摘要
目的研究中药地黄饮子抑制脂多糖(LPS)诱导的小胶质细胞炎症反应的作用机制。方法子组(14.5 g·kg-1)、米诺环素组(150 mg·kg-1)。地黄饮子组和米诺环素组予以10%的含药血清干预,空白组和模型组给予同等浓度的空白血清。明场下观察细胞形态的变化,实时荧光定量PCR检测促炎性因子IL-1β、IL-6、TNF-αmRNA表达水平,Western blot检测PPARγ蛋白表达和NF-κB p65活化水平,细胞免疫荧光检测NF-κB p65入核情况。结果与空白组比较,模型组促炎性因子IL-1β、IL-6、TNF-α的mRNA水平均显著升高(P<0.01);PPARγ蛋白水平明显降低,NF-κB磷酸化水平明显增高,差异均有统计学意义(P<0.01),LPS刺激使BV2细胞核内NF-κB含量明细增多;给予地黄饮子或米诺环素含药血清干预后,与模型组比较,炎性因子IL-1β、IL-6、TNF-α的mRNA水平均显著降低(P<0.05),PPARγ蛋白水平明显升高(P<0.05),NF-κB磷酸化水平和入核含量明显减少(P<0.05)。结论地黄饮子可能通过调控PPARγ/NF-κB信号通路抑制LPS诱导的BV2细胞炎症反应。
Objective To investigate the inhibitory effect of Dihuang-Yinzi Decoction on LPS induced inflammatory reaction and its specific mechanism.Methods The inflammation model of BV2 cell was induced by LPS.The cultured cells were divided into:blank group,model group,Dihuang-Yinzi group,and minocycline group.The Dihuangyinzi group and minocycline group were intervened with 10%medicated serum,and the blank group and model group were given the same concentration of blank serum.Real time PCR was used to detect the mRNA expression of proinflammatory factors IL-1β,IL-6 and TNFα.Western blotting was used to detect the expression of PPARγand the activation of NF-κBp65 in BV2 cells.Immunofluorescence was used to detect the nuclear entry of NF-κBp65.Results Compared with the blank group,the mRNA levels of proinflammatory cytokines IL-1β,IL-6 and TNFαin the LPS model group were significantly increased(P<0.01).PPARγprotein level was significantly decreased,and NF-κB phosphorylation level was significantly increased(P<0.01).LPS stimulation increased the nuclear content of NF-κB in BV2 cells.After intervention with Dihuangyinzi or minocycline medicated serum,the mRNA levels of inflammatory factors IL-1β,IL-6 and TNF-αwere significantly lower than those in the model group(P<0.05),the protein level of PPARγwas significantly higher than that in the model group(P<0.05),and the phosphorylation level and nuclear content of NF-κB were significantly lower than those in the model group(P<0.05).Conclusion Dihuangyinzi can inhibit LPS-induced inflammatory response of BV2 cells,and its specific mechanism may be related to thePPARγ/NF-κB signaling pathway.
作者
江建锋
白强
贺春香
李泽
宋祯彦
成绍武
Jiang Jianfeng;Bai Qiang;He Chunxiang;Li Ze;Song Zhenyan;Cheng Shaowu(The Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410005,China;Key Lab of Hunan Province for Prevention and Treatment of Cardio-cerebral Disease with Integrated Traditional Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2021年第5期1610-1616,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会面上项目(81774129):基于“痰浊瘀血”理论从蛋白质异戊二烯化修饰探讨当归芍药散改善AD认知功能的机制,负责人:成绍武
湖南省科学技术厅湖南创新型省份建设专项(2019RS1064):2019年湖湘高层次人才聚集工程-创新人才,负责人:成绍武
湖南省自然科学基金委员会科卫联合项目(2020JJ8023):地黄饮子通过NFκB-NLRP3复合体/STAT6-PPARγ调控小胶质细胞表型极化保护神经元的作用,负责人:江建锋。
