BNIP3介导线粒体自噬在m.3635G>A相关Leber遗传性视神经病变中的作用机制  

Mechanism of BNIP3-mediated mitophagy in m.3635G>A related Leber hereditary optic neuropathy

作　　者：刘贞 管威 张娟娟 管敏鑫 Liu Zhen;Guan Wei;Zhang Juanjuan;Guan Minxin(School of Laboratory Medicine and Life Sciences,Attardi Institute of Mitochondrial Biomedicine,Wenzhou Medical University,Wenzhou,Zhejiang 325035,China;State Key Laboratory of Ophthalmology,Optometry and Vision Science,Eye Hospital of Wenzhou Medical University,Wenzhou,Zhejiang 325027,China;Institute of Genetics,Zhejiang University,Zhejiang Provincial Key Lab of Genetic and Developmental Disorders,Hangzhou,Zhejiang 310058,China)

机构地区：[1]温州医科大学检验医学院、生命科学学院,Attardi线粒体生物医学研究院,温州325035 [2]温州医科大学附属眼视光医院省部共建眼视光学和视觉科学国家重点实验室,温州325027 [3]浙江大学遗传学研究所,浙江省遗传缺陷与发育障碍研究重点实验室,杭州310058

出　　处：《中华医学遗传学杂志》2025年第2期198-205,共8页Chinese Journal of Medical Genetics

基　　金：国家自然科学基金(82071007);浙江省自然科学基金(LY24C060001);温州市基础性医疗卫生科技项目(Y20220152)。

摘　　要：目的探讨BNIP3介导线粒体自噬在m.3635G>A变异相关Leber遗传性视神经病变(LHON)中的作用机制。方法选取2013年9月于温州医科大学附属眼视光医院就诊的1例携带m.3635G>A变异的中国LHON患者的转线粒体细胞系为研究对象,同时纳入1例具有相同线粒体背景的正常受试者的转线粒体细胞系作为对照,通过免疫荧光、实时荧光定量PCR(RT-qPCR)和蛋白免疫印迹等方法检测细胞中自噬相关蛋白的表达,探究BNIP3介导线粒体自噬对m.3635G>A变异相关LHON的作用。本研究已通过温州医科大学附属眼视光医院医学伦理委员会的审查(批准号:2023-J-096)。结果①与对照组细胞比较,变异组(m.3635G>A)细胞中自噬相关蛋白LC3(LC3-Ⅱ/LC3-Ⅰ)和LAMP1的蛋白表达水平均显著降低(P<0.05),其中大自噬相关蛋白ATG12、ATG7和ATG5的蛋白表达水平均显著降低(P<0.05)。②与对照组细胞比较,变异组细胞中线粒体自噬相关蛋白BNIP3的mRNA与蛋白表达水平均显著降低(P<0.05)。结论m.3635G>A变异可抑制BNIP3介导线粒体自噬的作用,进而导致LHON的发生。ObjectiveTo explore the mechanism of BNIP3-mediated mitophagy in m.3635G>A related Leber′s hereditary optic neuropathy(LHON).MethodsA transmitochondrial cybrid cell line derived from a Chinese LHON patient carrying the m.3635G>A,diagnosed at the Eye Hospital of Wenzhou Medical University in September 2013,was selected as the study subject.A transmitochondrial cybrid cell line from a healthy control with an identical mitochondrial background was included as a control.Immunofluorescence,real-time quantitative PCR(RT-qPCR),and Western blotting were employed to assess the expression of autophagy-related proteins,aiming to explore the role of BNIP3-mediated mitophagy in m.3635G>A related LHON.This study was approved by the Medical Ethics Committee of the Eye Hospital of Wenzhou Medical University(Ethics No.2023-J-096).Results①Compared with the control group,the protein expression levels of autophagy-related markers LC3(LC3-Ⅱ/LC3-Ⅰ)and LAMP1 were significantly reduced in the variant group(P<0.05).Additionally,the protein levels of macroautophagy-related proteins ATG12,ATG7,and ATG5 were also significantly decreased(P<0.05).②Compared with the control cells,the mRNA and protein expression levels of mitophagy-associated protein BNIP3 were significantly reduced in the cells of the variant group(P<0.05).Compared with the control group,both mRNA and protein expression levels of the mitophagy-related protein BNIP3 were significantly reduced in the variant group(P<0.05).ConclusionThe m.3635G>A inhibits BNIP3-mediated mitophagy,thereby contributing to the pathogenesis of LHON.

关 键 词：LEBER遗传性视神经病变 线粒体DNA 基因变异 自噬 线粒体自噬 

分 类 号：R774.1[医药卫生—眼科]