Use of sanguirhizine in preparation of drugs for preventing and treating renal fibrosis

Abstract

The application discloses application of anemarrhenanine in preparation of a drug for preventing and treating renal fibrosis. The application first proposes and verifies that anemarrhenanine can effectively regulate the M1 / M2 macrophage ratio in kidney tissue, improve the inflammatory environment, promote tubular epithelial cell ferroptosis, and down-regulate the expression of fibrosis-related proteins, thereby realizing the effects of protecting kidney function and relieving the progression of renal fibrosis, and can be used for research and development of a clinical treatment drug for renal fibrosis-related diseases.

Description

Technical Field

[0001] This invention relates to the field of natural product chemistry, and in particular to the application of Aconitum carmichaelii in the preparation of drugs for the prevention and treatment of renal fibrosis. Background Technology

[0002] Chronic kidney disease (CKD) is an irreversible disease characterized by progressive loss of kidney function. Currently, it affects approximately 850 million people worldwide, with a prevalence of about 10%–15%, making it another major public health problem after cardiovascular disease and cancer. Early symptoms of CKD are insidious, and awareness is extremely low. Many patients suffer kidney damage unknowingly, potentially progressing to end-stage renal disease and requiring dialysis or kidney transplantation to sustain life.

[0003] Renal fibrosis is a common pathological pathway in the progression of various chronic kidney diseases to end-stage renal failure. Essentially, it is a highly dynamic multicellular process—initial tissue damage triggers an inflammatory cascade response. Infiltrating inflammatory cells, especially macrophages, exhibit M1 (pro-inflammatory) or M2 (pro-repair) phenotypes in the microenvironment. The persistent imbalance of inflammatory signals maintains myofibroblast activation through pathways such as TGF-β / Smad and NF-κB, leading to excessive extracellular matrix deposition, renal tubular atrophy, and microvascular depletion, ultimately forming irreversible scarring. Currently, commonly used clinical treatments such as ACE inhibitors and SGLT2 inhibitors can only slow the progression of fibrosis; they cannot reverse the existing pathological changes.

[0004] Chasmanine (Cha) is a C19-diterpenoid alkaloid, mainly isolated from the roots of plants in the genus Aconitum. The role of Chasmanine in the treatment of renal fibrosis is currently unreported and requires further investigation. Summary of the Invention

[0005] Purpose of the invention: The purpose of this invention is to provide a method for treating renal fibrosis by synergistically regulating the ratio of M1 / M2 macrophages in kidney tissue, improving the inflammatory environment, promoting ferroptosis of renal tubular epithelial cells, and downregulating fibrosis-related proteins, thereby protecting kidney function and alleviating the progression of renal fibrosis. Based on this, the invention also provides its application in the preparation of drugs for the prevention and treatment of renal fibrosis.

[0006] Technical solution: The application of Aconitum carmichaelii described in this invention in the preparation of drugs for the prevention and treatment of renal fibrosis.

[0007] Preferably, the CAS number of the aconitine plantarum is 5066-78-4.

[0008] Preferably, the application is in the preparation of a medicament for protecting kidney function and / or inhibiting renal collagen fiber deposition.

[0009] Preferably, the application is in the preparation of a drug that inhibits the differentiation of macrophage M1 type.

[0010] Preferably, the application is in the preparation of a drug that promotes the differentiation of macrophages M2 type.

[0011] Preferably, the application is in the preparation of a drug that inhibits abnormal activation of renal epithelial cells.

[0012] Preferably, the drug contains aconitine or its pharmaceutically acceptable salts, solvates, or hydrates as active ingredients.

[0013] Preferably, the drug further contains pharmaceutically acceptable excipients; more preferably, the pharmaceutically acceptable excipients include excipients selected from one or more of diluents, lubricants, flow aids, wetting agents, emulsifiers, or pH buffers.

[0014] Preferably, the dosage form of the drug includes tablets, capsules, granules, oral liquids, syrups, powders, microcapsules, injections, powder injections, infusions, suspensions, ointments, creams, gels, sprays, eye drops, ear drops, nasal drops, patches, lotions, suppositories, dressings, nebulizing solutions, film-forming agents, implants, microemulsions, liposomes, nanoparticles, oral instant films, sponges, and capsules.

[0015] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: The present invention proposes and verifies for the first time that Aconitum carmichaelii can effectively regulate the ratio of M1 / M2 macrophages in kidney tissue, improve the inflammatory environment, promote ferroptosis of renal tubular epithelial cells, and downregulate the expression of fibrosis-related proteins, thereby achieving the effect of protecting kidney function and alleviating the progression of renal fibrosis. It can be used for the research and development of clinical treatment drugs for renal fibrosis-related diseases. Attached Figure Description

[0016] Figure 1 Figure 1 shows the results of renal function index detection in mice with renal fibrosis model after treatment with Aconitum carmichaelii. Figure 2 Representative H&E staining of kidney tissue from a mouse model of renal fibrosis after treatment with Aconitum carmichaelii; Figure 3 A quantitative analysis of immune cell infiltration in the renal tissue of a mouse model of renal fibrosis after treatment with Aconitum carmichaelii; Figure 4 Representative Masson staining images of kidney tissue from a mouse model of renal fibrosis after treatment with Aconitum carmichaelii; Figure 5 A quantitative analysis of fibrin deposition in the renal tissue of a mouse model of renal fibrosis after treatment with Aconitum carmichaelii; Figure 6Representative images of immunohistochemical staining of collagen markers in kidney tissue of mice with renal fibrosis model after treatment with Aconitum carmichaelii; Figure 7 A graph showing the quantitative analysis of collagen markers in the renal tissue of a mouse model of renal fibrosis after treatment with Aconitum carmichaelii. Figure 8 Figure 1 shows the results of measuring cytokine levels in the renal tissue of a mouse model of renal fibrosis after treatment with Aconitum carmichaelii. Figure 9 Figure 1. Flow cytometry results of macrophage typing in renal tissue of mice with renal fibrosis model after treatment with Aconitum carmichaelii; Figure 10 Figure 1 shows the results of measuring the expression levels of JAK / STAT3 pathway-related proteins in the kidney tissue of mice with renal fibrosis model after treatment with Aconitum carmichaelii. Figure 11 Figure 1. Quantitative analysis of the expression levels of JAK / STAT3 pathway-related proteins in the kidney tissue of mice with renal fibrosis model after treatment with Aconitum carmichaelii. Figure 12 Figure 1 shows the NO level determination results in the co-incubation system after treatment with aconitine. Figure 13 Figure showing the results of cytokine level measurement in the co-incubation system after treatment with aconitine in *Platycodon grandiflorus*. Figure 14 The flow cytometry results of M1 typing in RAW264.7 macrophages in the co-incubation system after treatment with Aconitum carmichaelii are shown in the figure. Figure 15 The image shows the flow cytometry results of M2 typing in RAW264.7 macrophages in the co-incubation system after treatment with Aconitum carmichaelii. Figure 16 Figure 1 shows the results of measuring the expression levels of JAK / STAT3 pathway-related proteins in RAW264.7 macrophages in a co-incubation system after treatment with Aconitum carmichaelii. Figure 17 A quantitative analysis of the expression levels of JAK / STAT3 pathway-related proteins in RAW264.7 macrophages after co-incubation with Aconitum carmichaelii treatment. Figure 18 Figure 1 shows the results of measuring the expression levels of collagen markers in HK2 cells of the co-incubation system after treatment with Aconitum carmichaelii. Figure 19 Figure 1. Flow cytometry analysis results of ROS levels in HK2 cells in a co-incubation system after treatment with Aconitum carmichaelii; Figure 20 Flow cytometry results of lipid peroxidation levels in HK2 cells after co-incubation with Aconitum carmichaelii treatment; Figure 21 The image shows the results of ferroptosis marker determination in HK2 cells of the co-incubation system after treatment with Aconitum carmichaelii. Detailed Implementation

[0017] The technical solution of the present invention will be further described below.

[0018] Example 1: In vivo efficacy verification of Chasmanine (Cha) for the prevention and treatment of renal fibrosis 1. Construction and drug treatment of animal models of renal fibrosis A 5 / 6 nephrectomy was performed to establish a model of renal fibrosis in chronic renal failure.

[0019] Male C57BL / 6J mice aged 6-8 weeks were purchased from the Experimental Animal Center of Nantong University and housed under standard laboratory conditions. After a one-week acclimatization period, they were randomly divided into a sham surgery group (n=3) and a 5 / 6 nephrectomy group (n=6). In the 5 / 6 nephrectomy group, mice were anesthetized, and the left kidney was exposed through a 1 cm oblique subcostal incision. Two-thirds of the left kidney tissue (one-third at the upper pole and one-third at the lower pole) was removed. Hemostasis was achieved by applying gelatin sponge for 10 minutes, and the wound was sutured in layers. Seven days later, under the same anesthesia conditions, the right renal vessels and ureter were ligated, and the right kidney was completely removed, resulting in the removal of 5 / 6 of the functional kidney tissue. Mice in the sham surgery group underwent only anesthesia and bilateral kidney exposure, without tissue removal.

[0020] Fourteen days after model establishment, mice in the 5 / 6 nephrectomy group were randomly divided into two groups (n=3 per group): the RF group and the RF+Cha group, and drug treatment was initiated simultaneously. The drugs were administered via gavage once daily. Mice in the RF+Cha group received 20 mg / kg of Cha (purchased from MedChemExpress LLC., catalog number HY-N1946), while mice in the Sham and RF groups received the same volume of saline. This treatment continued for 8 weeks. Subsequently, the mice were euthanized, and kidney tissue was harvested for further analysis.

[0021] 2. Kidney function test Mouse kidney tissue was collected and ground in liquid nitrogen. Pre-cooled physiological saline was added to prepare a tissue homogenate at a ratio of 1 mL per 100 mg of tissue. After centrifugation at 12,000 rpm for 15 min at 4°C, the supernatant was collected. The contents of creatinine, urea nitrogen and uric acid were determined using the Elite creatinine (Cr) colorimetric assay kit (product number E-BC-K188-M), the Elite urea (BUN) colorimetric assay kit (product number E-BC-K183-M) and the Elite uric acid (UA) colorimetric assay kit (product number E-BC-K016-M), respectively.

[0022] The results are as follows Figure 1As shown, renal function was significantly reduced in the RF group mice, while renal function was significantly restored after Cha treatment (RF+Cha group). This initially suggests that Cha can be used as a potential treatment for renal fibrosis.

[0023] 3. Pathological evaluation of kidney tissue 3.1 H&E staining Mouse kidney tissues were fixed in 4% paraformaldehyde solution for 48 h, dehydrated in a gradient manner, embedded in paraffin, and cut into 3 μm sections. After dewaxing and hydration, hematoxylin-eosin (H&E) staining was performed, and the sections were mounted with neutral resin and observed under a microscope to collect images. Subsequently, the infiltration of immune cells in the kidney tissues was quantitatively analyzed.

[0024] The results are as follows Figure 2 , 3 As shown, the RF group mice exhibited glomerular sclerosis, mesangial cell and matrix proliferation, renal tubular atrophy, renal tubular epithelial cell shedding, vacuolar degeneration, tubular dilation, and significant infiltration of inflammatory cells in the renal interstitium. These pathological changes were significantly reduced in the Cha-treated group mice.

[0025] 3.2 Masson staining The preliminary steps were the same as those for H&E staining. In the staining section, the collagen deposits in the liver tissue were stained using the Solarbio Masson trichrome staining kit (catalog number G1340). After mounting with neutral resin, the tissue was observed and images were acquired under a microscope. Subsequently, the area ratio of collagen fibers in the kidney tissue was quantitatively analyzed.

[0026] The results are as follows Figure 4 , 5 As shown, collagen deposition (blue stained area) in the renal interstitium of mice in the RF group was significantly increased, and the positive rate of collagen staining was elevated; after treatment with Cha, collagen deposition was significantly reduced; quantitative analysis showed that compared with mice in the RF group, the collagen-positive area of ​​mice in the Cha-treated group was reduced by 21.5%.

[0027] 3.3 Immunohistochemical staining analysis Mouse kidney tissue was fixed in 4% paraformaldehyde solution for 48 h, dehydrated in a gradient manner, embedded in paraffin, cut into 4 μm sections, and antigen retrieval was performed at 95 °C for 20 min with citrate buffer at pH 6.0. Endogenous peroxidase activity was inhibited with 3% hydrogen peroxide solution for 10 min, followed by blocking with 5% bovine serum albumin solution for 1 h. The tissue was incubated overnight at 4 °C with α-SMA primary antibody (purchased from Proteintech, catalog number 14395-1-AP) diluted 1:10000 or Col-I primary antibody (purchased from Proteintech, catalog number 83752-5-RR) diluted 1:1000. After washing, the sections were incubated with horseradish peroxidase (HRP)-conjugated secondary antibody (Proteintech, catalog number SA00004-2) diluted 1:500 at room temperature for 1 h; signal detection was performed using the Beyotime DAB chromogenic reagent kit (catalog number P0202), and the cell nuclei were counterstained with hematoxylin. Images were acquired using an optical microscope, and relative protein levels were quantified.

[0028] The results are as follows Figure 6 , 7 As shown, the concentrations of extracellular matrix (ECM) components α-SMA and Col-I in the kidney tissue of mice in the RF group were significantly higher than those in the sham group; however, after Cha treatment, the levels of α-SMA and Col-I decreased significantly.

[0029] 4. Measurement of renal tissue cytokine levels Mouse kidney tissue was collected and homogenized in liquid nitrogen. Pre-cooled physiological saline was added to prepare a tissue homogenate at a ratio of 1 mL per 100 mg of tissue. After centrifugation at 12,000 rpm for 15 min at 4°C, the supernatant was collected. The cytokine levels in the kidney tissue were measured using the following kits: Ellete mouse interleukin-1β (IL-1β) enzyme-linked immunosorbent assay kit (Catalog No. E-EL-M0037), Ellete mouse interleukin-6 (IL-6) enzyme-linked immunosorbent assay kit (Catalog No. E-EL-M0044), Ellete mouse tumor necrosis factor-α (TNF-α) enzyme-linked immunosorbent assay kit (Catalog No. E-EL-M3063), and Ellete mouse interleukin-10 (IL-10) enzyme-linked immunosorbent assay kit (Catalog No. E-EL-M0046).

[0030] The results are as follows Figure 8 As shown, the levels of IL-1β, IL-6, and TNF-α in the kidney tissue of mice in the RF group were significantly higher than those in the sham group, while the level of IL-10 was significantly lower; Cha treatment was able to alleviate the progression of renal fibrosis by inhibiting the expression of pro-inflammatory cytokines in mice.

[0031] 5. Analysis of macrophage subtypes in kidney tissue Collected mouse kidney tissue was minced into pieces of 0.5-1 mm. 3 Small pieces of tissue were treated with the Absin mouse kidney tissue dissociation kit (catalog number abs50095) to obtain dissociated kidney tissue cells. After staining with Elite PE Anti-Mouse CD86 Antibody [GL-1] (catalog number E-AB-F0994D) and Elite APC Anti-Mouse CD206 / MMR Antibody [C068C2] (catalog number E-AB-F1135E), flow cytometry was performed to detect and analyze the subtypes of macrophages in the kidney tissue.

[0032] The results are as follows Figure 9 As shown, in the sham group mice, the proportion of M1 macrophages was 72% and the proportion of M2 macrophages was 25%; in the kidney tissue of the RF group mice, the proportion of M1 macrophages was 87% and the proportion of M2 macrophages was 9%; while after Cha treatment, the proportion of M1 macrophages was 56% and the proportion of M2 macrophages was 38%. This indicates that Cha can significantly inhibit the proportion of M1 macrophages in kidney tissue, thereby alleviating the inflammatory mediator environment and achieving the treatment of renal fibrosis.

[0033] 6. Determination of JAK / STAT3 pathway-related proteins in kidney tissue Mouse kidney tissue was collected and ground in liquid nitrogen using a mortar and pestle. For every 100 mg of tissue, 1 mL of RIPA lysis buffer (Catalog No. WB3100) containing 0.1 volume of a mixture of cyprotease inhibitors (P001) and cyprotease inhibitors (P003) was added. Lysis was performed at 4°C for 30 min, followed by centrifugation at 12000 rpm for 30 min. The supernatant was collected and added to 5× Loading buffer (Catalog No. WB2001). The mixture was heated to boiling in a metal bath for 10 min, followed by SDS-PAGE electrophoresis. After transfer and blocking, JAK2 primary antibody (purchased from Proteintech, Catalog No. 17670-1-AP) diluted 1:1000 and p-JAK2 primary antibody (purchased from MedChemExpress) diluted 1:500 were added. LLC., catalog number HY-P80829), STAT3 primary antibody diluted 1:10000 (Proteintech, catalog number 10253-2-AP), p-STAT3 primary antibody diluted 1:2000 (Proteintech, catalog number 80199-2-RR), or GAPDH primary antibody diluted 1:20000 (Proteintech, catalog number 10494-1-AP) were incubated overnight at 4°C. After rinsing, secondary antibody diluted 1:10000 (Proteintech, catalog number SA00001-2) was incubated at room temperature for 2 h. Finally, the chemiluminescence was developed and images were acquired using the NewSymex Ultrasensitive ECL Chemiluminescence Kit (catalog number P10100), and the proportion of phosphorylated proteins was quantitatively analyzed.

[0034] The results are as follows Figure 10 , 11 As shown, the normalized p-JAK2 / JAK2 and p-STAT3 / STAT3 levels in the kidney tissue of RF group mice were 7 times and 4 times higher than those in sham group mice, respectively, while after Cha treatment, they decreased to 2 times and 0.8 times, respectively; that is, compared with sham group mice, the JAK2 / STAT3 pathway was significantly activated in the kidney tissue of RF group mice, while Cha treatment could inhibit the activation of the JAK2 / STAT3 pathway.

[0035] Example 2: In vitro efficacy verification of Aconitum carmichaelii in the prevention and treatment of renal fibrosis 1. Construction and drug treatment of a co-culture system of macrophages and renal epithelial cells HK2 Human renal cortical proximal tubular epithelial cells HK2 at 5×10 4 Cells were seeded at a density of 1.5 × 10⁶ cells / well in the upper chamber of the Transwell (Corning, catalog number 3460) and 1.5 × 10⁶ cells / well in the lower chamber of the corresponding 24-well plate. 5RAW264.7 cells / well were used. The upper chamber used RPMI-1640 medium supplemented with 10% fetal bovine serum; the lower chamber used RPMI-1640 complete medium containing 1‰ DMSO (DMSO group), or RPMI-1640 complete medium containing 1‰ DMSO and 200 ng / mL LPS (purchased from MedChemExpress LLC., catalog number HY-D1056) (LPS group), or RPMI-1640 complete medium containing 1‰ DMSO, 200 ng / mL LPS, and 10 μM Cha (LPS+Cha group). All were cultured in a dedicated incubator at 37°C and 5% CO2 to form a co-culture system. The drug-containing medium was replaced every 24 h for a total of 72 h.

[0036] 2. Determination of nitric oxide (NO) levels in culture supernatant After drug treatment, the cell culture supernatant in the lower chamber was collected, and the NO level was determined using the Elite Nitric Oxide Colorimetric Assay Kit (catalog number E-BC-K135-M).

[0037] The results are as follows Figure 12 As shown, the NO release in the LPS-treated group reached 16.7 μmol / L, indicating that LPS could fully activate macrophages; while the NO release in the LPS+Cha group was only 5.1 μmol / L, indicating that Cha could fully inhibit macrophage activity.

[0038] 3. Measurement of cytokine levels in culture supernatant After drug treatment, the cell culture supernatant from the lower chamber was collected, and the cytokine levels were measured using the Ellete mouse interleukin-6 (IL-6) enzyme-linked immunosorbent assay kit and the Ellete mouse interleukin-10 (IL-10) enzyme-linked immunosorbent assay kit.

[0039] The results are as follows Figure 13 As shown, the level of pro-inflammatory cytokine IL-6 in the LPS-treated group was significantly higher than that in the DMSO group, while the level of IL-10 was significantly lower; Cha treatment was able to downregulate the level of pro-inflammatory cytokines, promote the expression of anti-inflammatory cytokines, and inhibit macrophage activation.

[0040] 4. Macrophage subtype analysis After drug treatment, RAW264.7 cells cultured in the lower chamber were collected and stained with Elite PE Anti-Mouse CD86 Antibody [GL-1] and Elite APC Anti-Mouse CD206 / MMR Antibody [C068C2] before flow cytometry analysis to detect and analyze the RAW264.7 macrophage subtypes.

[0041] The results are as follows Figure 14 , 15 As shown, consistent with the trends observed in in vivo experiments, Cha treatment significantly downregulated the proportion of M1 macrophages in the LPS group while increasing the proportion of M2 macrophages, further regulating the expression levels of inflammatory cytokines, thereby achieving the goal of treating renal fibrosis.

[0042] 5. Assay of JAK / STAT3 pathway-related proteins in macrophages After drug treatment, RAW264.7 cells cultured in the lower chamber were collected and lysed using RIPA lysis buffer containing 0.1 volume of a mixture of cytosine protease inhibitor and cytosine phosphatase inhibitor. The lysate was collected at 4°C for 30 min, centrifuged at 12,000 rpm at 4°C for 30 min, and the supernatant was collected to obtain a whole protein sample. Western blot analysis was performed according to the method described in Example 1. Finally, the sample was developed and images were acquired using a cytosine ultrasensitive ECL chemiluminescence kit, and the proportion of phosphorylated proteins was quantitatively analyzed.

[0043] The results are as follows Figure 16 , 17 As shown, consistent with in vivo experimental trends, Cha can inhibit the activation of the JAK2 / STAT3 pathway induced by LPS treatment.

[0044] 6. Determination of HK2 cell fibrosis marker proteins After drug treatment, HK2 cells cultured in the upper chamber were collected, and total protein samples were extracted according to the aforementioned method. NewSyne 5× Loading buffer was added in proportion, and the mixture was heated in a metal bath to boiling for 10 min before SDS-PAGE electrophoresis. After transfer and blocking, α-SMA primary antibody diluted 1:10000, Col-I primary antibody diluted 1:1000, or GAPDH primary antibody diluted 1:20000 were added and incubated overnight at 4°C. After rinsing, secondary antibody diluted 1:10000 (purchased from Proteintech, catalog number SA00001-2) was added and incubated at room temperature for 2 h. Finally, the cells were developed using the NewSyne ultrasensitive ECL chemiluminescence kit, and images were acquired.

[0045] The results are as follows Figure 18 As shown, the levels of α-SMA and Col-I in HK2 cells of the LPS group were significantly higher than those in the DMSO group, while Cha could significantly downregulate the expression levels of the above two proteins.

[0046] 7. Measurement of reactive oxygen species (ROS) levels in HK2 cells After drug treatment, HK2 cells cultured in the upper chamber were collected, washed with pre-cooled PBS, and stained with a DCFH-DA probe (purchased from MedChemExpress LLC., catalog number HY-D0940) at a final concentration of 5 μM at 37°C for 30 min. The cells were then detected by flow cytometry.

[0047] 8. Measurement of lipid peroxidation level in HK2 cells After drug treatment, HK2 cells cultured in the upper chamber were collected, washed with pre-cooled PBS, and stained with BODIPY581 / 591 C11 probe (purchased from MedChemExpress LLC., catalog number HY-D1301) at 37°C for 30 min, and detected by flow cytometry.

[0048] The results are as follows Figure 19 , 20 As shown, Cha can significantly upregulate ROS and lipid peroxidation levels in HK2 cells, thereby promoting ferroptosis in HK2 cells.

[0049] 9. Determination of HK2 cell ferroptosis marker levels After drug treatment, HK2 cells cultured in the upper chamber were collected, resuspended in PBS, and the cells were lysed. After centrifugation at 1500 × g for 10 min at 4°C, the supernatant was collected. Malondialdehyde (MDA) and ferrous ions (Fe2+) in HK2 cells were detected using the following kits: Elite MDA Enzyme-Linked Immunosorbent Assay Kit (E-EL-0060), 4-Hydroxynonenoic Acid Enzyme-Linked Immunosorbent Assay Kit (E-EL-0128), Total Iron Ion Colorimetric Assay Kit (E-BC-K772-M), and Glutathione (GSH) Enzyme-Linked Immunosorbent Assay Kit (E-EL-0026). 2+ The levels of 4-hydroxynonenal (4-HNE) and glutathione (GSH) The results are as follows Figure 21 As shown, Cha can significantly upregulate intracellular MDA, 4-HNE, and Fe in HK2 cells. 2+ It significantly reduced the level of GSH, while also inhibiting GSH expression and promoting HK2 ferroptosis in renal epithelial cells, ultimately achieving the goal of alleviating renal fibrosis.

Claims

1. The application of Aconitum carmichaelii in the preparation of drugs for the prevention and treatment of renal fibrosis.

2. The application according to claim 1, characterized in that, The CAS number for the aforementioned Aconitum carmichaelii is 5066-78-4.

3. The application according to claim 1, characterized in that, The application is in the preparation of drugs that protect kidney function and / or inhibit renal collagen fiber deposition.

4. The application according to claim 1, characterized in that, The application is in the preparation of drugs that inhibit the differentiation of macrophage M1 type.

5. The application according to claim 1, characterized in that, The application is in the preparation of drugs that promote the differentiation of macrophages M2 type.

6. The application according to claim 1, characterized in that, The application is in the preparation of drugs that inhibit abnormal activation of renal epithelial cells.

7. The application according to claim 1, characterized in that, The drug contains aconitine or its pharmaceutically acceptable salts, solvates, or hydrates as active ingredients.

8. The application according to claim 7, characterized in that, The drug also contains pharmaceutically acceptable excipients.

9. The application according to claim 8, characterized in that, The pharmaceutically acceptable excipients include excipients selected from one or more of diluents, lubricants, flow aids, wetting agents, emulsifiers, or pH buffers.

10. The application according to claim 1, characterized in that, The dosage forms of the drugs include tablets, capsules, granules, oral liquids, syrups, powders, microcapsules, injections, powder injections, infusions, suspensions, ointments, creams, gels, sprays, eye drops, ear drops, nasal drops, patches, lotions, suppositories, dressings, nebulizing solutions, film-forming agents, implants, microemulsions, liposomes, nanoparticles, oral instant films, sponges, and capsules.

Images