慢性应激小鼠脑内STING-TBK1-IRF3通路相关蛋白的表达及意义

Expression and role of STING-TBK1-IRF3 signaling pathway in the brain of mice with chronic stress

目的探索慢性应激小鼠脑内干扰素刺激因子(stimulator of interferon gene,STING)-TANK结合激酶1(TANK-binding kinase 1,TBK1)-干扰素调节因子3(interferon regulatory factor 3,IRF3)信号通路相关蛋白的表达变化及意义。方法将小鼠分为对照(control,CON)组与束缚应激(chronic restraint stress,RST)组,RST组小鼠给予慢性束缚应激刺激(每天6 h,共14 d)后,采用q RT-PCR、组织免疫荧光共标染色与Western blotting方法检测两组小鼠脑内促炎因子CCL2、CXCL10、IL-1β、IL-6、IL-10、TNFαm RNA与STING、TBK1、p-TBK1、IRF3、p-IRF3的蛋白表达变化。结果q RT-PCR实验结果显示,与CON组相比,RST组小鼠脑内促炎因子m RNA表达显著升高(P均<0.05);组织免疫荧光共标染色实验结果显示,STING与小胶质细胞标记物Iba-1高度共标,RST组小鼠脑内STING表达降低;Western blotting结果显示,STING、p-TBK1、p-IRF3蛋白表达均显著降低(P均<0.01)。结论慢性束缚应激小鼠脑内存在神经炎症反应,STING-TBK1-IRF3通路被显著抑制。

Objective To explore the expression and role of stimulator of interferon gene(STING)-TANKbinding kinase 1(TBK1)-interferon regulatory factor 3(IRF3)signaling pathway in the brain of chronic stress mice.Methods Mice were divided into control(CON)group and chronic restraint stress(RST)group.Mice in the RST group were given chronic restraint stress stimulation(6 hours per day,14 days).After 14 days,the m RNA expressions of pro-inflammatory cytokines CCL2,CXCL10,IL-1β,IL-6,IL-10,and TNFαin the brain were detected and analyzed by q RT-PCR;protein expression of STING,TBK1,p-TBK1,IRF3,and p-IRF3 were detected and analyzed by immunofluorescence staining and Western blotting.Results Compared to the CON group,the m RNA expressions of pro-inflammatory cytokines in the RST group were significantly increased(P<0.05).STING and microglia marker Iba-1 were highly co-located and the expression of STING was decreased as detected by immunofluorescence staining.Moreover,the protein expressions of STING,p-TBK1,and p-IRF3 were significantly decreased(all P<0.01).Conclusion Chronic restraint stress triggers a neuroinflammatory response and the STING-TBK1-IRF3 pathway in the brain of the RST mice is significantly inhibited.