侯氏黑散化学成分对脂多糖诱导BV2细胞活化中炎性反应因子分泌平衡的影响  

Effects of chemical components of HSHS on the balance of inflammatory cytokines in LPS-stimulated BV2 cells

作　　者：常佳慧[1] 赵晖[1] 王蕾[1] 张弛 陆跃[1] 张秋霞[1] Chang Jiahui Zhao Hui Wang Lei Zhang Chi Lu Yue Zhang Qiuxia(Collateral Disease Research Unit, TCM School, Capital Medical University, Beijing 100069, China Journal Center, Beijing University of Chinese Medicine, Beijing 100029, China)

机构地区：[1]首都医科大学中医药学院中医络病研究北京市重点实验室,北京100069 [2]北京中医药大学期刊中心,北京100029

出　　处：《首都医科大学学报》2017年第2期213-219,共7页Journal of Capital Medical University

基　　金：国家自然科学基金(81373526);北京市自然科学基金(7102014;7122018);北京市属高等学校高层次人才引进与培养计划-长城学者(CIT&TCD20140329)~~

摘　　要：目的探究侯氏黑散方中绿原酸(chlorogenic acid,CGA)、木犀草苷(cynaroside,CYN)、木犀草素(luteolin,LUT)、人参皂苷Rg1(ginsenoside Rg1,GS-Rg1)对脂多糖(lipopolysaccharide,LPS)诱导小胶质细胞炎性反应因子表达的影响。方法实验分对照组、模型组、CGA、CYN、LUT、GS-Rg1处理组。利用LPS诱导BV2细胞过度活化,建立脑缺血后小胶质细胞炎性反应损伤体外模型;CCK-8法检测各组细胞活性;Griess法检测各组细胞上清中一氧化氮(nitric oxide,NO)的量;酶联免疫法(enzyme linked immunosorbent assay,ELISA)检测各组细胞上清中肿瘤坏死因子(tumor necrosis factor-α,TNF-α)、白介素-6(interleukin-6,IL-6)、白介素-10(interleukin-10,IL^(-1)0)、转化生长因子-β_1(transforming growth factor-β_1,TGF-β_1)的量;蛋白印迹法(Western blotting,WB)检测各组细胞p-p65、p-IκBα蛋白表达。结果与模型组相比,不影响细胞生存活性的情况下,CGA、CYN、LUT、GS-Rg1可显著降低细胞上清中促炎因子NO、TNF-α、IL-6浓度;GS-Rg1可显著升高抑炎因子IL^(-1)0、TGF-β_1浓度;CGA、CYN、LUT、GS-Rg1可显著降低细胞中p-p65、p-IκBα蛋白浓度。结论 CGA、CYN、LUT、GS-Rg1分别从促炎、抑炎两方面调节脑缺血后炎性反应相关因子分泌的平衡,其机制与核转录因子-κB(nuclear factor-κB,NF-κB)通路活化有关。Objective To investigate the influence of chlorogenic acid,cynaroside,luteolin,ginsenoside Rg1 on expression of inflammatory cytokines.Methods The experiment was divided into six groups： control,model,chlorogenic acid,cynaroside,luteolin and ginsenoside Rg1 groups.The mimic ischemia injured microglia model was induced by LPS.The cyto-activity was detected via cell count kit.The NO content was determined by Griess Reagent.Contents of TNF-α、IL-6、IL-10 and TGF-β1 were detected by ELISA.The expression levels of p-p65 and p-IκBα were detected by Western blotting.Results Compared with those results of model group,chlorogenic acid,cynaroside,luteolin and ginsenoside Rg1 groups could significantly inhibit the release of NO,and decrease the content of TNF-α and IL-6,ginsenoside Rg1 markedly elevated the TGF-β1 level without influencing the cell survival.Chlorogenic acid,cynaroside,luteolin and ginsenoside Rg1 groups could decrease the expression of p-p65,p-IκBα.Conclusion The findings demonstrated that chlorogenic acid,cynaroside,luteolin and ginsenoside Rg1 played regulating roles in balancing ischemia injured microglia homeostasis via promoting anti-inflammatory cytokines as well as inhibiting the inflammatory cytokines.The therapeutic roles in the inflammatory reaction of cerebral ischemia which perhaps worked through the activation of NF-κB signaling pathway.

关 键 词：侯氏黑散 绿原酸 木犀草苷 木犀草素 人参皂苷RG1 小胶质细胞 炎性反应因子 NF-κB信号通路 

分 类 号：R289.5[医药卫生—方剂学]