芍药苷通过JAK2/STAT3信号通路抑制高糖诱导的RAW264.7巨噬细胞激活

目的探讨芍药苷(paeoniflorin,PF)抑制高糖(high glucose,HG)诱导的RAW264.7巨噬细胞激活是否通过JAK2/STAT3信号通路。方法体外HG激活巨噬细胞RAW264.7, PF及JAK2/STAT3干扰RNA(siRNA)进行干预,分别检测巨噬细胞的增殖、趋化、炎症因子表达分泌、JAK2和STAT3蛋白表达及其磷酸化水平。结果在HG刺激下,巨噬细胞趋化功能增强,诱生型一氧化氮合酶(iNOS)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)及单核细胞趋化因子1(MCP-1)的mRNA表达,以及细胞培养基中TNF-α、IL-1β、MCP-1分泌水平均增加(P<0.05,P<0.01),JAK2、STAT3蛋白磷酸化表达明显增加(P<0.05)。JAK2/STAT3基因沉默可抑制HG刺激下iNOS和炎症因子(TNF-α、IL-1β、MCP-1)mRNA水平和细胞培养液中炎症因子的分泌,抑制JAK2、STAT3蛋白磷酸化水平(P<0.05,P<0.01)。PF能明显下调HG诱导的巨噬细胞趋化迁徙、炎症因子表达分泌及JAK2、STAT3蛋白磷酸化水平(P<0.01)。结论 HG可通过诱导JAK2/STAT3信号通路刺激巨噬细胞激活,PF抑制巨噬细胞激活的机制主要与抑制JAK2/STAT3信号通路相关。

Paeoniflorin inhibits activation of Raw 264. 7 macrophages induced by high glucose via JAK2/STAT3 signaling pathway

Aim To investigate whether paeoniflorin(PF)inhibits the activation of RAW264.7macrophages induced by high glucose(HG)via JAK2/STAT3signaling pathway.Methods RAW264.7macrophages were cultured in vitro.HG was used as the stimulator.PF and JAK2/STAT3gene silencing interfered with macrophage activation.All cells were divided into nine groups:LG,HG,HG+PF,LG+JAK2siRNA,JAK2siRNA+HG,JAK2siRNA+HG+PF,LG+STAT3siRNA,STAT3siRNA+HG,STAT3siRNA+HG+PF.Macrophage proliferation,chemotaxis,expression and secretion of inflammatory cytokines,JAK2and STAT3protein expression and the phosphorylation levels were detected respectively.Results Compared with LG group,in HG group,the macrophages’chemotactic function significantly increased(P<0.05),the mRNA expression of iNOS,TNF-α,IL-1βand MCP-1and the secretion of inflammatory cytokines(TNF-α,IL-1β,MCP-1)in cell culture medium were markedly elevated(P<0.05,P<0.01),and the phosphorylation of JAK2and STAT3protein significantly increased(P<0.05).JAK2and STAT3gene silencing could suppress the HG-induced the mRNA expression of iNOS,TNF-α,IL-1βand MCP-1and the secretion inflammatory cytokines in cell culture medium as well as the phosphorylation of JAK2and STAT3protein(P<0.05,P<0.01).PF could significantly inhibit HG-induced macrophage chemokine and migration,inflammatory cytokine production and secretion,and JAK2,STAT3protein phosphorylation(P<0.01).Conclusion HG can stimulate macrophage activation by inducing JAK2/STAT3signaling pathway,and PF inhibits the activation of Raw264.7macrophages induced by high glucose via JAK2/STAT3pathway suppression.