基于Raf/MEK/ERK通路探讨白芍总苷对肾结石大鼠肾纤维化的影响OA

[Objective]To discuss the effect of total glucosides of paeony(TGP)on renal fibrosis in rats with kidney stones(KS)based on the rapidly accelerated fibrosarcoma(Raf)/mitogen-activated extracellular signal-regulated kinase(MEK)/extracellular regulated protein kinase(ERK)pathway.[Methods]The successfully modeled KS rats were separated into the KS group,the low-dose TGP(L-TGP)group,the high-dose TGP(H-TGP)group(gavage of 100 and 200 mg/kg TGP),and the TGP+2-[(2-phenylethyl)thio]nicotinic acid(ML-099)group(gavage of 200 mg/kg TGP+intraperitoneal injection of 11.7 mg/kg ML-099),with 18 rats per group.The Ctrl group and the KS group were administered the same amount of normal saline by gavage and intraperitoneal injection once a day for 28 consecutive days.The renal functions of rats in each group were detected.The levels of interleukin(IL)-6 and IL-1β in the serum of KS rats were detected by ELISA kits.Von-Kossa staining was used to observe pathological calcium deposition in renal tissue.Renal histopathology was measured by HE and Masson staining.The levels of transforming growth factor β1(TGF-β1)and α-smooth muscle actin(α-SMA)in renal tissue were detected by immunohistochemistry.The expression of Raf/MEK/ERK pathway proteins in renal tissue was measured by Western blot.[Results]The renal tissue structure in the Ctrl group was intact,and no obvious collagen fibers or CaOx crystals were observed.In the KS group,the renal tissue structure was mixed,the renal tubules were dilated or deformed,obvious inflammatory cell infiltration,collagen fiber deposition and CaOx crystals could be observed,and the CVF and CaOx crystal scores increased.The pathological damage of renal tissues in the L-TGP and H-TGP groups improved successively,the collagen fiber deposition and CaOx crystals decreased successively,and the CVF and CaOx crystal scores decreased.In the TGP+ML-099 group,renal tissue injury was further aggravated,collagen fiber deposition and CaOx crystals were further increased,and CVF and CaOx crystal scores were elevated.The BUN,Scr,24 h urine protein,IL-6,IL-1β,TGF-β1,α-SMA,p-Raf/Raf,p-MEK/MEK,and p-ERK/ERK in the KS group were higher than those in the Ctrl group(P<0.05).The BUN,Scr,24 h urine protein,IL-6,IL-1β,TGF-β1,α-SMA,p-Raf/Raf,p-MEK/MEK and p-ERK/ERK in the L-TGP and H-TGP groups were lower than those in the KS group(P<0.05).The BUN,Scr,24 h urine protein,IL-6,IL-1β,TGF-β1,α-SMA,p-Raf/Raf,p-MEK/MEK,and p-ERK/ERK in the TGP+ML-099 group were higher than those in the H-TGP group(P<0.05).[Conclusion]TGP may suppress the inflammatory response,and improve renal fibrosis in KS rats by inhibiting the Raf/MEK/ERK pathway.