BNIP3L介导的线粒体自噬在镉致大鼠大脑皮质神经元凋亡中的作用OA北大核心
The role of BNIP3L-mediated mitophagy in cadmium-induced apoptosis in rat cerebral cortical neurons
为探究线粒体自噬受体Bcl-2/腺病毒E1B 19 kDa相互作用蛋白3样(BNIP3L)介导的线粒体自噬对镉致大鼠大脑皮质神经元凋亡的调控作用,利用RNA干扰技术建立大鼠大脑皮质神经元BNIP3L基因沉默模型,并经10 μmol/L镉处理12 h,Western blot检测BNIP3L、帕金森氏蛋白2(Parkin)和线粒体凋亡通路相关蛋白cleaved caspase-9、cleaved caspase-3的表达水平,免疫荧光染色检测BNIP3L与线粒体标志蛋白TOMM20共定位,透射电镜检测细胞内线粒体自噬体数目,FITC Annexin V染色检测细胞凋亡水平.结果:大鼠大脑皮质神经元经镉处理12 h后,BNIP3L蛋白表达水平极显著升高(P＜0.01),BNIP3L与TOMM20共定位增加,沉默BNIP3L极显著抑制镉致Parkin线粒体转位(P＜0.01),抑制镉致细胞内线粒体自噬体形成,极显著促进镉致caspase-9、caspase-3激活和神经元凋亡(P＜0.01).结果表明,BNIP3L介导的线粒体自噬可抑制镉致大鼠大脑皮质神经元凋亡.
In order to determine the regulatory effect of BNIP3L-mediated mitophagy on apoptosis induced by cadmium(Cd),the rat cere-bral cortical neuron BNIP3L gene silencing model was established,in this study,by RNA interference technique,and then the model was treated with 10μmol/L Cd for 12 h.Next,the expression levels of BNIP3L,Parkinson disease protein 2(Parkin)and mitochondrial apopto-sis pathway-related proteins cleaved caspase-9 and cleaved caspase-3 were detected by Western blot.The co-localization of BNIP3L and the mitochondrial marker protein TOMM20 was detected by immunofluorescence staining.And,transmission electron microscopy was used to determine the number of mitophagosomes in neurons.Finally,apoptosis was detected by FITC Annexin V staining.The results showed that the expression of BNIP3L was significantly increased(P＜0.01),and the co-localization of BNIP3L and TOMM20 was increased after Cd treatment for 12 h.Silencing BNIP3L significantly inhibited Cd-induced Parkin translocation to mitochondria(P＜0.01),inhibited Cd-in-duced mitophagosome formation,and significantly promoted Cd-induced caspase-9 and caspase-3 activation and neuronal apoptosis(P＜0.01).The present results indicated that BNIP3L-mediated mitophagy was able to inhibit Cd-induced apoptosis in rat cerebral cortical neurons.
闻双全;王亮;方兰;袁燕
苏州健雄职业技术学院,江苏太仓 215411扬州大学兽医学院,江苏扬州 225009||江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州 225009
畜牧业
镉大鼠大脑皮质神经元BNIP3L线粒体自噬凋亡
cadmiumrat cerebral cortical neuronBNIP3Lmitophagyapoptosis
《畜牧与兽医》 2025 (001)
50-56 / 7
江苏省自然科学基金项目(BK20221372);江苏省高等学校基础科学研究面上项目(24KJB330009,22KJB360016);博士启动资金项目(2023);太仓基础研究项目(TC2022JC30);高等学校学科创新引智计划资助项目(D18007);江苏高校优势学科建设工程资助项目(PAPD)
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