摘要
目的探讨芪蓟多糖通过NF-κB信号通路对大鼠肾小球系膜细胞的影响。方法体外培养大鼠肾小球系膜细胞,用血管紧张素Ⅱ(AngⅡ)刺激系膜细胞增生,分别给予氯沙坦和芪蓟多糖,用酶联免疫吸附法(ELISA)检测肾小球系膜细胞上清液转化生长因子β活化激酶1(TAK1)、肿瘤坏死因子受体相关因子6(TRAF6)、核因子(NF-κB)和纤维粘连蛋白(FN)表达;实时定量PCR法检测TAK1、NF-κB的表达。结果 ELISA法对OD值的测定,与正常组比较,模型组、芪蓟肾康方多糖高剂量组和低剂量组、氯沙坦组大鼠肾小球系膜细胞中TAK1、TRAF6、NF-κB和FN表达均升高且有明显差异(P<0.05,P<0.01);与模型组比较,芪蓟肾康方多糖高剂量组和低剂量组、氯沙坦组可降低NF-κB信号转导中各靶点TAK1、TRAF6、NF-κB及FN的表达,且有明显差异(P<0.01)。实时定量PCR法检测,与正常组比较,模型组及芪蓟肾康方多糖高剂量组和低剂量组大鼠肾小球系膜细胞中TAK1、NF-κB表达升高,且有明显差异(P<0.05);与模型组比较,芪蓟肾康方多糖高剂量组和低剂量组大鼠肾小球系膜细胞中TAK1、NF-κB明显降低(P<0.05)。在NF-κB表达中,芪蓟肾康方多糖高剂量组和低剂量组组间比较无明显差异(P>0.05)。结论芪蓟多糖可能通过下调肾小球系膜细胞NF-κB mRNA及TAK1、TRAF6、NF-κB和FN蛋白表达的水平,抑制核因子信号转导通路的兴奋性,缓解NF-κB的释放,减少FN的过度表达,从而减轻细胞外基质(ECM)的分泌,降低肾小球的损伤,减缓肾小球的硬化。
Objective To explore the impacts of Qiji polysaccharose on glomerular mesangial cells via NF - κB signal pathway. Methods The glomerular mesangial cells were cultured in vitro. Ang II was used to stimulate glomerular mesangial cell proliferation. Losartan and Qiji polysaccharose were applied sepa-rately. ELISA was applied to determine the expressions of TAK1,TRAF6,NF - κB and FN in the supernate of glomerular mesangial cells. The real - time quantitative PCR was used to determine the expressions of TAK1 and NF - κB. Results In ELISA determination of OD value,compared with the normal group,the expres-sions of TAK1,TRAF6,NF - κB and FN in glomerular mesangial cells were all increased in the model group, high - dose group and low - dose group of Qiji polysaccharose,as well as Losartan group,indicating the ap-parent differences（P ﹤ 0. 05,P ﹤ 0. 01）. Compared with the model group,the expressions of TAK1,TRAF6, NF - κB and FN at each target point of NF - κB transduction were reduced in the high - dose group,low -dose group and Losartan group（P ﹤ 0. 01）. In real - time quantitative PCR determination,compared with the normal group,the expressions of TAK1 and NF - κB in glomerular mesangial cells were increased in the mod-el group,high - dose group and low - dose group of Qiji polysaccharose,indicating the obvious difference（P﹤ 0. 05）. Compared with the model group,the expressions of TAK1 and NF - κB in glomerular mesangial cells were reduced apparently in the model group,high - dose group and low - dose group of Qiji polysaccharose.Concerning to NF - κB expression,the difference was not significant between the high - dose group and low -dose group of Qiji polysaccharose（P ﹥ 0. 05）. Conclusion Qiji polysaccharose inhibits the excitability of nuclear factor signal transduction pathway,relieves NF - κB release,reduces FN over - expression via down- regulating the expressions of TAK1,NF - κB mRNA and the protein expressions of TAK1,TRAF6,NF -κB and FN in glomerular mesangial cells so as to alleviate ECM secretion,reduce glomerular damage and re-tard glomerular sclerosis.
出处
《世界中西医结合杂志》
2014年第6期644-647,共4页
World Journal of Integrated Traditional and Western Medicine
基金
国家科技重大专项资助项目(No.2009ZX09103)
