不同剂量维拉帕米对内毒素血症大鼠肝细胞因子表达和NF-κB活化的影响

Verapamil modulates LPS-induced cytokines production via inhibiting NF-κB activation in the liver of rats

目的:观察不同剂量维拉帕米对内毒素(LPS)刺激的大鼠肝致炎/抗炎细胞因子的表达及对核因子κB(NF-κB)活化的调控作用。方法:56只SD大鼠随机分为七组:A组为等渗盐水对照组;B组为等渗盐水+LPS 10mg/kg;C、D、E、F组为不同剂量维拉帕米组,分别给予维拉帕米1、2.5、5和10 mg/kg+LPS 10 mg/kg;G组为维拉帕米对照组,维拉帕米10 mg/kg。取大鼠肝匀浆和血清,酶联免疫吸附法(ELISA)测定肝组织中炎性因子如肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6)、IL-10。电泳迁移率变动分析实验(EMSA)测定肝NF-κB的表达,同时检测血清转氨酶水平。结果:内毒素可诱导肝组织中TNF-α、IL-6和IL-10表达增加(P<0.01),血清谷丙转氨酶(ALT)和谷草转氨酶(AST)水平升高(P<0.01),同时诱导肝组织NF-κB的活化(P<0.01)。维拉帕米可不同程度地降低内毒素诱导的肝TNF-α、IL-6和NF-κB的生成,降低血清ALT和AST水平,增加肝组织IL-10的表达,并且与维垃帕米剂量相关。结论:不同剂量维拉帕米以剂量依赖方式调节内毒素诱导的大鼠肝致炎/抗炎因子的表达,同时抑制NF-κB的活化,改善内毒素诱导的急性肝损伤。

Objective：To investigate the effect of verapamil on the endotoxin-induced cytokine production and nuclear factor kappa B （NF-κB） activation in the liver. Methods：Fifty six adult male Sprague-Dawley rats were randomly divided into seven groups： group A： saline ; group B：endotoxin （10 mg/ kg, intraperitoneally, i. p. ） ; group C, D, E, F：endotoxin （10 mg/kg） after treatment with verapamil （1, 2.5, 5, 10 mg/kg i.p. respectively）, and group G：verapamil alone （10 mg/kg）. Tumor necrosis factor （TNF-α）, interleukin-6 （IL-6）, intedeukin-10 （IL-10） and NF-κB in the liver tissues and the serum levels of ALT and AST were investigated at 1 h afer LPS injection. Results ： Endotoxin stimulated production of TNF-α, IL-6 and IL-10, and activated NF-κB in the liver. Verapamil attenuated acute liver injury, down-regulated endotoxin-induced pro-inflammatory cytokine production, up-regulated ant-inflammatory cytokines production and inhibited NF-κB activation in the liver in a dose-dependent manner. Conclusion ： Verapamil can attenuate acute liver injury by down-regulating the production of TNF-α, IL- 6 and up-regulating IL-10 in the liver in a dose-dependent manner, possibly via inhibition of NF-κB.