作　　者： ; ; ; ;

机构地区： 中国中医科学院中医临床基础医学研究所

出　　处： 《中国实验方剂学杂志》 2020年第5期48-53,共6页

摘　　要： 目的:从小鼠小胶质瘤细胞(BV2)极化角度探索片仔癀(PTH)的抗炎作用机制。方法:将对数生长期的BV2细胞分为空白组,M1模型组[脂多糖(LPS)100μg·L^-1+干扰素-γ(IFN-γ)10μg·L^-1],M1片仔癀组[LPS 100μg·L^-1+IFN-γ10μg·L^-1+PTH 0.4 g·kg^-1],M2模型组[白细胞介素-4(IL-4)20μg·L^-1],M2片仔癀组[IL-420μg·L^-1+PTH0.4 g·kg^-1],给以相应处理。通过酶联免疫吸附测定(ELISA)检测细胞上清中肿瘤坏死因子-α(TNF-α),一氧化氮(NO),白细胞介素^-10(IL^-10),转化生长因子-β1(TGF-β1)的含量,使用实时荧光定量聚合酶链式反应(Real-time PCR)检测细胞中一氧化氮诱导合酶(i NOS),精氨酸^-1(Arg-1)mRNA的水平,蛋白免疫印迹法(Western blot)检测细胞中i NOS,p-STAT1,p-STAT3,Arg-1,p-STAT6蛋白表达情况。结果:与空白组比较,M1模型组上清TNF-α,NO含量显著升高(P<0.01),细胞i NOS mRNA,i NOS,p-STAT1,p-STAT3蛋白水平显著升高(P<0.01),而与M1模型组比较,M1片仔癀组上清TNF-α及NO的含量显著下调(P<0.01),细胞i NOS mRNA显著下调(P<0.01),i NOS,p-STAT1,p-STAT3蛋白水平也明显下调(P<0.05,P<0.01)。与空白组比较,M2模型组细胞培养上清中IL^-10及TGF-β1含量显著升高(P<0.01),细胞Arg-1 mRNA,Arg-1,p-STAT6蛋白水平显著升高(P<0.01)。与M2模型组比较,M2片仔癀组上清中IL-10及TGF-β1含量明显上调(P<0.05,P<0.01),细胞Arg-1mRNA水平,Arg-1及p-STAT6蛋白水平明显上调(P<0.05,P<0.01)。结论:片仔癀可以通过调节BV2的极化发挥抗炎作用。 Objective:To investigate the anti-inflammation mechanism of Pien Tze Huang(PTH)via regulating microglia polarization.Method:The experiment was divided into five groups,Blank,M1[lipopolysaccharide(LPS)100μg·L^-1+interferon-γ(IFN-γ)10μg·L^-1],M1-PTH group[LPS 100μg·L^-1+IFN-γ10μg·L^-1+PTH 0.4 g·kg^-1],M2 group[interleukin-4(IL-4)20μg·L^-1],and M2-PTH group[IL-420μg·L^-1+PTH 0.4 g·kg^-1].The concentration of nitric oxide(NO),tumor necrosis factor-α(TNF-α),interleukin^-10(IL^-10),and transforming growth factor-β1(TGF-β1)in the culture supernatant were measured by enzyme-linked immunosorbent assay(ELISA),the levels of inducible nitric oxide synthase(i NOS)and arginine^-1(Arg-1)mRNA were detected by real-time fluorescent quantitative polymerase chain reaction technique(Realtime PCR),and the expression levels of p-STAT1,p-STAT3,i NOS,p-STAT6,and Arg-1 were detected by Western blot.Result:The concentration of NO and TNF-αof the culture supernatant,the level of i NOS mRNA,as well as the level of p-STAT1,p-STAT3 and i NOS in M1 group,which were significantly increased(P<0.01).Compared with blank group,but the concentration of NO and TNF-αwere down-regulated(P<0.01),and i NOS mRNA(P<0.05),as well as the expression of i NOS,p-STAT1,and p-STAT3 was decreased(P<0.05,P<0.01)after the invention of PTH in M1-PTH group compared with M1 group.The concentration of IL^-10 and TGF-β1in the culture supernatant,the mRNA level of Arg-1,as well as the levels of p-STAT6 and Arg-1 were significantly increased in M2 group when compared with Blank group,addition to the concentration of IL^-10 and TGF-β1were up-regulated(P<0.05,P<0.01),and the expression of Arg-1 mRNA,the level of Arg-1,pSTAT6 were enhanced(P<0.05,P<0.01)in M2-PTH group compared with M2 group.Conclusion:PTH plays an anti-inflammatory role via regulating microglia polarization.

关 键 词： 片仔癀 小胶质细胞 极化 抗炎 中医药

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