硫化氢通过JAK2/STAT3通路抑制脂多糖诱导的小胶质细胞M1型极化

Hydrogen sulfide inhibits the M1 polarization of microglia induced by lipopolysaccharide through the JAK/STAT3 pathway

目的观察硫化氢(H2S)对脂多糖(LPS)诱导的小胶质细胞极化的影响并探讨JAK2/STAT3信号通路是否介导其作用。方法将培养的N9小胶质细胞随机分为对照组、LPS组、硫氢化钠(NaHS)组、LPS+NaHS组、蛋白酪氨酸激酶(JAK2)/信号转导和转录激活因子(STAT3)抑制剂α-氰基-(3,4-羟基)N-苄苯乙烯胺(AG-490)组、LPS+NaHS+AG-490组共6组,每组设3个复孔。采用MTT法检测各组细胞活力,采用ELISA法检测各组胶质细胞及培养上清液中白细胞介素1β(IL-1β)、IL-6、IL-4 IL-10水平,采用Western-blot检测各组细胞精氨酸酶1(Arg1)、诱导型一氧化氮合酶(iNOS)、磷酸化JAK2(p-JAK2)和磷酸化STAT3(p-STAT3)蛋白表达。结果与对照组比较,LPS组细胞Arg1蛋白及IL-4和IL-10表达水平增加,而iNOS蛋白、IL-1β、IL-6、p-JAK2蛋白和p-STAT3蛋白表达降低,p-JAK2/t-JAK和p-STAT3/t-STAT3比值降低(均P<0.05)。与LPS组比较,LPS+NaHS组细胞Arg1蛋白及IL-4和IL-10表达水平增加,而iNOS蛋白、IL-1β、IL-6、p-JAK2蛋白和p-STAT3蛋白表达降低,p-JAK2/t-JAK2和p-STAT3/t-STAT3比值降低(均P<0.05)。JAK2/STAT3信号通路抑制剂AG-490可减弱NaHS的作用。结论H2S可抑制LPS诱导的小胶质细胞M1型极化,其机制可能是通过JAK2/STAT3信号通路实现。

Objective To investigate the effect of hydrogen sulfide(H2S)on the microglial polarization induced by lipopolysaccharide(LPS)and whether the JAK2/STAT3 signaling pathway mediates its action.Methods Cultured N9 microglia were randomly divided into six groups:a control group,an LPS group,an NaHS group,an LPS+NaHS group,a protein tyrosine kinase(JAK2)/signal transducer group,an activator of transcription(STAT3)inhibitorα-cyano-(3,4-hydroxy)n-benzyl-styramine(AG-490)group,and an LPS+NaHS+AG-490 group.Each group had three multiple pores.MTT assay was used to detect cell viability.The levels of IL-1β,IL-6,IL-4,and IL-10 in glial cells and culture supernatant were detected by ELISA.Western Blot was used to detect the expression of inducible nitric oxide synthetase(iNOS),arginase 1(Arg1),phosphorylated janus kinase 2(p-JAK2),phosphorylated signal transducer,and activator of transcription 3(p-STAT3).Results Compared with the control group,Arg1 protein expression,IL-4,and IL-10 levels were significantly decreased,while iNOS protein expression,IL-1βand IL-6 levels,p-JAK2 and p-STAT3 protein expression,p-JAK2/t-JAK2 and p-STAT3/t-STAT3 ratios were significantly increased in the LPS group(all P<0.05).Compared with the LPS group,Arg1 protein expression,IL-4 and IL-10 levels in the LPS+NaHS group were significantly increased,while iNOS protein expression,IL-1βand IL-6 levels,p-JAK2 and p-STAT3 protein expression,p-JAK2/t-JAK2 and p-STAT3/t-STAT3 ratios were significantly decreased in the LPS+NaHS group(all P<0.05).AG-490,a JAK2/STAT3 signal pathway inhibitor,attenuated the effect of NaHS.Conclusions Hydrogen sulfide inhibits the M1 polarization of microglia induced by LPS,which may be realized through the JAK2/STAT3 signaling pathway.