冠心平含药血清对LPS诱导的RAW264.7巨噬细胞极化的影响

Effects of Guanxinping-Containing Serum on RAW 264.7 Macrophages Polarization Induced by LPS

目的探讨冠心平(GXP)含药血清对LPS诱导的RAW264.7巨噬细胞极化的调控作用。方法RAW264.7巨噬细胞分为对照组、模型组、GXP低、中、高剂量组,100 ng/mL LPS刺激12 h诱导M1型极化模型,不同浓度GXP含药血清进行干预。ELISA法检测巨噬细胞上清肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)含量;流式细胞术检测M1、M2型巨噬细胞特征性表面标记分子CD86、CD206表达水平;qPCR法检测巨噬细胞CD86、CD206 mRNA表达情况。结果冠心平高剂量显著减少巨噬细胞TNF-α分泌(P<0.01),各剂量减少TGF-β1分泌(P<0.01);冠心平低、高剂量显著降低M1型巨噬细胞特征性分子CD86表达(P<0.05),显著升高M2型巨噬细胞特征性分子CD206表达(P<0.01);同时冠心平CD206 mRNA表达显著增加(P<0.05)。结论冠心平含药血清能够抑制LPS诱导的巨噬细胞M1型极化而促进M2型极化,这或许是冠心平治疗动脉粥样硬化的潜在机制之一。

OBJECTIVE To investigate the effects of Guanxinping(GXP)on RAW264.7 macrophage polarization induced by LPS.METHODS Macrophages were divided into control group,model group and low-,mid-,high-dose of GXP groups.Cells were starved for 24 hours before intervention with drug-containing serum of rats.The characteristic molecules of macrophage polarization in cell supernatant,including tumor necrosis factorα(TNF-α)and transforming growth factorβ(TGF-β)were detected by ELISA;the expression levels of M1 and M2 macrophage surface molecules CD86 and CD206 were detected by flow cytometry;the expression of mRNA of CD86 and CD206 were detected by qPCR.RESULTS Compared to the model group,high-dose of GXP decreased the secretion of TNF-α(P<0.01);low-,mid-,high-dose of GXP decreased the secretion of TGF-β1(P<0.01)in macrophage supernatant.Low-and high-dose of GXP decreased the expressions of CD86(P<0.05)and increased the expression of CD206(P<0.01).There were no significant difference in the mRNA expression of CD86 between the GXP groups and model group,but GXP increased the mRNA expression of CD206.CONCLUSION GXP can inhibit the polarization of macrophages towards M2,which may be one of the mechanisms of GXP in the treatment of atherosclerosis.