Renoprotective Effect of Sanjiao Qushi Prescription Against Cationic Bovine Serum Albumin Induced Membranous Nephropathy Mice Model and Its Influence on Nrf2/HO-1 Signaling Pathway / 中国实验方剂学杂志

ABSTRACT

Objective:To investigate the renoprotective effects that Sanjiao Qushi prescription ameliorates cationic bovine serum albumin （C-BSA） induced membranous nephropathy（MN） in mice model and its influence on nuclear factor erythroid-2-related factor-2（Nrf2）/heme oxygenase-1（HO-1） signaling pathway. Method:Sixty female BALB/c mice were randomly divided into the normal group（<italic>n</italic>=10） and the model group（<italic>n</italic>=50）. The mice in the model group received C-BSA injection via tail vein （6.5 mg·kg^-1）. The mice that were successfully modeled were randomized into the model group， the low dose Sanjiao Qushi prescription group（3.71 g·kg^-1）， the high dose Sanjiao Qushi prescription group（7.42 g·kg^-1） and benazepril hydrochloride group（1.3 mg·kg^-1）. And they were administered with the corresponding medicine by gavage once a day for four consecutive weeks. 24 hour-urine protein quantitation were performed before C-BSA injection and after C-BSA injection as well as the medicine gavage. When the treatment was finished， all of the mice were sacrificed and the biochemical indicators such as serum creatinine（SCr）， blood urea nitrogen（BUN）， triglyceride（TG）， total cholesterol（TC）， total protein（TP） and albumin（Alb） were measured. And the renal pathological morphology changes were observed by light microscope with hematoxylineosin（HE）， Masson and periodic acid-silver metheramine（PASM） staining. The deposition of immunoglobulin G（IgG） in the glomerulus was detected by fluorescence microscope. The expression of reactive oxygen species （ROS） of kidney was detected by fluorescence immunoassay. The protein expression levels of Nrf2 in cell nucleus and cytoplasm and the downstream protein factors HO-1 and NADH quinone acceptor oxidoreductase 1（NQO1） were detected by Western blot. Result:Compare to normal group， the levels of 24 hour-urine protein quantitation， TG and TC significantly increased in model group（<italic>P</italic><0.01）， while TP and Alb levels significantly decreased（<italic>P</italic><0.01）. The model group exhibited enlarged volume of glomerular， significantly thickened glomerular basement membrane（GBM）， fuchsinophilic protein deposition and spike formation through light microscope. Immunofluorescence staining for the model group exhibited granular deposition of IgG along the capillary wall. The expression of ROS in kidney significantly increased（<italic>P</italic><0.01）. The protein expression levels of Nrf2 in cell nucleus significantly increased（<italic>P</italic><0.01）， while Nrf2 in cytoplasm significantly decreased（<italic>P</italic><0.01）.The protein expression levels of HO-1 and NQO1 significantly increased（<italic>P</italic><0.01）. Compared to model group， the levels of 24 hour-urine protein quantitation， TG and TC significantly decreased in each treated group（<italic>P</italic><0.01）， TP and Alb levels significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The pathological damages alleviated obviously. The expression of ROS in kidney significantly decreased（<italic>P</italic><0.01）. The protein expression levels of Nrf2 in cell nucleus significantly decreased（<italic>P</italic><0.01）， while Nrf2 in cytoplasm significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The protein expression levels of HO-1 and NQO1 significantly increased（<italic>P</italic><0.01）. Conclusion:Sanjiao Qushi prescription worked on MN mice possibly by regulating related proteins in the Nrf2/HO-1 signaling pathway and relieving oxidative stress， thus decreasing 24 hour-urine protein and blood lipid， increasing serum protein， and alleviating the pathological damages to protect renal function and delay progress of the disease.