护肾固精方下调核因子-κB活化对系膜细胞肿瘤坏死因子-α表达的影响

目的 :研究护肾固精方 (Hushengujingfang ,HSGJF)对脂多糖 (LSP)刺激大鼠肾小球系膜细胞(mesangialcell,MC)表达炎性细胞因子的影响及其作用机制。方法 :分别以酶联免疫吸附法 (ELISA)检测MCIL 1β、TNF α蛋白水平 ;电泳迁移率变动分析 (EMSA)检测MC核因子 κB(nuclearfactor κB ,NF κB)的活性 ;反转录多聚酶链反应技术 (RT PCR)检测MCTNF αmRNA表达。结果 :HSGJF能抑制LSP刺激肾小球MC分泌IL 1β、TNF α蛋白质增加的同时 ,亦能抑制LSP刺激肾小球MC中NF κB活化 ,在HSGJF作用下炎性细胞因子表达均呈不同程度减弱。结论 :HSGJF通过抑制MCNF κB的活性 ,下调炎性细胞因子的表达。此结果可作为进一步认识和评价该方对肾脏疾病防治作用的实验依据。

Inhibitory effect of HSGJF on LSP-induced expression of TNF-α in mesangial cells by down-regulated activation of nuclear factor-κB

Objective:To study the effects of hu shen gu jing fang(HSGJF) on LSP which stimulates culture rats glomerular mesangial cells(MSC) to produce proinflammatory cytokines and its machanisms. Methods:The protein synthesis of IL-1β and TNF-α in mesangial cells were measured by ELISA.The activation of NF-κB in mesangial cells was determined by electrophoresis mobility shift assay(EMSA).mRNA expression of proinflammatory cytokines (TNF-α) was examined by RT-PCR.Results: LSP induced activation of NF-κB and the expression of IL-1β and TNF-α in protein level in mesangial cells.HSGJF could inhibit the above effect of LSP.Conclusions: HSGJF may decrease the production of proinflammatory cytokines through the inhibition of NF-κB activation.This provides an theoretical and experimental evidence for evaluating the renal protective effect of HSGJF.