Application of sennoside B in preparation of medicine for preventing and treating renal fibrosis caused by urinary tract obstruction

Abstract

The invention relates to the technical field of medicines, discloses application of sennoside B in preparation of medicines for preventing and treating renal fibrosis caused by urinary tract obstruction, and discloses a method for treating renal fibrosis more safely and effectively. According to the method, the natural anti-inflammatory characteristic of sennoside B is utilized, the side effect of the medicine is reduced, and the treatment safety is improved. Besides, the sennoside B is proved to be capable of remarkably reducing the occurrence of mitochondrial autophagy, maintaining the normal function of mitochondria and relieving the occurrence of renal fibrosis, so that a new therapeutic target is provided for treating renal fibrosis, and meanwhile, the sennoside B can be used as a supplement or replacement of an existing therapeutic method, provides a new therapeutic choice for patients, and has a good application prospect. The medicine is especially suitable for patients with poor response to traditional medicines. Therefore, the sennoside B has a remarkable clinical application value and a wide market prospect, and a new choice is provided for the treatment of related diseases.

Description

Technical Field

[0001] The present invention relates to the technical field of medicines, and in particular to application of sennoside B in preparing a medicine for preventing and treating renal fibrosis caused by urinary tract obstruction. Background Art

[0002] Renal fibrosis is a typical pathological change in chronic kidney disease (CKD), and epithelial-mesenchymal transition (EMT) is the main stage. Tubulointerstitial fibrosis is characterized by tubular atrophy and extracellular matrix (ECM) accumulation in renal tissue. Since the expression level of TGF-β1 (transforming growth factor, β1) in cells is closely related to the production of EMT and ECM, it is considered to be a strong mediator of renal fibrosis and CKD and a potential key driver of renal fibrosis. Mitochondrial autophagy is a selective autophagy that can selectively degrade damaged or dysfunctional mitochondria. Mitochondrial autophagy reduces renal fibrosis by reducing the production of reactive oxygen species (ROS). Therefore, inhibiting the occurrence of EMT and regulating mitochondrial autophagy are of great significance for further and better treatment of renal injury. Pathological changes in CKD renal fibrosis is very common. When the ureter is obstructed, urine retention compresses the renal tubules, causing progressive necrosis of renal tubular epithelial cells, interstitial acute and chronic inflammatory cell infiltration, and necrotic renal tubular tissue is gradually replaced by fibrous scars, eventually forming progressive renal fibrosis. Because fibrosis is considered to be irreversible, inhibiting the progress of fibrosis is considered to be a potential strategy to prevent the development of CKD. At present, renal fibrosis becomes a medical problem to be solved urgently, and the present invention proposes a new solution for this reason. Therefore, it is of great clinical significance to alleviate or delay the renal interstitial fibrosis process in the CKD process and find new drugs for preventing and treating renal fibrosis. Summary of the invention

[0003] Sennoside B (SB) is a naturally occurring compound belonging to the class of anthrone glycosides. It is commonly found in traditional Chinese medicines such as senna leaves and rhubarb roots. It is widely used as a stimulant laxative, and its safety and effectiveness have been proven. A large number of experimental studies have shown that it has exhibited a series of pharmacological properties, such as anti-inflammatory and anti-tumor drug activities. In addition, Sennoside B can inhibit PDGF-stimulated cell proliferation by binding to PDGF-BB and its receptors and downregulating the PDGFR-β signaling pathway. In view of this, the present invention proposes the use of sennoside B in the preparation of drugs for preventing and treating renal fibrosis caused by urinary tract obstruction, aiming to provide a new prevention and treatment strategy to improve the therapeutic effect of renal fibrosis.

[0004] The present invention provides an application of sennoside B or a preparation thereof, wherein the application includes any one of the following: A1) Application of sennoside B in the preparation of drugs for preventing and treating renal fibrosis caused by urinary tract obstruction; A2) Use of sennoside B in a drug for inhibiting the epithelial-mesenchymal transition process induced by TGF-β1 in human renal tubular epithelial cells HK-2; A3) Application of sennoside B in increasing the expression of E-cadherin protein in human renal tubular epithelial cells HK-2; A4) Use of sennoside B in the preparation of a drug for increasing the expression of E-cadherin protein in human renal tubular epithelial cells HK-2; A5) Application of sennoside B in reducing the expression of N-cadherin protein in human renal tubular epithelial cells HK-2; A6) Use of sennoside B in the preparation of a drug for reducing the expression of N-cadherin protein in human renal tubular epithelial cells HK-2; A7) Application of sennoside B in reducing the expression of Vimentin protein in human renal tubular epithelial cells; A8) Use of sennoside B in the preparation of a drug for reducing the expression of Vimentin protein in human renal tubular epithelial cells; A9) Use of sennoside B in the preparation of a medicament for preventing and / or treating diseases associated with renal fibrosis.

[0005] A10) Application of sennoside B in reducing Fn protein expression in human glomerular mesangial cells (HRMC); A11) Use of sennoside B in the preparation of a drug for reducing the expression of Fn protein in human glomerular mesangial cells (HRMC); A12) Application of sennoside B in reducing COL protein expression in human glomerular mesangial cells (HRMC); A13) Use of sennoside B in the preparation of a drug for reducing the expression of COL protein in human glomerular mesangial cells (HRMC); Preferably, the disease associated with renal fibrosis is chronic kidney disease or nephrosclerosis caused by urinary tract obstruction.

[0006] The invention also provides a medicine for treating kidney disease, wherein the active ingredient of the medicine is sennoside B.

[0007] Preferably, the renal disease is renal fibrosis, chronic kidney disease or nephrosclerosis caused by urinary tract obstruction.

[0008] Compared with the prior art, the present invention has the following beneficial effects: 1. Provide a safer and more effective method for treating renal fibrosis, by utilizing the natural anti-inflammatory properties of sennoside B, reducing the side effects of drugs and improving the safety of treatment; 2. The sennoside B of the present invention can be used as a supplement or substitute for existing treatment methods, providing patients with new treatment options, especially for patients who do not respond well to traditional drugs; 3. The preparation process of sennoside B of the present invention is relatively mature, has high extraction efficiency and purity, is easy to produce and apply on a large scale, helps to reduce treatment costs, and makes it affordable for more patients; 4. The experimental results of the present invention show that sennoside B has a significant effect in reversing oxidative stress and improving mitochondrial function, which provides a new idea and method for developing new drugs for treating renal fibrosis.

[0009] 5. The experiments of the present invention also confirmed that sennoside B can significantly reduce the occurrence of mitochondrial autophagy. This regulatory mechanism helps to protect cells from damage caused by excessive autophagy, while maintaining the normal function of mitochondria and alleviating the occurrence of renal fibrosis, which provides a new therapeutic target for the treatment of renal fibrosis.

[0010] In summary, the present invention provides a new treatment strategy for preventing and treating renal fibrosis through the multiple pharmacological activities of sennoside B, has significant clinical application value and broad market prospects, and provides a new option for the treatment of related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Various other advantages and benefits will become apparent to those of ordinary skill in the art by reading the detailed description of the preferred embodiments below. The accompanying drawings are only for the purpose of illustrating the preferred embodiments and are not to be considered as limiting the present invention. Moreover, the same reference symbols are used throughout the accompanying drawings to represent the same components. In the accompanying drawings: Figure 1 This is the effect of sennoside B in Example 1 of the present invention on renal fibrosis in UUO rats.

[0012] Figure 2 This is the effect of sennoside B on HK-2 cell viability in Example 2 of the present invention.

[0013] Figure 3 This is the effect of Sennoside B in Example 3 of the present invention on the expression of E-cadherin, N-cadherin and Vimentin in HK-2 cells induced by TGF-β1.

[0014] Figure 4 This is the effect of sennoside B in Example 4 of the present invention on the opening degree of mitochondrial permeability transition pore of HK-2 cells induced by TGF-β1.

[0015] Figure 5 This is the effect of sennoside B in Example 5 of the present invention on mitochondrial membrane potential and mitochondrial reactive oxygen species in HK-2 cells induced by TGF-β1.

[0016] Figure 6 This is the effect of sennoside B in Example 6 of the present invention on TGF-β1-induced mitochondrial autophagy in HK-2 cells.

[0017] Figure 7 This is the effect of Sennoside B on TGF-β1-induced fibronectin and collagen in HRMC cells in Example 7 of the present invention. DETAILED DESCRIPTION

[0018] Exemplary embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. Although exemplary embodiments of the present disclosure are shown in the accompanying drawings, it should be understood that the present disclosure can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided in order to enable a more thorough understanding of the present disclosure and to fully convey the scope of the present disclosure to those skilled in the art. It should be noted that, in the absence of conflict, the embodiments of the present invention and the features described in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and in combination with the embodiments.

[0019] Example 1 Sennoside B improves renal fibrosis in UUO rats 1. Experimental Materials SD rats were provided by Sibeifu (Beijing) Biotechnology Co., Ltd. (Animal Certificate No.: NO.110324231106182758); Sennoside B (Cat. No.: T2734) was purchased from Shanghai Taoshu Biotechnology Co., Ltd.

[0020] 2. Experimental Methods Thirty-six SPF male SD rats weighing 150±10 g were selected and placed in a standard SPF environment. Standard feed was provided and rats were allowed to drink water freely. After one week of adaptive feeding with normal feed, the animals were randomly divided into six groups: normal control group, UUO model group, UUO model group + SB (2 mg / kg, 10 mg / kg, 50 mg / kg) group and UUO model group + pirfenidone group, 6 rats in each group. The mice were fasted the day before modeling. After fasting, CON, UUO group, UUO+SB group and UUO+pirfenidone group were modeled. When the rats were anesthetized, atropine sulfate was injected subcutaneously 15 minutes in advance, followed by intramuscular injection of Zotai 50 for anesthesia. The rats were fixed on the operating board in a lateral position, and a longitudinal incision was cut along the kidney position to expose the kidney. The normal group was not treated, while the model group and the drug-treated group underwent ureteral ligation. After the operation, the inner and outer skins were quickly sutured and disinfected. After the rats woke up, they were fed routinely and their condition was observed. After the model was successfully established, medication was started. The feeding methods of each group were as follows: Control group (sham operation group, also called normal group or Con group): provided with normal water and feed daily; UUO group (urinary tract obstruction renal fibrosis model group): normal water and feed were provided daily; UUO+SB group (Sennaside B group): Starting from the second day after surgery, daily intragastric administration of sennoside B was started, with the doses of 2 mg / kg, 10 mg / kg, and 50 mg / kg, respectively, and normal water and feed were provided daily for 14 days; UUO+PFD group (pirfenidone group): Pirfenidone was administered daily by gavage starting from the second day after surgery at a dose of 500 mg / kg, and normal water and feed were provided daily.

[0021] During the modeling and the above treatments, the rats were weighed weekly. On the 14th day of drug administration, the rats were anesthetized. The ultrasound workstation MyLab™X7, abdominal probe L 4-15, and the working frequency range were 4-15 MHz. All rats were in the supine position. The grayscale and depth were adjusted to the optimal state to observe the kidney morphology, size, cortical thickness, parenchymal echo and blood flow. After the B-ultrasound, the rats in anesthesia were killed and the renal cortical tissue was collected. The collected renal cortical tissue was fixed in 4% paraformaldehyde, dehydrated, embedded and sliced ​​to prepare paraffin sections. The sections were stained with hematoxylin-eosin staining (HE), Masson staining and glycogen (PeriodicAcid-Schiff stain, PAS) staining.

[0022] 3. Experimental Results Under ultrasonic testing ( Figure 1 A), it can be seen that compared with normal rats, the kidney morphology of UUO rats has changed, and the kidney volume has increased significantly. This is due to the obstruction of the ureter of the kidney on the surgical side, which hinders urine excretion, leading to increased pressure in the renal pelvis, dilation of the renal pelvis and calyces, and then hydronephrosis, which eventually leads to weight gain. Under color Doppler mode detection, red usually indicates blood flow toward the probe, and blue indicates blood flow away from the probe. Compared with normal rats, the renal blood flow distribution of UUO rats is less, and there may be vascular stenosis or occlusion after urine retention. Sennoside B and pirfenidone can improve the increase in renal pelvic pressure, dilation of the renal pelvis and calyces, and partially restore the renal blood flow of UUO rats, and the improvement effect of sennoside B is better than that of pirfenidone.

[0023] HE, PAS and Masson staining were used to evaluate the effects of sennoside B on renal histological changes and mesangial matrix deposition. HE staining showed that compared with normal rats, UUO rats had hydronephrosis, progressive dilatation of the renal pelvis and thinning of the renal cortex. Sennoside B and pirfenidone could improve tubular and glomerular damage, and the improvement effect of sennoside B was better than that of pirfenidone ( Figure 1 B). Masson staining showed increased glomerular collagen deposition in the kidneys of UUO rats ( Figure 1B), sennoside B and pirfenidone can reduce glomerular collagen deposition in UUO rats, and the effect of sennoside B is better than that of pirfenidone ( Figure 1 B). PAS staining showed that the PAS-positive mesangial matrix in the glomeruli of UUO rats increased compared with that in normal rats ( Figure 1 B), sennoside B and pirfenidone can reduce the PAS-positive mesangial matrix in the glomeruli of UUO rats, and the effect of sennoside B is better than that of pirfenidone ( Figure 1 B). It can be seen that sennoside B has the potential to improve renal fibrosis in UUO rats, providing a theoretical basis and experimental evidence for the development of new anti-fibrotic drugs.

[0024] Example 2 Effect of Sennoside B on HK2 Cell Viability 1. Experimental Materials Including human renal tubular epithelial cells HK-2 (provided by Shanghai Meiwan Biotechnology Co., Ltd.), human glomerular mesangial cells HRMC (provided by Shanghai Meiwan Biotechnology Co., Ltd.) and DMEM / F-12 (1:1) culture medium (purchased from Thermo Fisher Scientific).

[0025] 2. Experimental Methods To determine the appropriate drug concentration, CCK8 cell proliferation and toxicity assay kit was used to assess cell viability. Briefly, 1 × 10 4 HK-2 cells were seeded in 96-well plates and cultured for 24 h. After fully adhering to the wall, different doses of sennoside B (0, 0.5, 2.5, 5, 10, 20, 40, 80, 160 μM) were added to the culture medium and cultured for 48 h. After washing with PBS, basal culture medium containing CCK8 solution was added and incubated in the dark for 2 h. The absorbance was measured at 450 nm using an ELISA reader. In order to observe the cell morphology more intuitively, 8×10 cells were added to each well. 5 HK-2 cells were seeded in a 6-well plate and cultured for 24 h. After fully adhering to the wall, different doses of sennoside B (0, 0.5, 2.5, 5, 10, 20, 40, 80, 160 μM) were added to the culture medium and cultured for 48 h. After washing twice with PBS, the six-well plate was placed on the stage of a microscope and light microscopy images of the cells were taken.

[0026] 3. Experimental Results From light microscopy images of cells and CCK8 ( Figure 2 ) The results show that sennoside B has little effect on the viability of human renal tubular epithelial cells HK-2, and the cell morphology and viability remain basically normal.

[0027] Example 3 Sennoside B improves the expression of E-cadherin, N-cadherin and Vimentin proteins in human renal tubular epithelial cells HK-2 induced by TGF-β1 1. Experimental Materials The experimental materials followed the same materials as in Example 2.

[0028] 2. Experimental Methods (1) Cell culture and grouping: Place a sterile cover glass after burning in a six-well plate. After the cover glass cools down, inoculate HK-2 cells in the six-well plate at an appropriate density. When the cell density reaches 50%, replace the serum-free DMEM / F12 medium and place in the treatment for 12 h. Then divide the cells into six groups for different treatments. One group served as a blank control group with normal culture medium, another group was replaced with 10% FBS culture medium containing 10 ng / mL TGF-β1 as a model group, and the remaining three groups were added with different concentrations of sennoside B (2.5 μM, 10 μM, 40 μM) on the basis of the model group. The last group was added with pirfenidone (0.5 mg / mL) on the basis of the model group and cultured for 24 hours before subsequent experiments.

[0029] (2) Immunofluorescence detection: The expression of E-cadherin, N-cadherin, and Vimentin proteins in HK-2 cells was detected by immunofluorescence (IF). The specific steps of cell immunofluorescence detection are as follows: First, the collected cell samples were washed twice with PBS, then fixed with 4% paraformaldehyde at room temperature for 15 minutes, and then permeabilized with 0.1% Triton X-100 in PBS for 10 minutes; then, the cells were blocked with 10% goat serum for 30 minutes, and then incubated with the primary antibody containing 10% goat serum at 4°C overnight; then, they were washed three times with PBS, and then incubated with DyLight 488-conjugated AffiniPure goat anti-mouse IgG (H+L) or CY3-conjugated AffiniPure goat anti-rabbit IgG (H+L) in a dark box for 1.5 hours. The cell nuclei were stained with DAPI staining reagent for 30 minutes. Finally, the coverslip with cells was placed on a slide with anti-fluorescence quenching mounting medium, and a circle of nail polish was applied around the edge of the coverslip to fix the coverslip. After the coverslip was mounted, images were acquired using a single-photon confocal microscope.

[0030] 3. Experimental Results E-cadherin, as an epithelial cell-cell adhesion protein, had a weak fluorescence intensity in the control group. After TGF-β1 treatment, its fluorescence intensity increased, indicating increased cell-cell adhesion and aggravated fibrosis. After intervention with sennoside B, the fluorescence intensity showed a decreasing trend ( Figure 3 ). N-cadherin is a calcium-dependent transmembrane glycoprotein that is mainly expressed in mesenchymal cells. The fluorescence intensity was higher in the control group (Con group). After treatment with TGF-β1, the fluorescence intensity was significantly weakened. After intervention with sennoside B, the fluorescence intensity was enhanced ( Figure 3 ). Vimentin, as an intermediate filament protein, had a higher fluorescence intensity in the control group (Con group), and was significantly expressed in the cell-cell contact area, showing a concentrated specific fluorescence localization. After treatment with TGF-β1, the fluorescence intensity was significantly weakened. After intervention with sennoside B, the fluorescence intensity was enhanced ( Figure 3 ).

[0031] In summary, the effect of sennoside B on HK-2 cell fibrosis is manifested by regulating the expression of E-Cadherin, N-Cadherin and Vimentin, inhibiting the abnormal adhesion and migration ability between cells, and enhancing the structural stability of cells, thereby reducing the degree of fibrosis, and its effect is better than PFD.

[0032] Example 4 Sennoside B reduces the opening of the mitochondrial permeability transition pore in HK-2 cells 1. Experimental Materials The experimental materials were the same as those in Example 2 and the mitochondrial permeability transition pore (MPTP) detection kit was used.

[0033] 2. Experimental Methods The experimental method is as follows: (1) Cell culture and grouping followed the same procedures as in Example 3.

[0034] (2) Mitochondrial permeability transition pore detection: The degree of opening of the mitochondrial permeability transition pore was measured using the MPTP detection kit. The fluorescence quenching solution was a mixture of Calcein AM staining solution and CoCl2. The mixed working solution was applied to the cells for 60 minutes at 37°C, followed by incubation with fresh culture medium for 30 minutes, washing with PBS, and staining with Hoechst 33342 for 45 minutes. Images were acquired using a single-photon confocal microscope.

[0035] 3. Experimental Results The results showed that in HK-2 cells induced by TGF-β1, an increase in the openness of MPTP was observed, indicating impaired mitochondrial function. In HK-2 cells treated with sennoside B, the openness of MPTP was reduced ( Figure 4 ). The experimental results revealed that sennoside B can improve mitochondrial function by alleviating the openness of MPTP in HK2 cells.

[0036] Example 5 Sennoside B increases HK-2 cell mitochondrial membrane potential and reduces mitochondrial superoxide levels 1. Experimental Materials The experimental materials were the same as those in Example 2, including the mitochondrial membrane potential detection kit (JC-1) and the mitochondrial superoxide red fluorescent probe (Mito SOX).

[0037] 2. Experimental Methods The mitochondrial membrane potential (MMP) level was measured using a mitochondrial membrane potential assay kit containing JC-1. The probe was diluted to 1× concentration with JC-1 buffer, and the cells were incubated at 37°C for 90 minutes. The nuclei were counterstained with Hoechst 33342 staining solution at room temperature for 60 minutes, and images were acquired using a single-photon confocal microscope. Mitochondrial superoxide red fluorescence (MitoSOX Red) probe was used to detect mitochondrial superoxide. The probe was diluted to 5 µM with Hank's balanced salt solution (HBSS), and the cells were incubated at 37°C for 30 minutes in the dark. After washing twice with PBS, the nuclei were counterstained with Hoechst 33342 solution diluted in HBSS for 45 minutes, washed four times with PBS for 5 minutes each, and then images were acquired using a single-photon confocal microscope.

[0038] 3. Experimental Results The experimental results showed that in HK-2 cells induced by TGF-β1, a decrease in MMP was observed, indicating impaired mitochondrial function. However, after treatment with sennoside B, the decrease in MMP was significantly reversed ( Figure 5 ). At the same time, TGF-β1 induced a large amount of ROS in the mitochondria of HK-2 cells, and the ROS level in the mitochondria was significantly reduced after treatment with sennoside B. Based on the above results, sennoside B can improve the decrease of MMP and the increase of mitochondrial ROS level in HK-2 cells induced by TGF-β1, thereby improving mitochondrial function, and its effect is better than PFD.

[0039] Example 6 Sennoside B significantly reduces the level of mitochondrial autophagy in HK-2 cells induced by TGF-β1 1. Experimental Materials The experimental materials are the same as those in Example 2. Mitochondrial autophagy fluorescent probe (Mtphagy Dye), lysosomal fluorescent dye (Lyso Dye) and MitoTracker® Deep Red FM mitochondrial deep red fluorescent probe.

[0040] 2. Experimental Methods The cell culture and grouping followed the same procedure as in Example 3. HK-2 cells were seeded in glass-bottomed dishes at an appropriate density and cultured overnight. The culture medium was removed and the cells were washed twice with serum-free medium. Working solution 1 was prepared at a ratio of Mtphagy Dye storage solution: serum-free medium = 1:1000. 1 mL of working solution 1 was added to each glass-bottomed dish, and each group of cells was incubated at 37°C for 30 minutes. After incubation, the cells were washed twice with serum-free medium. Modeling drugs or therapeutic drugs of corresponding concentrations were added to the glass-bottomed dishes according to the grouping, and incubated at 37°C for 48 hours. After the culture was completed, the cells were washed twice with serum-free medium. Working solution 2 was prepared at a ratio of Lyso Dye storage solution: serum-free medium = 1:1000. 1 mL of working solution 2 was added to each glass-bottomed dish, and each group of cells was incubated at 37°C for 30 minutes. After the culture was completed, the cells were washed twice with serum-free medium. Working solution 3 was prepared at a ratio of MitoTracker® Deep Red FM storage solution: serum-free medium = 1:4000. 1 mL of working solution 3 was added to each glass-bottomed dish, and each group of cells was incubated at 37°C for 45 minutes. After the incubation, the cells were washed twice with serum-free medium. Finally, the cell nuclei were counterstained with Hoechst 33342 solution diluted in HBSS for 45 minutes, washed three times with serum-free medium, and then imaged using a single-photon confocal microscope.

[0041] 3. Experimental Results The experimental results show that under normal circumstances, the level of mitochondrial autophagy is relatively low, because the mitochondria in the cells function normally and there is no need to frequently remove damaged mitochondria ( Figure 6 When cells were stimulated by TGF-β1, mitochondrial autophagy increased significantly, indicating that TGF-β1 induces mitochondrial damage ( Figure 6 ). When sennoside B is added, the level of mitochondrial autophagy is reduced, inhibiting the signal of mitochondrial damage, thereby reducing the occurrence of mitochondrial autophagy ( Figure 6 This regulatory mechanism helps protect cells from damage caused by excessive autophagy, while maintaining the normal function of mitochondria and alleviating the occurrence of renal fibrosis, and its effect is better than that of PFD.

[0042] Example 7 Sennoside B significantly reduces the expression of fibronectin FN and collagen COL in HRMC cells 1. Experimental Materials The experimental materials followed the same materials as in Example 2.

[0043] 2. Experimental Methods The cell culture and grouping followed the same procedures as in Example 3. The immunofluorescence detection followed the same procedures as in Example 3.

[0044] 3. Experimental Results FN and COL both play a key role in renal fibrosis, and their overexpression promotes the progression of renal interstitial fibrosis. The experimental results showed that the content of FN and COL proteins in HRMC cells induced by TGF-β1 increased significantly, and sennoside B played a positive role in improving renal interstitial fibrosis by reducing the expression of FN and COL proteins, and its effect was better than PFD ( Figure 7 ).

[0045] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in the relevant field should understand that the specific implementation methods of the present invention can still be modified or replaced by equivalents. Any modification or equivalent replacement that does not depart from the spirit and scope of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. An application of sennoside B or its preparation, characterized in that: The application includes any of the following: A1) Application of sennoside B in the preparation of drugs for preventing and treating renal fibrosis caused by urinary tract obstruction; A2) Use of sennoside B in a drug for inhibiting the process of epithelial-mesenchymal transition induced by TGF-β1 in human renal tubular epithelial cells; A3) Application of sennoside B in increasing the expression of E-cadherin protein in human renal tubular epithelial cells; A4) Use of sennoside B in the preparation of a drug for increasing the expression of E-cadherin protein in human renal tubular epithelial cells; A5) Application of sennoside B in reducing the expression of N-cadherin protein in human renal tubular epithelial cells; A6) Use of sennoside B in the preparation of a drug for reducing the expression of N-cadherin protein in human renal tubular epithelial cells; A7) Application of sennoside B in reducing Vimentin protein expression; A8) Use of sennoside B in the preparation of a drug for reducing the expression of Vimentin protein; A9) Use of sennoside B in the preparation of a medicament for preventing and / or treating diseases associated with renal fibrosis.

2. An application of sennoside B or its preparation, characterized in that: The application also includes any of the following: B1) Application of sennoside B in reducing the expression of Fn protein in human glomerular mesangial cells; B2) Use of sennoside B in the preparation of drugs for reducing Fn protein expression; B3) Application of sennoside B in reducing COL protein expression in human glomerular mesangial cells; B4) Use of sennoside B in the preparation of a drug for reducing COL protein expression; 3. The use according to claims 1 and 2, characterized in that , the renal fibrosis-related improvements include increased intra-renal pelvic pressure, dilation of the renal pelvis and calyces, and restoration of renal blood flow.

4. The use according to claims 1 and 2, characterized in that The disease associated with renal fibrosis is chronic kidney disease or nephrosclerosis caused by urinary tract obstruction.

5. A drug for treating kidney disease, characterized in that: The active ingredient of the drug is sennoside B.

6. The drug according to claim 5, characterized in that The renal disease is renal fibrosis, chronic kidney disease or nephrosclerosis caused by urinary tract obstruction.

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