目的 探究依达拉奉对LPS激活的小胶质细胞Sirt3和MAPKs通路相关蛋白的作用及其调控机制。 方法 采用Western blot和免疫荧光双标染色检测LPS激活的BV2小胶质细胞中Sirt3和MAPKs通路相关蛋白ERK1/2、P38、JNK及p-ERK1/2、p-P38和p-JNK的表达；检测ERK1/2通路抑制剂处理后p-ERK1/2和Sirt3的表达。 结果 LPS激活的BV细胞中Sirt3、p-ERK1/2、p-P38和p-JNK表达显著增高，依达拉奉能促进Sirt3和p-ERK1/2过表达，并降低p-P38和p-JNK表达；ERK1/2抑制后p-ERK1/2和Sirt3的蛋白表达降低。依达拉奉可促进激活的BV2细胞中Sirt3和p-ERK1/2过表达，并下调过表达的p-P38和p-JNK。 结论 依达拉奉可调节BV2细胞中MAPKs信号通路并促进Sirt3生成，可能经ERK/Sirt3通路发挥抗炎作用。
Abstract
Objective To investigate the effect of edaravone on Sirt3 and MAPKs pathway-related proteins in LPS-activated microglia and its regulatory mechanism. Methods Western Blot and immunofluorescence were used to detect the effect of edaravone on the protein expression of Sirt3 and MAPKs pathway-related proteins ERK1/2, P38, JNK as well as their phosphorylated forms p-ERK1/2, p-P38 and p-JNK in the activated BV2 cells. Western Blot techniques were applied to detect the effects of ERK1/2 pathway inhibitor on the expressions of p-ERK1/2 and Sirt3. Results The expressions of Sirt3, p-ERK1/2, p-P38 and p-JNK in microglia were significantly increased after BV2 cells activation. Edaravone promoted the expression of Sirt3 and p-ERK1/2 and downregulated the expression of p-JNK and p-P38. The protein expression of Sirt3 and p-ERK1/2 decreased after co-treatment with edaravone and ERK1/2 inhibitor. Edaravone promoted the overexpression of Sirt3 and p-ERK1/2 and downregulated the expression of p-JNK and p-P38. Conclusions Edaravone can regulate MAPKs signaling pathway and promote Sirt3 production in BV2 cells, possibly regulates neuroinflammatory responses via ERK/Sirt3 signaling pathway.
关键词
依达拉奉； /
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Sirt3； /
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小胶质细胞； /
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MAPKs信号通路
Key words
Edaravone; /
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Sirt3; /
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Microglia; /
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MAPKs signaling pathway
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基金
国家自然科学基金（81960223）；云南省科技厅昆明医科大学联合专项（202301AY070001-163）；云南省大学生创新性实验计划项目；昆明医科大学大学生创新性实验计划项目（2022JXD301）
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