四逆散对脂多糖诱导的RAW264.7细胞极化的影响

Effect of Sinisan on Macrophage Polarization of RAW264.7 Cells Induced by Lipopolysaccharide

目的:探讨四逆散(Sinisan,SNS)对脂多糖(LPS)诱导的RAW264. 7巨噬细胞极化的调控作用。方法:RAW264. 7分为5组,分别为空白组,模型组,SNS低、中、高质量浓度组(10,20,40 mg·L^(-1));以LPS(100μg·L^(-1))刺激的RAW264. 7细胞为体外模型,使用不同质量浓度的SNS提前干预细胞,噻唑蓝(MTT)比色法检测不同质量浓度SNS对RAW264. 7细胞增殖的影响;光学显微镜下观察各组细胞分化程度;酶联免疫吸附测定(ELISA)检测细胞培养基上清中M1极化因子肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6),白细胞介素-1β(IL-1β)以及M2极化因子白细胞介素-10(IL-10)的含量;实时荧光定量聚合酶链式反应(Real-time PCR)检测RAW264. 7细胞M1极化因子TNF-α,IL-6以及M2极化因子IL-10,精氨酸酶-1(Arg^(-1))的mRNA水平。结果:与空白组比较,模型组促进细胞增殖(P <0. 05),刺激M1极化因子TNF-α,IL-6,IL-1β的释放和上调TNF-α,IL-6的mRNA含量(P <0. 01),减少M2极化因子IL-10释放和IL-10,Arg^(-1)的mRNA水平(P <0. 05,P <0. 01)。与空白组比较,SNS对RAW264. 7细胞的活性没有影响。与模型组比较,SNS可抑制LPS诱导的细胞增殖(P <0. 05),减少LPS刺激的细胞分化,减少M1极化因子TNF-α,IL-6,IL-1β的释放和TNF-α,IL-6的mRNA含量(P <0. 05,P <0. 01),并增加M2极化因子IL-10释放和IL-10,Arg^(-1)的mRNA水平(P <0. 05,P <0. 01)。结论:SNS可抑制LPS诱导的RAW264. 7细胞炎症,其机制可能与调控巨噬细胞M1/M2表型极化平衡相关。

Objective: To investigate the regulatory effect of Sinisan( SNS) on the polarization of RAW264. 7 macrophages induced by lipopolysaccharide( LPS). Method: RAW264. 7 cells stimulated by LPS were used as the in vitro model. The cells were intervened with the different concentrations of SNS in advance. The effects of different concentrations of SNS on the proliferation of RAW264. 7 cells were detected by methyl thiazolyl tetrazolium( MTT) colorimetry. The degree of cell differentiation was detected by enzyme-linked immuno sorbent assay( ELISA) method. The contents of M1 polarization factors tumor necrosis factor-α( TNF-α),interleukin-6( IL-6),interleukin^-1β( IL-1β) and M2 polarization factors interleukin^-10( IL-10) in cell culture supernatant were detected by ELISA method,mRNA levels of M1 polarization factors TNF-α,IL-6 and M2 polarization factors IL-10,arginase-1( Arg^-1) were detected by Real-time fluorescence quantitative polymerase chain reaction( Realtime PCR) method. Result: SNS had no effect on the cell viability of RAW264. 7 cells,inhibited LPS-induced cell proliferation,decreased LPS-stimulated cell differentiation,down-regulated M1 polarizing factors TNF-α,IL-6,IL-1β release and TNF-α,IL-6 mRNA levels,and increased the release of IL-10 and mRNA levels of IL-10 and Arg^-1. Conclusion: SNS inhibits the inflammation of RAW264. 7 cells induced by LPS,and its mechanism may be related to the regulation of polarization balance of M1/M2 macrophages.