刺五加注射液通过TLR4/NF-κB通路抑制肾缺血再灌注大鼠炎症反应的研究

Acanthopanax senticosus injection inhibits inflammation in rats with renal ischemia-reperfusion via TLR4/NF-κB pathway

目的探讨刺五加注射液(ASI)对肾缺血再灌注(RIR)大鼠炎症反应的影响及其机制。方法将96只健康SD大鼠随机分为假手术(Sham)组,模型(Model)组,ASI(30 mg/kg)组和ASI(30 mg/kg)+LPS(TLR4激活剂,5 mg/kg)组,每组24只,计算各组大鼠肾脏指数,检测血清肾功能指标。HE染色法行肾组织病理学检查并行病理评分;ELISA法检测血清和肾组织炎症因子水平;RT-PCR法、Western blot法分别检测肾组织Toll样受体4(TLR4)、核因子-κB p65(NF-κB p65)mRNA和蛋白表达。结果ASI显著降低RIR大鼠肾脏指数和血清尿素氮(BUN)、肌酐(SCr)水平(P<0.05);明显改善肾小球体积增大、肾小管管腔扩张、细胞空泡变性、炎性细胞浸润等病理学改变,降低病理评分(P<0.05);降低血清和肾组织TNF-α、IL-1β水平,提高IL-10水平(P<0.05);降低肾组织TLR4、NF-κB p65 mRNA和蛋白表达量(P<0.05)。LPS明显逆转ASI对TLR4、NF-κB p65 mRNA和蛋白表达的调控作用。结论ASI具有抑制RIR大鼠炎症反应、减轻肾组织损伤的作用,其机制可能与抑制TLR4/NF-κB通路有关。

Objective To investigate the effect of acanthopanax senticosus injection(ASI)on the inflammatory response in rats with renal ischemia reperfusion(RIR)and its mechanism.Methods 96 healthy SD rats were randomly divided into Sham group,Model group,ASI(30 mg/kg)group and ASI(30 mg/kg)+LPS(TLR4 activator,5 mg/kg)group,n=24.The renal index of rats was calculated.The level of renal function indexes in serum were detected.The renal histopathological was examined by HE staining method and the pathological damage was scored.The level of inflammatory factors in serum and renal tissue were detected by ELISA method.The mRNA and protein expressions of Toll-like receptor 4(TLR4),nuclear factor-κB p65(NF-κB p65)in renal tissue were detected by RT-PCR and Western blot method.Results ASI significantly decreased renal index and the level of blood urea nitrogen(BUN),creatinine(SCr)in serum of the rats with RIR(P<0.05);significantly improved the renal histopathological changes such as enlargement of glomerulus,dilation of renal tubule lumen,vacuolar degeneration of cells and inflammatory cell infiltration,decreased the pathological score(P<0.05);decreased the level of TNF-α,IL-1βin serum and renal tissue,increased the level of IL-10(P<0.05);decreased the mRNA and protein expression of TLR4,NF-κB p65(P<0.05).LPS significantly reversed the regulatory effect of ASI on the mRNA and protein expression of TLR4,NF-κB p65.Conclusion ASI can inhibit the inflammatory response of RIR rats and reduce the damage of renal tissue,which mechanism may be related to the inhibition of TLR4/NF-κB pathway.