参连复脉颗粒对脂多糖诱导的RAW264.7巨噬细胞极化的影响

Effect of Shenlian Fumai Granules on Lipopolysaccharide-induced Polarization of RAW264.7 Macrophages

目的观察参连复脉颗粒对脂多糖(LPS)诱导的RAW264.7巨噬细胞极化的影响,探讨其干预心房颤动的可能相关机制。方法使用LPS刺激RAW264.7巨噬细胞24 h构建巨噬细胞炎症模型。细胞分为正常对照组(空白血清)、模型组(LPS+空白血清)、中药组(LPS+参连复脉颗粒含药血清)、阿托伐他汀钙组(LPS+空白血清+阿托伐他汀钙)。采用四甲基偶氮唑盐(MTT)法检测巨噬细胞活性及增殖能力;采用荧光染色双标法观察巨噬细胞的极化状态;采用蛋白质印迹法(Western Blot)检测巨噬细胞白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)蛋白表达水平。结果与正常对照组比较,模型组细胞活力OD值升高(P<0.05),CD_(163)^(+)/CD_(68)^(+)比值降低(P<0.05),IL-6、TNF-α蛋白表达水平升高(P<0.05);与模型组比较,中药组和阿托伐他汀钙组细胞活力OD值降低(P<0.05),CD_(163)^(+)/CD_(68)^(+)比值升高(P<0.05),中药组IL-6、TNF-α蛋白表达水平降低(P<0.05);中药组细胞活力OD值、CD_(163)^(+)/CD_(68)^(+)比值、IL-6及TNF-α蛋白表达水平与阿托伐他汀钙组比较,差异均无统计学意义(P>0.05)。结论参连复脉颗粒含药血清可以抑制LPS诱导的巨噬细胞IL-6、TNF-α蛋白的分泌表达,抑制炎症反应,促进M1型巨噬细胞向M2型巨噬细胞转化。

Objective To observe the effect of Shenlian Fumai Granules on the polarization of RAW264.7 macrophages induced by lipopolysaccharide(LPS),and to explore the possible mechanism of its intervention in atrial fibrillation.Methods RAW264.7 macrophages were stimulated with LPS for 24 h to establish macrophage inflammation model.The cells were divided into normal control group(blank serum),model group(LPS+blank serum),traditional Chinese medicine group(LPS+Shenlian Fumai Granules containing serum),and atorvastatin calcium group(LPS+blank serum+atorvastatin calcium).The activity and proliferation of macrophages were detected by MTT assay.The polarization of macrophages was observed by immunofluorescence histochemical double-staining.The protein expressions of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in macrophages were detected by Western Blot.Results Compared with normal control group,the OD value of cell viability in model group was increased(P<0.05),the CD_(163)^(+)/CD_(68)^(+)ratio was decreased(P<0.05),and the protein expression levels of IL-6 and TNF-αwere increased(P<0.05).Compared with model group,the OD value of cell viability in traditional Chinese medicine(TCM)group and atorvastatin calcium group was decreased(P<0.05),the CD_(163)^(+)/CD_(68)^(+)ratio was increased(P<0.05),and the protein expression levels of IL-6,and TNF-αin the traditional Chinese medicine group were decreased(P<0.05).There were no significant differences in cell viability OD value,CD_(163)^(+)/CD_(68)^(+)ratio,IL-6,and TNF-αprotein expression levels between the traditional Chinese medicine group and the atorvastatin calcium group(P>0.05).Conclusion Shenlian Fumai Granules can inhibit the secretion and expression of IL-6,and TNF-αproteins in macrophages induced by LPS,inhibit the inflammatory response,and promote the transformation of M1 macrophages into M2 macrophages.