PDF(Pubmed)
Abstract：
BACKGROUND: Transient Receptor Potential Mucolipin 1 (TRPML1) serves as a pivotal reactive oxygen species (ROS) sensor in cells, which is implicated in the regulation of autophagy. However, its function in melanocyte autophagy under oxidative stress remains elusive.
METHODS: The expression and ion channel function of TRPML1 were investigated using immunofluorescence and calcium imaging in primary human melanocytes (MCs). After activating TRPML1 with MLSA1 (TRPML1 agonist), autophagy-related molecules were investigated via western blot. ROS level, apoptosis- and autophagy-related molecules were investigated after pretreatment with MLSA1. After interference with TRPML1 expression, mitochondrial structures were visualized by electron microscopy with hydrogen peroxide (H2O2）treatment.
RESULTS: TRPML1 was expressed and functionally active in primary human MCs, and its activation promotes elevated expression of LC3-II and reduced apoptosis and ROS levels under oxidative stress. TRPML1 downregulation caused mitochondrial swelling and disruption of cristae structures under oxidative stress in primary human MCs.
CONCLUSIONS: TRPML1 might mediate lysosomal autophagy in primary human MCs under oxidative stress, participating in mechanisms that maintain the oxidative and antioxidant systems in balance.
METHODS: The expression and ion channel function of TRPML1 were investigated using immunofluorescence and calcium imaging in primary human melanocytes (MCs). After activating TRPML1 with MLSA1 (TRPML1 agonist), autophagy-related molecules were investigated via western blot. ROS level, apoptosis- and autophagy-related molecules were investigated after pretreatment with MLSA1. After interference with TRPML1 expression, mitochondrial structures were visualized by electron microscopy with hydrogen peroxide (H2O2）treatment.
RESULTS: TRPML1 was expressed and functionally active in primary human MCs, and its activation promotes elevated expression of LC3-II and reduced apoptosis and ROS levels under oxidative stress. TRPML1 downregulation caused mitochondrial swelling and disruption of cristae structures under oxidative stress in primary human MCs.
CONCLUSIONS: TRPML1 might mediate lysosomal autophagy in primary human MCs under oxidative stress, participating in mechanisms that maintain the oxidative and antioxidant systems in balance.
摘要：
背景：瞬时受体电位黏磷脂1（TRPML1）在细胞中充当关键的活性氧（ROS）传感器，这与自噬的调节有关。然而,其在氧化应激下的黑素细胞自噬中的功能仍然难以捉摸。
方法：使用免疫荧光和钙成像研究了TRPML1在人原代黑素细胞（MCs）中的表达和离子通道功能。用MLSA1（TRPML1激动剂）激活TRPML1后，通过蛋白质印迹研究自噬相关分子。ROS水平,用MLSA1预处理后研究凋亡和自噬相关分子.干扰TRPML1表达后，用过氧化氢（H2O2）处理的电子显微镜观察线粒体结构。
结果：TRPML1在原代人MC中表达并具有功能活性，其激活促进LC3-II的表达升高，并减少氧化应激下的细胞凋亡和ROS水平。TRPML1下调导致原代人MC在氧化应激下线粒体肿胀和cr结构破坏。
结论：TRPML1可能在氧化应激下介导人原发性MC的溶酶体自噬，参与维持氧化和抗氧化系统平衡的机制。
方法：使用免疫荧光和钙成像研究了TRPML1在人原代黑素细胞（MCs）中的表达和离子通道功能。用MLSA1（TRPML1激动剂）激活TRPML1后，通过蛋白质印迹研究自噬相关分子。ROS水平,用MLSA1预处理后研究凋亡和自噬相关分子.干扰TRPML1表达后，用过氧化氢（H2O2）处理的电子显微镜观察线粒体结构。
结果：TRPML1在原代人MC中表达并具有功能活性，其激活促进LC3-II的表达升高，并减少氧化应激下的细胞凋亡和ROS水平。TRPML1下调导致原代人MC在氧化应激下线粒体肿胀和cr结构破坏。
结论：TRPML1可能在氧化应激下介导人原发性MC的溶酶体自噬，参与维持氧化和抗氧化系统平衡的机制。
