摘要
目的:探讨壮骨健膝方对巨噬细胞经典活化型(M1)/替代活化型(M2)极化的影响及作用机制.方法:佛波酯诱导人单核白血病细胞分化为非活化巨噬细胞(M0),脂多糖刺激M0复制M1极化模型,将细胞分为空白组(M0+空白血清)、模型组(M1+空白血清)、抑制剂组(M1+NF-κB抑制剂+空白血清)、壮骨健膝方组(M1+含药血清).干预24 h后,检测各组M1标志分子诱导型一氧化氮合酶(iNOS)、主要组织相容性复合体Ⅱ(MHC-Ⅱ)、单核细胞趋化蛋白1(MCP-1),M2标志分子分化簇206、163、209(CD206、163、209)mRNA表达,炎症介质白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α),抗炎因子IL-4、IL-10水平及mRNA表达,核因子κB(NF-κB)信号通路中节点蛋白核内NF-κBp65、NF-κBp50及NF-κB抑制蛋白α(IKBα)表达.结果:模型组M1标志分子、炎症介质及NF-KBp65、NF-κBp50蛋白均高于空白组(P＜0.05),CD206、IL-4 mRNA及IκBα蛋白,IL-4水平均低于空白组(P＜0.05);抑制剂及壮骨健膝方组M1标志分子、炎症介质及NF-κBp65、NF-κBp50蛋白均低于模型组(P＜0.05),M2标志分子、抗炎因子及IκBα蛋白均高于模型组(P＜0.05);壮骨健膝方组MHC-Ⅱ、MCP-1 mRNA及炎症介质均低于抑制剂组(P＜0.05),CD206、CD209、IL-10 mRNA及IL4、IL-10水平均高于抑制剂组(P＜0.05).结论:壮骨健膝方可抑制脂多糖诱导的巨噬细胞M1极化,促进M2极化,其机制可能与降低细胞中NF-κB信号通路活性有关.
Abstract
Objective:To investigate the effect and mechanism of Zhuanggu Jianxi Formula(ZGJXF)on the polarization of macro-phages into classical activation(M1)and alternative activation(M2)types.Methods:Phorbol 12-myristate 13-acetate was used to induce differentiation of human monocytic leukemia cells into non-activated macrophages(M0).Lipopolysaccharide was used to stimulate M0 to replicate the M1 polarization model.Cells were divided into a blank group(M0+blank serum),a model group(M1+blank serum),an inhibitor group(M1+NF-κB inhibitor+blank serum),and a ZGJXF group(M1+medicated serum).After 24 hours of intervention,various indicators were measured,including M1 markers inducible nitric oxide synthase(iNOS),major histo-compatibility complex Ⅱ(MHC-Ⅱ),monocyte chemotactic protein 1(MCP-1),M2 markers CD206,CD163,CD209 mRNA expres-sion,pro-inflammatory factors interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),anti-inflammatory factors IL-4,IL-10 levels and mRNA expression,and key proteins in the NF-κB signaling pathway,such as nuclear NF-KBp65,NF-KBp50,and NF-κB inhib-itor α(IκBα).Results:In the model group,M1 markers,pro-inflammatory factors,and NF-KBp65,NF-KBp50 proteins were signifi-cantly higher than those in the blank group(P＜0.05),while CD206,IL-4 mRNA,IKBα protein,and IL-4 levels were lower than those in the blank group(P＜0.05).In the inhibitor and ZGJXF groups,M1 markers,pro-inflammatory factors,and NF-KBp65,NF-KBp50 proteins were lower than those in the model group(P＜0.05),while M2 markers,anti-inflammatory factors,and IκBα pro-tein were higher than those in the model group(P＜0.05).The ZGJXF group had lower levels of MHC-Ⅱ,MCP-1 mRNA,and pro-inflammatory factors than the inhibitor group(P＜0.05),and higher levels of CD206,CD209,IL-10 mRNA,IL-4,and IL-10(P＜0.05).Conclusion:ZGJXF can inhibit lipopolysaccharide-induced M1 polarization of macrophages and promote M2 polarization.The mechanism may be related to the reduction of the activity of the NF-κB signaling pathway in cells.
维普
万方数据