infiltration, and alleviate pathological changes in the kidneys.

[0019] Coptis chinensis, bitter and cold in nature, has the effects of clearing heat and drying dampness, purging fire and detoxifying. Network pharmacology studies have found that Coptis chinensis can regulate the inflammation of DKD renal tissue by affecting the AGEs-RAGE signaling pathway. Meta-analysis has found that berberine has multiple effects such as improving insulin resistance, lowering blood sugar, regulating lipids, lowering blood pressure, reducing urinary albumin, anti-inflammation, anti-oxidative stress, and antibacterial properties. In addition, studies have found that berberine has a protective effect against high glucose-induced podocyte damage, can improve podocyte survival rate and reduce podocyte migration rate under high glucose conditions, maintain glomerular filtration barrier function, and can reduce podocyte damage by activating autophagy through AMPK and its downstream signaling pathways.

[0020] Euonymus alatus has a bitter and pungent taste and is cold in nature. It is good at purging stagnant blood and removing blood stasis, and can also promote diuresis, dispel wind, detoxify, and clear heat. Modern research has found that Euonymus alatus has pharmacological effects such as lowering blood sugar, regulating blood lipids, inhibiting allergic reactions, and improving hypoxia tolerance. Network pharmacology and molecular docking studies have found that Euonymus alatus can participate in the regulation of multiple signaling pathways, playing a role in inhibiting inflammatory responses, anti-oxidative stress, anti-hypoxia, anti-apoptosis, and improving renal hemodynamics to delay the progression of DKD.

[0021] Burdock seed has a bitter and cold nature, and possesses the effects of dispelling wind and clearing heat, detoxifying and promoting diuresis. Network pharmacology and molecular docking studies have found that burdock seed can exert anti-inflammatory and oxidative stress-reducing effects through AGEs-RAGE and other signaling pathways to treat DKD. Modern pharmacological studies have confirmed that burdock seed and its extracts have unique advantages in the treatment of DKD. Through single-drug or compound burdock seed preparations, proteinuria in DKD can be effectively reduced, edema can be alleviated, and there are also effects such as lowering blood sugar, lowering blood pressure, regulating lipid metabolism, improving microcirculation, and regulating immunity.

[0022] Leeches are bitter and salty in taste and neutral in nature, possessing the effects of breaking up blood stasis, clearing the meridians, and eliminating masses. Zhang Xichun stated that "its salty taste specifically enters the blood, breaking up blood stasis without harming new blood, and causing no damage to the qi level." Multiple clinical and experimental studies have shown that leeches and their extracts can play a preventive and therapeutic role in DKD through various means, including inhibiting inflammatory responses and oxidative stress, protecting podocytes, improving vascular endothelial function and hemorheology, lowering lipids, anti-fibrinolysis, anti-apoptosis, and regulating metabolic disorders.

[0023] In a second aspect, the present invention provides a pharmaceutical preparation using the above-mentioned traditional Chinese medicine composition for treating DKD as an active ingredient. This pharmaceutical preparation is prepared using conventional preparation methods in the art into commonly used dosage forms in pharmaceuticals, such as decoctions, granules, tablets, capsules, oral liquids, mixtures, or syrups.

[0024] Furthermore, the pharmaceutical formulation of the present invention can be prepared by the following method:

[0025] (A) Preparation of the spleen-tonifying, kidney-tonifying, clearing and purifying decoction using the traditional decoction method: Weigh the Chinese medicinal herbs according to the formula ratio, add drinking water to the water level 2 cm above the herbs, soak for 30 minutes, heat over high heat until the liquid boils, then reduce to low heat and continue to simmer for 30 minutes. Pour out the liquid, add water to the dregs again, decoct again and pour out the liquid (decoction method is the same as before), combine the two liquids, filter and further concentrate to obtain the water extract of the Chinese medicine composition;

[0026] (B) Prepared by adding a pharmaceutically acceptable carrier to the aqueous extract using conventional methods in the art.

[0027] A third aspect of the present invention is to provide the use of the above-mentioned traditional Chinese medicine composition and pharmaceutical preparation in the preparation of a drug for treating DKD.

[0028] The dosage of this invention is a therapeutically effective dose, for example, about 2 g / kg body weight to about 5.5 g / kg body weight per day, with a preferred dose of about 3 g / kg body weight to about 4 g / kg body weight per day. The general duration of treatment is 6-24 months, preferably 6-18 months, and even more preferably 12-18 months. Of course, the duration of treatment must also consider the route of administration and the patient's health condition, which are all within the scope of a skilled physician's expertise.

[0029] Furthermore, the traditional Chinese medicine composition of the present invention can be used as the sole active ingredient for the treatment of DKD, or it can be used in conjunction with other therapeutic agents (such as empagliflozin).

[0030] The role and effect of invention

[0031] The spleen-tonifying, kidney-nourishing, and blood-activating formula of this invention is safe and effective in treating patients with spleen and kidney qi deficiency and damp-heat stagnation type DKD (G1-G3a stage). It can reduce proteinuria, improve the imbalance of glucose and lipid metabolism and inflammation, reduce risk factors for renal vascular disease, protect kidney function, and alleviate symptoms such as fatigue, shortness of breath, frequent urination, soreness of the waist and knees, edema, heaviness of limbs, dull complexion, and rough skin, thereby improving their quality of life. Attached Figure Description

[0032] Figure 1 This invention relates to a formula for strengthening the spleen, benefiting the kidneys, clearing heat, and resolving phlegm. Detailed Implementation

[0033] The present invention will now be described in detail with reference to the embodiments, but the implementation of the present invention is not limited thereto.

[0034] Unless otherwise specified, experimental methods in the following examples are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise noted, the medicinal materials used in the embodiments of this invention can be obtained from Chinese medicinal herb sales companies. Unless otherwise noted, all obtained medicinal materials are processed Chinese medicinal herbs. These processed herbs can be obtained from sales companies or can be obtained through processing.

[0035] This invention provides a traditional Chinese medicine composition for treating diabetic nephropathy, which is made from the following raw materials: 10-60 parts by weight of raw Astragalus membranaceus, 5-30 parts by weight of Polygonatum sibiricum, 5-30 parts by weight of Dioscorea opposita, 5-30 parts by weight of Cornus officinalis, 3-15 parts by weight of Coptis chinensis, 5-30 parts by weight of Euonymus alatus, 5-30 parts by weight of Arctium lappa, and 3-15 parts by weight of Hirudo medicinalis.

[0036] Preferred ingredients: 20-50 parts by weight of raw Astragalus membranaceus, 5-25 parts by weight of Polygonatum sibiricum, 5-25 parts by weight of Dioscorea opposita, 5-25 parts by weight of Cornus officinalis, 3-12 parts by weight of Coptis chinensis, 5-25 parts by weight of Euonymus alatus, 5-25 parts by weight of Arctium lappa, and 3-12 parts by weight of Hirudo medicinalis.

[0037] Further optimized ingredients include: 30-40 parts by weight of raw Astragalus membranaceus, 10-20 parts by weight of Polygonatum sibiricum, 10-20 parts by weight of Dioscorea opposita, 10-20 parts by weight of Cornus officinalis, 3-9 parts by weight of Coptis chinensis, 10-20 parts by weight of Euonymus alatus, 10-20 parts by weight of Arctium lappa, and 3-9 parts by weight of Hirudo medicinalis.

[0038] The optimal combination is: 30 parts by weight of raw Astragalus membranaceus, 15 parts by weight of Polygonatum sibiricum, 15 parts by weight of Dioscorea opposita, 15 parts by weight of Cornus officinalis, 6 parts by weight of Coptis chinensis, 15 parts by weight of Euonymus alatus, 15 parts by weight of Arctium lappa, and 6 parts by weight of Hirudo medicinalis.

[0039] The preparation method of this traditional Chinese medicine composition for treating diabetic nephropathy includes:

[0040] (A) The traditional decoction method was used to prepare the decoction of the spleen-tonifying, kidney-nourishing, clearing and purifying formula: the Chinese medicinal herbs were weighed according to the formula ratio, drinking water was added to the water level 2 cm above the herbs, soaked for 30 minutes, heated over high heat until the liquid boiled, then reduced to low heat and continued to boil for 30 minutes. The liquid was poured out, the dregs were added to water again, and the same method was used to decoct the dregs. The liquid was then poured out, the two liquids were combined, filtered and further concentrated to obtain the water extract of the Chinese medicinal composition.

[0041] (B) A pharmaceutically acceptable carrier is added to the aqueous extract, and tablets, granules, oral formulations, etc., are prepared using conventional methods in the art, as follows:

[0042] Example 1: Preparation of decoction of traditional Chinese medicine composition

[0043] Weigh out 30 parts by weight of raw Astragalus membranaceus, 15 parts by weight of Polygonatum sibiricum, 15 parts by weight of Dioscorea opposita, 15 parts by weight of Cornus officinalis, 6 parts by weight of Coptis chinensis, 15 parts by weight of Euonymus alatus, 15 parts by weight of Arctium lappa, and 6 parts by weight of Hirudo medicinalis.

[0044] Weigh each of the above-mentioned Chinese herbs according to the formula ratio, rinse them with water, soak them in a container for more than 30 minutes, heat them over high heat until the liquid boils, then reduce the heat to low and continue to simmer for 30 minutes. Pour out the liquid, add water to the dregs again, decoct them in the same way, and pour out the liquid again. Combine the two liquids, filter them, and further concentrate them to obtain the decoction of the Chinese herbal composition.

[0045] Example 2: Preparation of granules from a traditional Chinese medicine composition

[0046] Weigh out 30 parts by weight of raw Astragalus membranaceus, 15 parts by weight of Polygonatum sibiricum, 15 parts by weight of Dioscorea opposita, 15 parts by weight of Cornus officinalis, 6 parts by weight of Coptis chinensis, 15 parts by weight of Euonymus alatus, 15 parts by weight of Arctium lappa, and 6 parts by weight of Hirudo medicinalis.

[0047] Weigh the above-mentioned Chinese herbs according to the formula ratio, rinse with water, and soak in a container for at least 30 minutes. Heat over high heat until the liquid boils, then reduce to low heat and continue simmering for 30 minutes. Pour out the liquid, add water to the dregs again, and decoct using the same method. Pour out the liquid again, combine the two decoctions, filter, and further concentrate. Filter for later use. Finally, take the extract and dextrin according to the ratio and draw them into the fluidized bed granulator container to complete the mixing and granulation. Dry at 50℃ for 2 hours. Sieve the dried granules through a 12-14 mesh sieve, and package them to obtain the final product.

[0048] Example 3: Preparation of tablets from traditional Chinese medicine composition

[0049] Weigh out 30 parts by weight of raw Astragalus membranaceus, 15 parts by weight of Polygonatum sibiricum, 15 parts by weight of Dioscorea opposita, 15 parts by weight of Cornus officinalis, 6 parts by weight of Coptis chinensis, 15 parts by weight of Euonymus alatus, 15 parts by weight of Arctium lappa, and 6 parts by weight of Hirudo medicinalis.

[0050] Weigh the above-mentioned Chinese herbs according to the formula ratio, rinse with water, and soak in a container for at least 30 minutes. Heat over high heat until the liquid boils, then reduce to low heat and continue simmering for 30 minutes. Pour out the liquid, add water to the dregs again, and decoct using the same method. Pour out the liquid again, combine the two decoctions, filter, and further concentrate. Filter for later use. Finally, take the extract and dextrin according to the ratio and draw them into the fluidized bed granulator container to complete the mixing and granulation. Dry at 50℃ for 2 hours. Sieve the dried granules through a 12-14 mesh sieve, add magnesium stearate, mix well, and compress into tablets to obtain the final product.

[0051] Example 4: Clinical and Traditional Chinese Medicine Efficacy of Jianpi Yishen Qinghua Heluo Formula in Treating DKD

[0052] 1. Clinical Data

[0053] 1.1 Source of cases

[0054] This study included patients with DKD (G1-G3a stage) diagnosed with spleen and kidney qi deficiency and damp-heat stagnation syndrome. The included cases were patients treated in the outpatient and inpatient departments of Nephrology, Traditional Medicine and Endocrinology at Shuguang Hospital affiliated with Shanghai University of Traditional Chinese Medicine from March to October 2023, and were selected according to the inclusion and exclusion criteria.

[0055] 1.2 Diagnostic criteria

[0056] 1.2.1 Western medicine diagnostic and staging criteria for DKD: formulated according to the "Chinese Guidelines for Clinical Diagnosis and Treatment of Diabetic Kidney Disease" and the "Chinese Guidelines for Prevention and Treatment of Diabetic Kidney Disease (2021 Edition)".

[0057] 1.2.2 Diagnostic criteria for DKD in Traditional Chinese Medicine: The diagnostic criteria for DKD with spleen and kidney qi deficiency and damp-heat stagnation syndrome were formulated with reference to the "Guiding Principles for Clinical Research of New Traditional Chinese Medicine (Trial)" and the "Standards for Diagnosis, Differentiation of Syndromes and Evaluation of Efficacy of Diabetic Nephropathy (Trial Scheme)".

[0058] 1.3 Inclusion Criteria: Patients aged 18-80 years, regardless of gender; meeting the diagnostic criteria for DKD stages G1-G3a and A2-A3; stable, non-dialysis patients; diagnosed with spleen and kidney qi deficiency and damp-heat stagnation syndrome according to Traditional Chinese Medicine (TCM) principles; blood pressure ≤130 / 80 mmHg, diastolic blood pressure ≥70 mmHg; no mental illness, clear consciousness, and capable of independent judgment; no participation in other drug clinical trials within 3 months prior to enrollment; understanding of the entire trial process, willingness to accept treatment, and signing of an informed consent form. (Note: All of the above criteria must be met for enrollment.)

[0059] 1.4 Exclusion Criteria: Patients with proteinuria caused by DM combined with NDKD (primary kidney disease or other secondary kidney diseases besides DKD); patients with T1DM, gestational DM-related kidney disease, or secondary DM-related kidney disease; patients who have experienced acute complications of DM within the past month, such as DKA, or have undergone surgery, trauma, or other stressful events; patients with serious primary diseases or complications of the cardiovascular, respiratory, hepatic, or hematopoietic systems; patients with malignant tumors, tuberculosis, or other wasting diseases or other acute or chronic infectious diseases; patients with urinary tract infections; patients with massive proteinuria (24hUP > 3.5g); patients with allergies or intolerances to any component of the drugs used in this study; patients who have used nephrotoxic drugs such as nonsteroidal anti-inflammatory drugs or iodine contrast agents within the past 3 months; patients who have newly started using ARBs or ACEIs within the past 3 months; pregnant, breastfeeding, or planning to become pregnant soon. Note: Patients meeting any of the above criteria should be excluded.

[0060] 1.5 Exclusion criteria: Incomplete data collection; participation in other clinical trials during treatment.

[0061] 1.6 Criteria for Trial Termination: Severe adverse reactions or allergic reactions to the test drugs during the trial; failure to follow the trial protocol or adherence to medical advice; eGFR < 30 ml / min / 1.73 mcg. 2 ; Scr doubles or dialysis treatment begins; patient requests withdrawal from trial; serious cardiovascular or cerebrovascular disease occurs during trial.

[0062] 2. Research Methods

[0063] 2.1 Sample size calculation, randomization and grouping methods

[0064] Using UACR as the primary endpoint, a superiority test was employed to estimate the sample size. Based on preliminary clinical observation and experimental data, the mean decrease in UACR after treatment was used as the basis for sample size estimation. PASS 15.0 software was used to calculate the sample size, considering an estimated 10% dropout rate, resulting in a total sample size of 104 cases across both groups. A random number table was generated using SPSS 28.0 statistical software. Patients were randomly assigned to the control and treatment groups in a 1:1 ratio according to the table. A treatment allocation table with serial numbers 1-104 was created. Patients meeting the inclusion criteria were assigned to the control and treatment groups according to their consultation order and serial number, with 52 patients in each group.

[0065] 2.2 Treatment methods

[0066] 2.2.1 Basic Treatment Plan

[0067] 1. General treatment: 1) Dietary control: Control calorie intake, with daily protein intake of about 0.6-0.8g / kg / day and sodium intake of 1.5-2.0g / day (equivalent to sodium chloride 3.75-5.00g / day); 2) Lifestyle adjustment: Moderate exercise, weight control, quitting smoking or reducing the amount of smoking.

[0068] 2. Blood glucose control: Maintain the original blood glucose control regimen. For the patients in this study, the blood glucose control target is HbA1c ≤ 7%. For elderly patients, the HbA1c control target can be appropriately relaxed to 8.5%. For those at risk of hypoglycemia, the HbA1c control target can be further relaxed, but should not exceed 9%. For those whose blood glucose control does not meet the target, insulin therapy can be added. Monitor blood glucose during treatment and be alert to the occurrence of hypoglycemia. If hypoglycemia occurs, adjust the blood glucose control regimen in a timely manner.

[0069] 3. Control blood pressure: Maintain the original antihypertensive regimen (losartan potassium tablets are the first choice for antihypertensive drugs). ARBs or ACEIs should be taken orally for more than 3 months before enrollment. The target blood pressure should be controlled below 130 / 80 mmHg, and the diastolic blood pressure should not be lower than 70 mmHg. For those whose blood pressure does not reach the target after taking antihypertensive drugs, antihypertensive drugs that do not affect the urinary protein excretion rate can be added.

[0070] 4. Control blood lipids: Maintain the original lipid-lowering regimen, with the target LDL value controlled below 2.6 mmol / L. Statins are the first choice for lipid-lowering drugs. Patients in G3 stage should reduce the dosage of pravastatin.

[0071] 5. Correct electrolyte imbalance and regulate water metabolism: For patients with electrolyte imbalance or significant edema, timely correction of electrolyte imbalance and diuretic treatment should be given.

[0072] 6. Other treatments: If other symptoms occur during the trial, provide timely symptomatic treatment and keep a record of the trial.

[0073] 2.2.2 Control group treatment regimen: Basic treatment plus empagliflozin (manufacturer: Boehringer IngelheimPharma GmbH & CO.KG, approval number: HJ20201008), 10mg / tablet, once daily, one tablet each time, orally.

[0074] 2.2.3 Treatment regimen for the treatment group: The treatment regimen for the control group was supplemented with a spleen-tonifying, kidney-nourishing, and blood-activating formula (30g of raw Astragalus membranaceus, 15g of Polygonatum sibiricum, 15g of Dioscorea opposita, 15g of Cornus officinalis, 6g of Coptis chinensis, 15g of Euonymus alatus, 15g of Arctium lappa, and 6g of Hirudo medicinalis), one dose per day, decocted twice in water, the decoctions were combined and taken warm in the morning and evening.

[0075] 2.2.4 Treatment course: Both groups underwent a 12-week treatment course.

[0076] 2.3 Evaluation Indicators

[0077] 2.3.1 Traditional Chinese Medicine Syndrome Score: This score was formulated with reference to the "Guiding Principles for Clinical Research of New Traditional Chinese Medicine Drugs (Trial Implementation)". The main observed symptoms were: fatigue, shortness of breath, reluctance to speak, frequent urination, soreness and weakness of the lower back and knees, edema, heaviness in the limbs, dull complexion, and rough, dry skin. The above symptoms were divided into four levels according to severity: 0 points for no symptoms, 1 point for mild symptoms, 2 points for moderate symptoms, and 3 points for severe symptoms. Scores were recorded once before and once after treatment.

[0078] 2.3.2 Laboratory Indicators

[0079] The following indicators were measured and recorded before and after treatment in both groups of patients:

[0080] A. Efficacy indicators

[0081] 1) Primary endpoint indicator: UACR.

[0082] 2) Secondary endpoint indicator: eGFR.

[0083] 3) Other efficacy indicators: 24hUP, Uα1-MG, UNAG, FPG, 2hPG, HbA1c, TC, TG, HDL, LDL, NLR, hs-CRP, PA, Alb, Hcy.

[0084] B. Safety indicators: complete blood count, urinalysis, stool analysis, liver function tests, blood electrolytes, and electrocardiogram.

[0085] 2.3.3 Clinical efficacy

[0086] Clinical efficacy was observed using UACR as the primary endpoint and eGFR as the secondary endpoint. The treatment followed the "Diagnostic, Differential Diagnosis and Efficacy Evaluation Criteria for Diabetic Nephropathy (Trial Version)".

[18] The following clinical efficacy criteria are proposed: Marked effect: UACR decreases by more than 1 / 2 or returns to normal; Effective: UACR decreases by less than 1 / 2 compared to before treatment; Ineffective: UACR remains unchanged or increases compared to before treatment.

[0087] 2.3.4 Therapeutic Effects of Traditional Chinese Medicine Syndromes

[0088] Referencing the "Guiding Principles for Clinical Research of New Traditional Chinese Medicine Drugs (Trial Implementation)"

[17] The proposed treatment criteria are as follows: Clinical control: TCM clinical symptoms and signs disappear or basically disappear, with a syndrome efficacy rate ≥90%; Marked effect: TCM clinical symptoms and signs significantly improve, with a syndrome efficacy rate ≥70% (<90%); Effective: TCM clinical symptoms and signs improve, with a syndrome efficacy rate ≥30% (<70%); Ineffective: TCM clinical symptoms and signs show no significant improvement, or even worsen, with a syndrome efficacy rate <30%. (Syndrome efficacy rate (Nimodipine method) = [(Total score before treatment - Total score after treatment) ÷ Total score before treatment] × 100%.)

[0089] 2.4 Statistical Methods

[0090] SPSS 28.0 software was used for statistical processing. Normally distributed measurement data were expressed as mean ± standard deviation. For non-normally distributed continuous data, the median (interquartile range) (M(IQR)) is used; for categorical data, the percentage is used. Normally distributed continuous data are analyzed using t-tests; independent samples t-tests are used for comparisons between groups; paired t-tests are used for comparisons before and after treatment within groups. Non-normally distributed or unequally variably ...

[0091] 3. Results

[0092] 3.1 Baseline Data Balance Analysis

[0093] A total of 104 patients were enrolled in this study and were randomly divided into a treatment group (n=52) and a control group (n=52) using a random number table. The treatment group consisted of 29 males and 23 females, aged 24-79 years. The duration of diabetes mellitus (DM) ranged from 3-33 years, and the duration of disseminated intravascular coagulation (DKD) ranged from 4 months to 10 years. There were 33 cases in G1 stage, 11 cases in G2 stage, 8 cases in G3a stage, 39 cases in A2 stage, and 13 cases in A3 stage. The active ingredient creatinine (UACR) ranged from 30.74-2894.99 mg / g in A2 stage and 309.50-2894.99 mg / g in A3 stage. The 24-hour uptake dose (24hUP) ranged from 0.02-2.48 g in A2 stage and 0.02-0.57 g in A3 stage. The eGFR ranged from 49.97-134.16 ml / min / 1.73 mcg. 2 HbA1c 5.5-7.6%, total TCM syndrome score 9-19.

[0094] The control group consisted of 30 males and 22 females, aged 29-79 years. The duration of diabetes mellitus (DM) ranged from 2 to 32 years, and the duration of disseminated intravascular coagulation (DKD) ranged from 3 months to 10 years. There were 32 cases in G1 stage, 15 cases in G2 stage, 5 cases in G3a stage, 41 cases in A2 stage, and 11 cases in A3 stage. The active antimicrobial clotting factor (UACR) ranged from 31.17 to 3122.02 mg / g in A2 stage and 373.17 to 3122.02 mg / g in A3 stage. The 24-hour uptake dose (24hUP) ranged from 0.01 to 2.54 g in A2 stage and 0.01 to 0.61 g in A3 stage. The eGFR ranged from 47.81 to 156.93 ml / min / 1.73 mcg. 2 HbA1c 5.5-7.4%, total TCM syndrome score 8-18 points.

[0095] There were no statistically significant differences between the two groups in terms of general characteristics, including gender, age, duration of diabetes mellitus (DM), duration of diabetes mellitus (DKD), stage of chronic kernicterus (CKD), stage of albuminuria, UACR, 24hUP, eGFR, HbA1c, and total TCM syndrome score (P > 0.05), indicating that the baseline data of the two groups were well balanced and comparable.

[0096] 3.2 Efficacy Analysis

[0097] 3.2.1 Analysis of the efficacy of the primary endpoint

[0098] Before intervention, there was no statistically significant difference in UACR between the two groups (P > 0.05), making them comparable. After intervention, the UACR in the treatment group decreased significantly compared to before treatment (P < 0.001), while the UACR in the control group decreased compared to before treatment (P < 0.05). The UACR in both groups did not follow a normal distribution as determined by normality tests. The Scheirer-Ray-Hare test was implemented in R language to replace the nonparametric test in the two-way ANOVA. The results showed a statistically significant difference in the main effect of UACR between the two groups (P < 0.01), indicating that the UACR treatment in the treatment group was more effective than that in the control group.

[0099] Statistical analysis was performed on patients according to albuminuria stage. Before intervention, there was no statistically significant difference in UACR between the two groups of stage A2 patients (P>0.05), making them comparable. After intervention, the UACR of stage A2 patients in the treatment group decreased significantly compared to before treatment (P<0.001), while the UACR of stage A2 patients in the control group decreased compared to before treatment (P<0.05). At 4 and 8 weeks of intervention, the UACR of stage A2 patients in the treatment group was significantly lower than that in the control group (P<0.05). Scheirer-Ray-Hare test results showed that there were statistically significant differences in the main effect of UACR time and the main effect between groups between stage A2 patients (P<0.05, P<0.001), suggesting that the efficacy of UACR in stage A2 patients is related to the treatment duration, and the UACR efficacy in the treatment group was better than that in the control group. Before intervention, there was no statistically significant difference in UACR between the two groups of stage A3 patients (P>0.05), making them comparable. After intervention, the UACR of patients in stage A3 of the treatment group decreased significantly compared with that before treatment (P < 0.01), while that of patients in stage A3 of the control group decreased significantly compared with that before treatment (P < 0.05). Scheirer-Ray-Hare test results showed that the main effect of UACR between the two groups of stage A3 patients was statistically significant (P < 0.05), suggesting that the UACR efficacy of the treatment group in stage A3 patients was superior to that of the control group.

[0100] The above results suggest that the efficacy of UACR in DKD stage 2 patients is related to the treatment duration, and that the spleen-tonifying, kidney-nourishing, and blood-activating formula can optimize the therapeutic effect of empagliflozin on UACR in DKD patients. The results are shown in Tables 1, 2, 3, 4, 5, and 6.

[0101] Table 1 Comparison of UACR (mg / g) between the two groups before and after treatment (M(IQR))

[0102]

[0103] Note: *Intra-group comparison P < 0.05, ***Intra-group comparison P < 0.001, UACR normal reference range: 0-30 mg / g

[0104] Table 2 Comparison of UACR (mg / g) intervention between the two groups of patients based on grouping and treatment cycle (M(IQR))

[0105]

[0106] Note: ▲▲ Intergroup comparison P < 0.01, UACR normal reference value: 0-30 mg / g

[0107] Table 3 Comparison of UACR (mg / g) between the two groups of A2 stage patients before and after treatment ( M(IQR))

[0108]

[0109] Note: *Intra-group comparison P < 0.05, ***Intra-group comparison P < 0.001, UACR normal reference range: 0-30m

[0110] Table 4 Comparison of UACR (mg / g) intervention between the two groups of A2 stage patients based on grouping and treatment cycle ( M(IQR))

[0111]

[0112]

[0113] Note: ▲ Intergroup comparison P < 0.05, ▲▲▲ Intergroup comparison P < 0.001, # Time comparison P < 0.05, Normal reference value for UACR: 0-30 mg / g. Table 5 Comparison of UACR (mg / g) between the two groups of A3 stage patients before and after treatment. M(IQR))

[0114]

[0115] Note: *Intra-group comparison P < 0.05, **Intra-group comparison P < 0.01, UACR normal reference range: 0-30 mg / g

[0116] Table 6 Comparison of UACR (mg / g) intervention between the two groups of A3 stage patients based on grouping and treatment cycle ( M(IQR))

[0117]

[0118] Note: ▲ Intergroup comparison P < 0.05, UACR normal reference value: 0-30 mg / g

[0119] 3.2.2 Clinical efficacy analysis

[0120] Clinical efficacy was evaluated using UACR as the primary endpoint. After intervention, 19 patients in the treatment group showed significant improvement, and 15 showed improvement, with a total effective rate of 69.39%. In the control group, 9 patients showed significant improvement, and 17 showed improvement, with a total effective rate of 55.32%. The clinical efficacy of the treatment group was superior to that of the control group (P < 0.05). The results are shown in Table 7.

[0121] Table 7 Comparison of clinical efficacy between the two groups after treatment (cases, %)

[0122]

[0123] Note: Overall effective rate = (significantly effective + effective) / total number of cases; ▲ P < 0.05 for intergroup comparison.

[0124] 3.2.3 Secondary endpoint efficacy analysis

[0125] Before intervention, there was no statistically significant difference in eGFR between the two groups (P > 0.05), making them comparable. After intervention, the eGFR in the treatment group increased compared to before treatment (P < 0.05), while there was no statistically significant difference in eGFR in the control group compared to before treatment (P > 0.05), suggesting that the Jianpi Yishen Qinghua Heluo formula can effectively improve renal function in patients with DKD. Results are shown in Table 8.

[0126] Table 8. eGFR (ml / min / 1.73m) of patients in the two groups before and after treatment. 2 )Compare

[0127]

[0128] Note: *Intragroup comparison P < 0.05, normal reference value for eGFR: > 90 ml / min / 1.73 m 2

[0129] 3.2.4 Analysis of other efficacy indicators

[0130] 1.24hUP efficacy analysis

[0131] Before intervention, there was no statistically significant difference in 24-hour uptake between the two groups (P > 0.05), making them comparable. After intervention, the 24-hour uptake in the treatment group was significantly lower than before treatment (P < 0.001), while the 24-hour uptake in the control group was also lower than before treatment (P < 0.05).

[0132] Statistical analysis was performed on patients according to albuminuria stages. Before intervention, there was no statistically significant difference in 24-hour albumin uptake (24hUP) between the two groups of stage A2 patients (P > 0.05), making them comparable. After intervention, the 24hUP of stage A2 patients in the treatment group was significantly lower than before treatment (P < 0.01), while there was no statistically significant difference in 24hUP of stage A2 patients in the control group (P > 0.05). Before intervention, there was no statistically significant difference in 24hUP of stage A3 patients between the two groups (P > 0.05), making them comparable. After intervention, the 24hUP of stage A3 patients in the treatment group was significantly lower than before treatment (P < 0.01), while there was also a decrease in 24hUP of stage A3 patients in the control group (P < 0.05).

[0133] The results suggest that the combination of Jianpi Yishen Qinghua Heluofang and empagliflozin can effectively reduce the 24-hour uptake (UP) in patients with DKD, and that Jianpi Yishen Qinghua Heluofang can enhance the effect of empagliflozin in reducing the 24-hour uptake (UP) in patients with DKD stage 2. The results are shown in Tables 9 and 10.

[0134] Table 9 Comparison of 24h UP(g) between the two groups before and after treatment (M(IQR))

[0135]

[0136] Note: *Intra-group comparison P < 0.05, ***Intra-group comparison P < 0.001, Normal reference range for 24h UP: 0-0.15g

[0137] Table 10 Comparison of 24hUP(g) stages between the two groups of patients before and after treatment M(IQR))

[0138]

[0139] Note: *Intra-group comparison P < 0.05, **Intra-group comparison P < 0.01, Normal reference range for 24h UP: 0-0.15g

[0140] 2. Analysis of the efficacy of renal tubular function (Uα1-MG, UNAG)

[0141] 1) Analysis of the efficacy of Uα1-MG

[0142] Before intervention, there was no statistically significant difference in Uα1-MG between the two groups (P > 0.05), making them comparable. After intervention, Uα1-MG in the treatment group was significantly lower than before treatment (P < 0.01), while there was no statistically significant difference in Uα1-MG in the control group (P > 0.05). This suggests that the Jianpi Yishen Qinghua Heluo formula can effectively reduce Uα1-MG and improve renal tubular function in patients with DKD. The results are shown in Table 11.

[0143] Table 11 Comparison of Uα1-MG (mg / L) between the two groups of patients before and after treatment

[0144]

[0145]

[0146] Note: **Intragroup comparison P < 0.01, normal reference range for Uα1-MG: 0-12.0 mg / L

[0147] 2) UNAG efficacy analysis

[0148] Before intervention, there was no statistically significant difference in UNAG between the two groups (P > 0.05), making them comparable. After intervention, UNAG in the treatment group was significantly lower than before treatment (P < 0.01), while there was no statistically significant difference in UNAG in the control group (P > 0.05). This suggests that the Jianpi Yishen Qinghua Heluo formula can effectively reduce UNAG and improve renal tubular function in patients with DKD. The results are shown in Table 12.

[0149] Table 12 Comparison of UNAG (mg / L) between the two groups of patients before and after treatment

[0150]

[0151] Note: **Intragroup comparison P < 0.01, UNAG normal reference range: 0-11.5 mg / L

[0152] 3. Analysis of the therapeutic effects of glucose metabolism (FPG, 2hPG, HbA1c)

[0153] Before intervention, there were no statistically significant differences in FPG, 2hPG, and HbA1c between the two groups (P > 0.05), making them comparable. After intervention, FPG, 2hPG, and HbA1c in the treatment group were significantly lower than before treatment (P < 0.001), while FPG in the control group was significantly lower than before treatment (P < 0.01), and 2hPG and HbA1c were significantly lower than before treatment (P < 0.001). FPG, 2hPG, and HbA1c in the treatment group were all lower than those in the control group at the same time point (P < 0.05), suggesting that the Jianpi Yishen Qinghua Heluo formula can effectively improve glucose metabolism in patients with DKD. Results are shown in Table 13.

[0154] Table 13 Comparison of FPG, 2hPG, and HbA1c between the two groups before and after treatment ( M(IQR))

[0155]

[0156]

[0157] Note: **Within-group comparison P < 0.01, ***Within-group comparison P < 0.001, ▲Between-group comparison P < 0.05, FPG normal reference value:

[0158] 4.3-5.9 mmol / L, 2hPG normal reference range: <7.8 mmol / L, HbA1c normal reference range: 3.6-6.0%

[0159] 4. Analysis of the therapeutic effects of lipid metabolism (TC, TG, HDL, LDL)

[0160] Before intervention, there were no statistically significant differences in TC, TG, HDL, and LDL between the two groups (P > 0.05), making them comparable. After intervention, TC, TG, and LDL in the treatment group were significantly lower than before treatment (P < 0.001), while HDL showed no statistically significant difference (P > 0.05). In the control group, TC and LDL were lower than before treatment (P < 0.05), while TG and HDL showed no statistically significant difference (P > 0.05). This suggests that the Jianpi Yishen Qinghua and Luofang formula has a positive effect on improving lipid metabolism in DKD patients, and can effectively enhance the efficacy of empagliflozin in reducing TG in DKD patients. The results are shown in Table 14.

[0161] Table 14 Comparison of TC, TG, HDL, and LDL levels between the two groups before and after treatment ( M(IQR))

[0162]

[0163]

[0164] Note: *Intra-group comparison P < 0.05, ***Intra-group comparison P < 0.001, TC normal reference range: 0-5.18 mmol / L, TG normal reference range: 0-1.70 mmol / L, HDL normal reference range: 1.16-1.42 mmol / L, LDL normal reference range: 0-3.37 mmol / L

[0165] 5. Analysis of the efficacy of inflammatory markers (NLR, hs-CRP)

[0166] Before intervention, there were no statistically significant differences in NLR and hs-CRP between the two groups (P > 0.05), making them comparable. After intervention, NLR and hs-CRP in the treatment group decreased compared to before treatment (P < 0.05), while there were no statistically significant differences in NLR and hs-CRP in the control group compared to before treatment (P > 0.05). This suggests that the Jianpi Yishen Qinghua Heluo formula can effectively improve the inflammatory state in DKD patients. The results are shown in Table 15.

[0167] Table 15 Comparison of NLR and hs-CRP levels between the two groups before and after treatment ( M(IQR))

[0168]

[0169]

[0170] Note: *Intragroup comparison P < 0.05, normal reference range for hs-CRP: 0-0.6 mg / dl

[0171] 6. Serum protein (PA, Alb) efficacy analysis

[0172] Before intervention, there were no statistically significant differences in serum PA and Alb levels between the two groups (P > 0.05), making them comparable. After intervention, PA and Alb levels in the treatment group were significantly higher than before treatment (P < 0.001), while there were no statistically significant differences in PA and Alb levels in the control group (P > 0.05). Alb levels in the treatment group were higher than those in the control group at the same time point (P < 0.05), suggesting that the Jianpi Yishen Qinghua Heluo formula can effectively improve serum PA and Alb levels in DKD patients, with a more significant intervention effect on Alb. The results are shown in Table 16.

[0173] Table 16 Comparison of PA and Alb levels between the two groups before and after treatment.

[0174]

[0175] Note: *** Within-group comparison P < 0.001, ▲ Between-group comparison P < 0.05, PA normal reference range: 200-400 mg / L, Alb normal reference range: 40.0-55.0 g / L

[0176] 7. Analysis of Hcy efficacy

[0177] Before intervention, there was no statistically significant difference in serum Hcy levels between the two groups (P > 0.05), making them comparable. After intervention, serum Hcy levels in the treatment group decreased compared to before treatment (P < 0.05), while there was no statistically significant difference in serum Hcy levels in the control group compared to before treatment (P > 0.05), suggesting that the Jianpi Yishen Qinghua Heluo formula can effectively reduce serum Hcy levels in DKD patients. The results are shown in Table 17.

[0178] Table 17 Comparison of Hcy (μmol / L) between the two groups before and after treatment (M(IQR))

[0179]

[0180] Note: *Intragroup comparison P < 0.05, normal reference range for Hcy: 0-15.0 μmol / L

[0181] 5.2.5 Analysis of the efficacy of traditional Chinese medicine

[0182] 1. Analysis of the therapeutic effects of TCM syndromes

[0183] After intervention, in the treatment group, 1 case of clinical control of TCM syndrome was achieved, 13 cases showed significant improvement, and 30 cases showed improvement, with a total effective rate of 89.80%. In the control group, 0 cases showed clinical control, 0 cases showed significant improvement, and 18 cases showed improvement, with a total effective rate of 38.30%. The TCM syndrome efficacy in the treatment group was significantly better than that in the control group (P < 0.001). The results are shown in Table 18.

[0184] Table 18 Comparison of TCM syndrome efficacy between the two groups of patients after treatment (cases, %)

[0185]

[0186] Note: Overall effective rate = (clinical control + significant effect + effective) / total number of cases, ▲▲▲ intergroup comparison P < 0.001

[0187] 2. Analysis of the therapeutic effects of single syndromes in Traditional Chinese Medicine

[0188] After intervention, the therapeutic effects of each TCM syndrome in the treatment group were significantly better than those in the control group (P < 0.001), indicating that the spleen-tonifying, kidney-tonifying, and blood-activating formulas significantly improved the clinical TCM syndromes of patients with spleen-kidney qi deficiency and damp-heat stagnation type DKD. These formulas can comprehensively improve symptoms such as fatigue, shortness of breath, frequent urination, lower back and knee weakness, edema, heaviness in the limbs, dull complexion, and rough skin in patients with spleen-kidney qi deficiency and damp-heat stagnation type DKD, thus improving the quality of life of DKD patients. Results are shown in Table 19.

[0189] Table 19 Comparison of the efficacy of single TCM syndromes in the two groups of patients after treatment (cases, %)

[0190]

[0191]

[0192]

[0193] Note: Overall effective rate = (clinical control + significant effect + effective) / total number of cases, ▲▲▲ intergroup comparison P < 0.001

[0194] 3.2.6 Security Analysis

[0195] No serious adverse events occurred in either group during treatment. One patient in the treatment group and three patients in the control group withdrew from the trial due to urinary tract infections, which was considered to be related to adverse reactions to empagliflozin. The results of this study show that the Jianpi Yishen Qinghua Heluo formula had no adverse effects on patients' liver and kidney function, electrolytes, or complete blood count.

[0196] Example 5: Animal Model Experiment

[0197] 1. Experimental Methods: Forty healthy male SD rats of SFP grade were randomly divided into four groups: normal group, model group, empagliflozin group, Jianpi Yishen Qinghua Heluofang group, and Jianpi Yishen Qinghua Heluofang combined with empagliflozin group, with eight rats in each group. A DKD rat model was established using a high-sugar, high-fat diet (20.0% sucrose, 10.0% lard, 2.5% cholesterol, 1.0% bile salts, 66.5% conventional feed) combined with a low-dose (35 mg / kg) STZ injection. The normal group and model group were administered distilled water by gavage once daily, while the other three groups were administered the corresponding drugs (6.3 times the human equivalent dose) by gavage once daily, at a dose of 4 ml / kg / day, for 8 weeks. Urine samples were collected from rats in metabolic cages after 24 hours to detect 24-hour urinary output (UP), UACR, Uα1-MG, Uβ2-MG, and UNAG. Blood samples were collected from the abdominal aorta to detect serum SCysC, Scr, BUN, SUA, NLR, hs-CRP, FPG, TC, TG, HDL, LDL, and Hcy. Kidney specimens were stained with HE, PAS, and Masson staining to observe the pathological condition of kidney tissue in each group of rats. The gene expression of NF-κB, TNF-α, IL-8, and MCP-1 in the kidney tissue of each group of rats was detected by qRT-PCR. The protein expression of AGEs, RAGE, NF-κB, TNF-α, IL-8, MCP-1, and VEGF in the kidney tissue of each group of rats was detected by Western blotting.

[0198] 2. Experimental Results

[0199] Regarding body weight and kidney coefficient, Jianpi Yishen Qinghua Heluofang could increase the body weight of DKD rats (P<0.05 compared with the model group) and decrease the kidney coefficient of DKD rats (P<0.01 compared with the model group). The combined use of Jianpi Yishen Qinghua Heluofang and Jianpi Yishen Qinghua Heluofang was more effective than either drug alone (P<0.05 or P<0.001 compared with the single drug group). Regarding 24-hour urine output, both Jianpi Yishen Qinghua Heluofang and Jianpi Yishen Qinghua Heluofang were effective in improving polyuria symptoms in DKD rats (P<0.001 or P<0.01 compared with the model group). The combined use of Jianpi Yishen Qinghua Heluofang and Jianpi Yishen Qinghua Heluofang was more effective than either drug alone (P<0.05 compared with the single drug group).

[0200] Regarding proteinuria, empagliflozin alone or in combination with Jianpi Yishen Qinghua and Luofang both reduced UACR (P < 0.05 or P < 0.01 compared with the model group) and 24hUP (P < 0.01 or P < 0.001 compared with the model group) in DKD rats. The combination of the two drugs was more effective than either drug alone in reducing UACR and 24hUP (UACR: P < 0.05 compared with Jianpi Yishen Qinghua and Luofang group, P > 0.05 compared with empagliflozin group; 24hUP: P < 0.001 compared with Jianpi Yishen Qinghua and Luofang group, P < 0.05 compared with empagliflozin group). The combination of the two drugs also reduced UNAG in DKD rats (P > 0.05 compared with the normal group, P < 0.05 compared with the model group). See Table 20 for details.

[0201] Table 20 Comparison of UACR (mg / g), 24hUP (mg), and UNAG (mg / L) among rats in each group after intervention. M(IQR))

[0202]

[0203] Note: * P < 0.05 compared with the normal group, ** P < 0.01 compared with the normal group, *** P < 0.001 compared with the normal group, ▲ P < 0.05 compared with the model group, ▲▲ P < 0.01 compared with the model group, ▲▲▲ P < 0.001 compared with the model group, # P < 0.05 compared with the empagliflozin group, ## P < 0.01 compared with the empagliflozin group, △ P < 0.05 compared with the Jianpi Yishen Qinghua Heluo Fang group, △△△ P < 0.001 compared with the Jianpi Yishen Qinghua Heluo Fang group.

[0204] Regarding renal function, Jianpi Yishen Qinghua and Luofang could reduce Scr, BUN, and SUA in DKD rats (P < 0.05 compared with the model group). The combination of the two drugs could reduce SCysc in DKD rats (P < 0.05 compared with the model group), and the efficacy was superior to empagliflozin alone (P < 0.05 compared with the empagliflozin group). Regarding inflammation, both empagliflozin and Jianpi Yishen Qinghua and Luofang could reduce hs-CRP in DKD rats (P > 0.05 compared with the normal group). The combination of the two drugs was more effective in reducing NLR in DKD rats than either drug alone (P < 0.01 compared with the model group). Regarding glucose metabolism, Jianpi Yishen Qinghua and Luofang could reduce FPG in DKD rats (P < 0.01 compared with the model group). (P < 0.05); Regarding lipid metabolism, Jianpi Yishen Qinghua Heluo Fang can reduce TC, TG, and LDL in DKD rats (P < 0.05, P < 0.01, or P < 0.001 compared with the model group), and its efficacy in reducing TG and LDL is better than that of empagliflozin (P < 0.05 compared with the empagliflozin group). Combined use with empagliflozin can enhance the efficacy of empagliflozin in reducing TC, TG, and LDL (P < 0.05 or P < 0.01 compared with the empagliflozin group); Regarding risk factors, Jianpi Yishen Qinghua Heluo Fang can reduce Hcy in DKD rats (P < 0.05 compared with the model group), and the combination of the two drugs can improve the efficacy of empagliflozin in reducing Hcy (P < 0.05 compared with the empagliflozin group). Pathological staining results showed that Jianpi Yishen Qinghua and Luofang could reduce the proliferation of glomerular basement membrane and mesangial matrix in DKD rats, and reduce renal interstitial inflammatory cell infiltration, renal tubular lesions, and renal interstitial fibrosis (PAS positive rate and Masson positive rate compared with the model group, P < 0.001). qRT-PCR results showed that Jianpi Yishen Qinghua and Luofang could downregulate the gene expression of NF-κB, TNF-α, IL-8, and MCP-1 in the renal tissue of DKD rats (compared with the model group, P < 0.01). The combination of the two drugs enhanced the downregulation effect of Jianpi Yishen Qinghua and Luofang on the gene expression of NF-κB, TNF-α, IL-8, and MCP-1 (compared with the Jianpi Yishen Qinghua and Luofang group, P < 0.001 or P < 0.01), and enhanced the downregulation effect of empagliflozin on the gene expression of IL-8 and MCP-1 in the renal tissue of DKD rats (compared with the normal group, P > 0.05). Western blot results showed that the Jianpi Yishen Qinghua Heluo formula could downregulate the protein expression of AGEs, RAGE, NF-κB, TNF-α, IL-8, and VEGF in the kidney tissue of DKD rats (P < 0.001 or P < 0.01 compared with the model group). When used in combination with empagliflozin, it could downregulate the protein expression of MCP-1 in the kidney tissue of DKD rats (P < 0.01 compared with the model group). The combined use of the two drugs was more effective than the use of either drug alone in downregulating the protein expression of AGEs, RAGE, NF-κB, TNF-α, IL-8, and VEGF (P < 0.001 or P < 0.01 compared with the single drug group).

[0205] 3. Experimental Conclusions

[0206] The spleen-tonifying, kidney-nourishing, and blood-activating formula can effectively alleviate pathological symptoms such as emaciation and polyuria in DKD rats, improve inflammation and glucose-lipid metabolism imbalance, reduce risk factors for renal vascular lesions in DKD rats, protect renal function, and enhance the proteinuria-reducing effect of empagliflozin. One of its kidney-protective mechanisms may be through regulating related inflammatory factors via the AGEs-RAGE signaling pathway, inhibiting the inflammatory response in DKD, improving renal vascular endothelial function, alleviating renal lesions, and reducing proteinuria in DKD rats.

[0207] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. A pharmaceutical composition for treating diabetic nephropathy, characterized in that, It consists of a traditional Chinese medicine composition and therapeutic agents used in combination with it. The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 10-60 parts Astragalus membranaceus, 5-30 parts Polygonatum sibiricum, 5-30 parts Dioscorea opposita, 5-30 parts Cornus officinalis, 3-15 parts Coptis chinensis, 5-30 parts Euonymus alatus, 5-30 parts Arctium lappa, and 3-15 parts Hirudo medicinalis. The therapeutic agent is empagliflozin.

2. The pharmaceutical composition for treating diabetic nephropathy according to claim 1, characterized in that, The traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 30 parts of raw Astragalus membranaceus, 15 parts of Polygonatum sibiricum, 15 parts of Dioscorea opposita, 15 parts of Cornus officinalis, 6 parts of Coptis chinensis, 15 parts of Euonymus alatus, 15 parts of Arctium lappa, and 6 parts of Hirudo medicinalis.

3. The pharmaceutical composition for treating diabetic nephropathy according to claim 1, characterized in that, Astragalus membranaceus is the principal herb, Polygonatum sibiricum, Dioscorea opposita, and Cornus officinalis are the assistant herbs, Coptis chinensis and Euonymus alatus are the adjuvant herbs, and Arctium lappa and Hirudo medicinalis are the guiding herbs.

4. The pharmaceutical composition for treating diabetic nephropathy according to claim 1, characterized in that, The formulation of the traditional Chinese medicine composition is prepared into a commonly used dosage form in pharmaceuticals using conventional preparation methods in the field.

5. The pharmaceutical composition for treating diabetic nephropathy according to claim 4, characterized in that, The traditional Chinese medicine composition is prepared as a decoction, granules, tablets, capsules, oral liquid, mixture, or syrup.

6. The pharmaceutical composition for treating diabetic nephropathy according to claim 4, characterized in that, The preparation method of the traditional Chinese medicine composition includes the following steps: (A) Preparation of the spleen-tonifying, kidney-tonifying, clearing and purging decoction using the traditional decoction method: Weigh the Chinese medicinal herbs according to the formula ratio, add drinking water to the water level 2 cm above the herbs, soak for 30 minutes, heat over high heat until the decoction boils, then reduce to low heat and continue to simmer for 30 minutes. Pour out the decoction, add water to the dregs again, decoct using the same method, and pour out the decoction. Combine the two decoctions, filter and further concentrate to obtain the water extract of the Chinese medicinal composition. (B) Prepared by adding a pharmaceutically acceptable carrier to the aqueous extract using conventional methods in the art.

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