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1.
Article in Chinese | WPRIM | ID: wpr-906515

ABSTRACT

Objective:To observe the effect of modified Shengjiangsan on renal fibrosis in rats with membranous nephropathy (MN) and to explore the mechanism of its complications of renal fibrosis. Method:Rats were injected with cationized bovine serum albumin(C-BSA)in the tail vein to establish a rat model of membranous nephropathy. The normal group,model group,modified Shengjiangsan group (27.3 g·kg<sup>-1</sup>)and benazepril group(10 mg·kg<sup>-1</sup>)were established in this study. Each group was given corresponding dosage of the drug once a day for 4 weeks of continuous intervention. After the administration,we observed the pathological changes of rat kidneys by the technology of Masson staining, silverhexylamine iodate (PASM) staining, transmission electron microscopy (TEM), immunofluorescence technology (IF) was used to detect immunoglobulin(Ig)G deposition in rat kidneys. The levels of interleukin-1<italic>β</italic> (IL-1<italic>β</italic>), interleukin-6 (IL-6), tumor necrosis factor-<italic>α</italic> (TNF-<italic>α</italic>) in rat serum were detected by enzyme-linked immunosorbent assay (ELISA) method. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and immunohistochemistry (IHC) were used to detect the mRNA and protein expression levels of monocyte chemotactic protein 1 (MCP-1), intercellular adhesion molecule 1 (ICAM-1), nuclear factor kappa B (NF-<italic>κ</italic>B), Toll-like receptor 4 (TLR4), plasminogen activator inhibitor 1 (PAI-1), transforming growth factor-<italic>β</italic><sub>1</sub> (TGF-<italic>β</italic><sub>1</sub>), <italic>α</italic>-smooth muscle actin (<italic>α</italic>-SMΑ) and type Ⅳ Collagen (Collagen Ⅳ) in rat kidney tissues. Result:Compared with normal group, the kidney tissue of the model group was obviously fibrotic, the serum levels of IL-1<italic>β</italic>, IL-6, and TNF-<italic>α</italic> were significantly increased(<italic>P</italic><0.05), and the expressions of MCP-1, ICAM-1, NF-<italic>κ</italic>B, TLR4, PAI-1, TGF-<italic>β</italic><sub>1</sub>, <italic>α</italic>-SMA and Collagen Ⅳ mRNA and protein in kidney tissue were significantly increased(<italic>P</italic><0.05). Compared with model group, modified Shengjiangsan and benazepril significantly improved renal fibrosis in rats, reduced the levels of IL-1<italic>β</italic>, IL-6, and TNF-<italic>α</italic> in the serum of MN rats(<italic>P</italic><0.05), down-regulated MCP-1, ICAM-1, NF-<italic>κ</italic>B, TLR4, PAI-1, TGF-<italic>β</italic><sub>1</sub>, <italic>α</italic>-SMA and Collagen Ⅳ mRNA and protein expression in kidney tissue(<italic>P</italic><0.05). Conclusion:Modified Shengjiangsan can reduce the release and expression of inflammatory factors by down-regulating the TLR4/NF-<italic>κ</italic>B signaling pathway, inhibit renal fibrosis, and reduce renal damage in MN rats.

2.
Article in Chinese | WPRIM | ID: wpr-906051

ABSTRACT

Objective:To investigate the intervention effect of modified Shengjiangsan on hypoxia-inducible factor-1<italic>α </italic>(HIF-1<italic>α</italic>)/nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) signaling pathway in membranous nephropathy (MN) rats and to explore its mechanism to reduce oxidative stress and apoptosis in renal tissues. Method:Cationized bovine serum albumin (C-BSA) was injected into the tail vein of rats to replicate the MN model. Rats were randomly divided into a model group, a modified Shengjiangsan group, and a benazepril group after modeling, and administered by gavage once a day accordingly. At the end of the 4<sup>th</sup> week, the 24-h urine total protein (UTP), urea nitrogen (BUN), and serum creatinine (SCr) levels of each group were detected. Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of superoxide dismutase (SOD), malondialdehyde (MDA), and reactive oxygen species (ROS) in renal tissues of rats. In situ end labeling(TUNEL) staining was used to detect the cell apoptosis rate. The mRNA and protein expression levels of HIF-1<italic>α</italic> and NOX4 were detected by real-time fluorescence-based quantitative polymerase chain reaction(Real-time PCR)and Western blot, respectively. The immunohistochemistry method was used to detect the protein expression levels of B-cell lymphomas -2 (Bcl-2), B-cell lymphomas xl (Bcl-xl), Bcl-2 associated X protein (Bax), Bcl-2 cell death regulator antibody (Bim). Result:Compared with the normal group, the model group showed increased UTP (<italic>P</italic><0.05), decreased SOD, elevated MDA and ROS (<italic>P</italic><0.05), up-regulated mRNA and protein expression of HIF-1<italic>α</italic> and NOX4 (<italic>P</italic><0.05), enhanced protein expression of Bax and Bim, declining protein expression of Bcl-xl and Bcl-2 (<italic>P</italic><0.05), and increased cell apoptosis in renal tissues. Compared with the model group, the modified Shengjiangsan group and the benazepril group displayed declining UTP (<italic>P</italic><0.05), up-regulated SOD, decreased MDA and ROS (<italic>P</italic><0.05), down-regulated mRNA and protein expression of HIF-1<italic>α</italic> and NOX4 (<italic>P</italic><0.05), diminished protein expression of Bax and Bim, elevated protein expression of Bcl-xl and Bcl-2 (<italic>P</italic><0.05), and reduced cell apoptosis in renal tissues (<italic>P</italic><0.05). Conclusion:The protective effect of modified Shengjiangsan on the kidney is presumedly achieved by reducing the oxidative stress and apoptosis in renal tissues of MN rats via inhibiting the HIF-1<italic>α</italic>/NOX4 signaling pathway.

3.
Article in Chinese | WPRIM | ID: wpr-905829

ABSTRACT

Objective:The purpose of this article was to observe the effect of modified Shengjiangsan on podocyte apoptosis in membranous nephropathy (MN) rats, to explore the molecular mechanism of its treatment of MN and to provide experimental basis for its clinical application. Method:The MN rat model was established by injection of cationic bovine serum albumin into the tail vein of rats. The successfully modeled rats were then randomly divided into model group (equal volume of normal saline), modified Shengjiangsan group (27.3 g·kg<sup>-1</sup>) and benazepril group (10 mg·kg<sup>-1</sup>), with corresponding drug dosage once a day for 4 weeks of continuous intervention. After drug administration, the 24-hour urine protein (UTP) was detected. Real time fluorescent quantitative polymerase chain reaction (Real-time PCR) and immunohistochemical (IHC) methods were used to detect Podocalyxin, Nephrin, Podocin, Synaptopodin mRNA and protein expression levels in rat kidney tissue. terninal-deoxynucleoitidyl transferase medsated nick and labeling (TUNEL) method was used to detect cell apoptosis rate in rat kidney tissue, and Western blot was used to detect Notch1, Hes1, B lymphoblastoma-2 (Bcl-2) associated X protein (Bax), and Bcl-2 protein expression levels in rat kidney tissue. Result:Compared with the normal group, UTP in the model group increased significantly, renal tissue cell apoptosis increased significantly, podocyte marker proteins podocalyxin, Nephrin, Podocin, Synaptopodin mRNA and protein expression levels decreased significantly, and Notch1, Hes1, Bax protein expression increased significantly, and Bcl-2 protein expression was significantly reduced(<italic>P</italic><0.05). Compared with the model group, UTP levels in MN rats were significantly reduced in modified Shengjiangsan and benazepril groups, with reduced rate of renal cell apoptosis, increased mRNA and protein expression levels of podocalyxin, Nephrin, Podocin, and Synaptopodin in renal tissue, decreased Notch1, Hes1, Bax protein expression, and increased Bcl-2 protein expression(<italic>P</italic><0.05). Conclusion:Modified Shengjiangsan can inhibit the Notch signaling pathway, reduce the apoptosis of rat kidney tissue podocytes, and reduce the kidney injury of MN rats.

4.
Article in Chinese | WPRIM | ID: wpr-872820

ABSTRACT

Objective:To explore the intervention effect of modified Shengjiangsan in rats with membranous nephropathy (MN) and its related mechanism. Method:Rats were injected with cationized bovine serum Albumin (C-BSA) in the tail vein to establish a rat model of membranous nephropathy. The normal group, model group, modified Shengjiangsan group (27.3 g·kg-1) and benazepril group (10 mg·kg-1) were established in this study. Each group was given corresponding dosage of the drug once a day for 4 weeks of continuous intervention. After the administration, the levels of 24-hour urine protein (UTP), total cholesterol (TC), triglyceride (TG), total protein (TP), Albumin (Alb), creatinine (SCr), urea nitrogen (BUN) level was detected. we observed the pathological changes of rat kidneys by the technology of Masson staining, silver hexylamine iodate (PASM) staining and transmission electron microscopy. immunofluorescence technology was used to detect immunoglobulin (Ig)G deposition in rat kidneys. Western blot was used to detect the expression levels of key proteins in phosphatidylinositol 3-kinase/proline protein kinase B/rapamycin target protein (PI3K/Akt/mTOR) signaling pathway and autophagy marker proteins LC3 and Beclin1. Result:Compared with normal group, the UTP, serum TC and TG levels were significantly increased, TP and Alb levels were significantly reduced in model group(P<0.05). We detected the kidney pathological changes include of glomerulus enlargement, basement membrane thickening,vacuolar degeneration, pheotropin deposition, glomerular capillary loop IgG diffuse deposition, electron dense deposits of varying sizes and podocytes under the epithelium extensive integration of foot processes, the expression of p-PI3K, p-Akt and p-mTOR protein was significantly increased (P<0.05). The expression of autophagy marker proteins LC3 and Beclin1 protein decreased significantly(P<0.05). Compared with model group, the UTP, serum TC and TG levels were decreased in the benazepril group and modified Shengjiangsan group, and the TP and Alb levels were increased (P<0.05), the histopathological changes of rat kidney were all reduced, the expression of p-PI3K, p-Akt and p-mTOR protein was significantly reduced(P<0.05), autophagy marker proteins LC3 and Beclin1 protein expression were significantly increased. Conclusion:Modified Shengjiangsan can reduce urinary protein, reduce kidney pathological damage and delay disease progression, which is related to its inhibition of PI3K/Akt/mTOR signaling pathway and activation of renal autophagy.

5.
Article in Chinese | WPRIM | ID: wpr-802239

ABSTRACT

Objective: To detect the effect of modified Shengjiangsan on the expression of reactive oxygen species(ROS) in mitochondria of renal foot cells of rats, in order to study the mechanism of modified Shengjiangsan. Method: After be fed for 7 days,the 60 SD male rats were randomly divided into four groups:blank control group, model group, positive medicine group and traditional Chinese medicine(TCM) treatment group. After establishment of the rat model of membranous nephropathy, model group, positive medicine group and TCM treatment group were treated differently. After 4 weeks, all of the rats were put to death, and the expressions of ROS, 24-hour urinary protein quantity,total cholesterol,triglyceride,total protein,albumin,urea nitrogen,creatinine were detected by Real-time fluorescence quantitative polymerase chain reaction Real-time PCR. Result: The expression of 24-hour urinary protein quantity,total cholesterol,triglyceride in positive medicine group and TCM treatment group were reduced,and the expressions of total protein,albumin in positive medicine group and TCM treatment group were reduced compared with those of model group (PPConclusion: Modified Shengjiangsan can effectively control the development of ROS in mitochondria of renal foot cells of rats, and repair the renal function of membranous nephropathy rats by recovering foot cells.

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