摘要
目的:以清热解毒中药金银花、连翘等为研究对象,重点观察其对TNF-α诱导的SV40 MES 13 NF-κB信号通路的影响,探讨清热解毒中药治疗肾脏病的有效机制。方法:通过MTT检测10味清热解毒中药对SV40 MES 13生长的影响,从中选出毒性小、作用浓度范围广的3种中药金银花、连翘、蒲公英作为研究对象分别作用于TNF-α刺激的SV40MES 13细胞,MTT检测其增殖、ELISA检测细胞浆p-IκBα、细胞核和细胞浆NF-κBp65及培养上清液中MCP-1、IL-6的表达。结果:(1)TNF-α能明显促进SV40 MES 13的增殖(P<0.05或P<0.01),药物处理组能抑制TNF-α刺激的SV40MES 13的增殖(P<0.01)。TNF-α与药物联合作用时抑制作用比单独药物处理更明显,呈浓度依赖性。(2)ELISA结果显示不同药物处理组细胞浆p-IκBα、细胞核NF-κBp65蛋白表达明显减低(P<0.01),细胞培养上清液中MCP-1、IL-6的含量明显降低(P<0.01),相同浓度的3种中药以金银花作用更明显。提示清热解毒药可以浓度依赖性地抑制SV40 MES 13的IκBα的磷酸化,减少细胞核NF-κB p65表达,阻止NF-κB核转位,从而抑制NF-κB活性,减少下游因子的分泌。结论:中药金银花、连翘、蒲公英可以抑制SV40 MES 13 NF-κB炎症信号通路的激活从而抑制SV40 MES 13的增殖并减少其下游因子的表达,该作用可能是清热解毒中药改善肾脏病炎性损伤的重要机制。
Objective：to the Heat- clearing and Detoxifying Herbs such as Honeysuckle, Weeping Forsythia as a research object, focusing on observation of the influence of the Heat - clearing and Detoxifying Herbs to NF - κB signaling pathway in SV40 MES 13 by TNF-α. discuss the effective mechanism of the Heat- clearing and Detoxifying Herbs to nephropathy, which providin. Methods： Detecting the effect of ten Heat - clearing and Detoxifying Herbs on the cell growth by MTT and select three herbs that with moderate toxic and wide role concentration range. The SV40 MES 13 induced by TNF - α were treated with the three herbs respectively and then MTT colorimetric assay was used to detect the proliferation of SV40 MES 13 ,the amounts of the cytoplasm p - IKBα,NF - κB p65 and of nuclear NF - κB p65 in SV40 MES 13, MCP - 1 and IL - 6 in cell supernatant were determined by ELISA. 2）. Results： ELISA results showed that the treatment groups decreased amounts of cytoplasm p -IKBot and nuclearNF - κBp65 （P 〈 0.01 ）, as well as the amounts of MCP - 1 and IL -6 in cell supernatant（P 〈 0.01 ）, honeysuckle had the strongest action in the three herbs with the same concentration. It mained that the Heat - clearing and Detoxifying Herbs could inhibite NF - KB activation in SV40 MES 13 and decrease the secretion of downstream factor. Conclusion：The Heat - clearing and Detoxifying Herbs can inhibite the proliferation of SV40 MES 13 and decrease the secretion of it＇ s downstream factor by inhibit Rel/NF - κB/IKB signal transduction pathway. It might be the mechanism of the Heat - clearing and Detoxifying Herbs against the inflammatory injury of kidney.
出处
《中国中西医结合肾病杂志》
2013年第12期1047-1050,共4页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
江苏省中医药局基金资助项目(No.LZ11067)
关键词
清热解毒
肾病
NF-ΚB
Heat - clearing and detoxifying herbs Nephropathy NF - κB
