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  • Title: Zhen-wu-tang ameliorates membranous nephropathy rats through inhibiting NF-κB pathway and NLRP3 inflammasome.
    Author: Liu B, Lu R, Li H, Zhou Y, Zhang P, Bai L, Chen D, Chen J, Li J, Yu P, Wu J, Liang C, Song J, Liu X, Zhou J.
    Journal: Phytomedicine; 2019 Jun; 59():152913. PubMed ID: 30991182.
    Abstract:
    BACKGROUND: Zhen-wu-tang (ZWT), a traditional herbal formula, has been widely used for the treatment of kidney diseases in clinics, but the underlying molecular mechanisms have not been fully understood. PURPOSE: Inflammation mediated podocyte injury has been reported to constitute a crucial part in the pathogenesis of membranous nephropathy (MN). The current study was designed to evaluate the effect of ZWT on MN related to nuclear factor-κB (NF-κB) pathway and NLRP3 inflammasome. METHODS: The main components of ZWT were identified by 3D-ultra performance liquid chromatography (3D-UPLC) assay. A MN rat model induced by cationic-bovine serum albumin (C-BSA) and podocytes stimulated by TNF-α were used in this study. The 24 h urine protein, serum total cholesterol (TC) and triglyceride (TG), as well as kidney histology were measured to evaluate kidney damage. The expressions of IgG and complement 3 (C3), and the co-localization of NLRP3 and ASC were detected by immunofluorescence. The expressions of podocyte injury related protein desmin, podocin were measured by immunohistochemistry and western blot. Cell vitality of cultured podocytes was detected by MTT assay, as apoptosis assay was measured via flow cytometry. The protein expressions of p-p65, p-IκBα, NLRP3, Caspase-1, IL-1β were detected by western blot. RESULTS: Our results showed that ZWT significantly ameliorated kidney damage in MN model rats by decreasing the levels of 24 h urine protein, TC and TG. ZWT also improved renal histology and reduced the expressions of IgG and C3 in glomerulus. In addition, ZWT lessened the expressions of desmin, but increased podocin expression in vivo and vitro. ZWT protected cultured podocytes by maintaining cell vitality and inhibiting apoptosis. Moreover, we found that ZWT suppressed the expressions of NLRP3, Caspase-1, IL-1β and the co-localization of NLRP3 and ASC. Furthermore, the inhibition of NLRP3 inflammasome under ZWT treatment were accompanied by down-regulation of NF-κB pathway, as the p-p65 and p-IκBα protein expression were reduced. CONCLUSIONS: Our present study indicates that the inhibition of NF-κB pathway and NLRP3 inflammasome might be the potential mechanisms for the therapeutic effects of ZWT against MN.
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