SOCS3在内毒素诱导神经胶质细胞耐受模型中的变化

摘要

Objective To study the changes of SOCS3 expression in LPS-induced mice glia cell tolerant(majorly referring to astrocytes and microglia).Methods Mice astrocytes and microglia were purified in vitro. Then the astrocytes and microglia were pretreated with 0.01ug /ml of LPS for 18 hours,and received the second times of treatment with 1ug/ml of LPS for 24 hours to induce LPS tolerance. Non-treated group,0.01ug/ml of LPS treated group,1ug/ml of LPS treated-group were also designed as control. After the treatment,the cells and cultured supernatants were harvested for determination of IL-6 by ELISA,SOCS3 expression by western blot.Results IL-6 levels were increased with LPS treatment(P<0.05). In astrocytes,IL-6 level was higher in LPS-pretreated group than that in lug/ml of LPS-treated group(P<0.05). LPS pretreatment tended to enhance the expression of SOCS3 protein.Conclusion SOCS3 pathway can involve in the re-adaption LPS tolerance in the neuroglia,which is one of the negative modulator of cellular signal in LPS tolerance of central nervous system.%目的 探讨内毒素(脂多糖,lipopolyssacride,LPS)体外诱导小鼠大脑皮质神经胶质细胞耐受模型中细胞因子信号转导抑制因子(Suppressor of Cytokine Signaling 3,SOCS3)信号的变化.方法 分离纯化新生小鼠大脑皮质星型胶质细胞和小胶质细胞,采用LPS进行预刺激(先采用0.01μg/ml LPS刺激18h后,再用1μg/ml LPS刺激24h)诱导LPS耐受,同时设置未刺激组、0.01μg/ml LPS和1μg/ml LPS单次刺激组对照.收获细胞与上清液,采用ELISA法检测星型胶质细胞与小胶质细胞IL-6水平,Western blot检测SOCS3蛋白表达水平.结果 LPS预刺激组产生IL-6水平与未刺激对照组,0.01μg/ml单次刺激组、1μg/ml单次刺激组相比水平升高,其中星型胶质细胞与1μg/ml单次刺激组比较升高明显(P<0.05);星型胶质细胞与小胶质细胞预激组SOCS3表达均有升高趋势.结论 SOCS3-IL-6信号轴与中枢神经细胞群LPS耐受效应密切相关,参与中枢LPS耐受细胞负控信号的重新调整.