摘要
目的探讨谷氨酸对HT22细胞线粒体自噬和细胞凋亡的影响,并评估虾青素预处理的保护作用及其分子机制。方法用谷氨酸及虾青素处理HT22细胞,通过蛋白印迹及聚合酶联反应等评估其对线粒体自噬的影响。结果谷氨酸处理显著抑制线粒体初级自噬(PINK1、Parkin、pULK1ser555和LC3Ⅱ)和次级自噬(LAMP1和Rab7),上调cleaved Caspase-3的表达(P<0.05)。虾青素预处理减少细胞凋亡,恢复了线粒体自噬,PINK1、Parkin、pULK1ser555和LC3Ⅱ的表达水平上升(分别为2.3倍、2.6倍、83.3%及81.1%)(P<0.05),该作用被自噬抑制剂BafA1阻断。此外,谷氨酸抑制Nrf2核内转移和NLRX1表达,而预处理显著促进Nrf2的核内转移并上调NLRX1,分别上调25.8%、33.2%。生物信息学分析显示NLRX1启动子区域含有3个Nrf2结合位点,提示Nrf2通过调控NLRX1转录活性发挥作用。结论文章首次揭示虾青素通过Nrf2/NLRX1通路激活线粒体自噬,展现神经保护作用。
Objective To explore the effects of glutamate on mitophagy and apoptosis in HT22 cells and evaluate the protective effects and molecular mechanisms of astaxanthin pretreatment.Methods HT22 cells were treated with glutamate and astaxanthin.The effects on mitophagy were assessed using Western Blot and PCR.Results Glutamate treatment significantly inhibited primary mitophagy(PINK1,Parkin,pULK1ser555 and LC3II)and secondary mitophagy(LAMP1 and Rab7)while upregulating cleaved Caspase-3 expression.Astaxanthin pretreatment notably reduced apoptosis and restored mitophagy,the expression levels of PINK1,Parkin,pULK1Ser555 and LC3II were significantly upregulated(by 2.3-fold,2.6-fold,83.3%and 81.1%respectively,P<0.05),but this effect was blocked by the autophagy inhibitor BafA1.Additionally,glutamate suppressed Nrf2 nuclear translocation and NLRX1 expression,whereas astaxanthin promoted Nrf2 nuclear translocation and increased NLRX1 expression by 25.8%and 33.2%,respectively.Bioinformatics analysis revealed three Nrf2 binding sites in the NLRX1 promoter region,suggesting that Nrf2 may regulate NLRX1 transcriptional activity.Conclusions The study demonstrates that astaxanthin exhibited potential neuroprotective effect by activating mitophagy through the Nrf2/NLRX1 pathway.
作者
温小军
肖丽姣
WEN Xiaojun;XIAO Lijiao(Department of Neurology,Guangzhou First People’s Hospital,the Second Affiliated Hospital,School of Medicine,South China University of Technology,Guangzhou 510180,China)
出处
《广州医药》
2025年第7期975-981,共7页
Guangzhou Medical Journal
基金
广东省医学科研基金(A2020281)
广州市卫健委西医类一般引导项目(20231A011014)。
