Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
Physiol Res. 2024 Apr 30;73(2):253-263. doi: 10.33549/physiolres.934925.
Up to now, there's a limited number of studies on the relationship between PINK1/Park2 pathway and mitophagy in NAFLD. To investigate the effect of Park2-mediated mitophagy on non-alcoholic fatty liver disease (NAFLD). Oleic acid was used for the establishment of NAFLD model. Oil red-dyed lipid drops and mitochondrial alternations were observed by transmission electron microscopy. Enzymatic kit was used to test lipid content. The levels of IL-8 and TNF-alpha were determined by ELISA. Lenti-Park2 and Park2-siRNA were designed to upregulate and downregulate Park2 expression, respectively. The changing expression of PINK and Park2 was detected by RT-qPCR and Western blot. Immunofluorescence staining was applied to measure the amount of LC3. Successful NAFLD modeling was featured by enhanced lipid accumulation, as well as the elevated total cholesterol (TC), triglyceride (TG), TNF-alpha and IL-8 levels. Mitochondria in NAFLD model were morphologically and functionally damaged. Park2 expression was upregulated by lenti-Park2 and downregulated through Park2-siRNA. The PINK1 expression showed the same trend as Park2 expression. Immunofluorescence staining demonstrated that the when Park2 was overexpressed, more LC3 protein on mitochondrial autophagosome membrane was detected, whereas Park2 knockdown impeded LC3' locating on the membrane. The transmission electron microscopy image exhibited that the extent of damage to the mitochondrial in NAFLD model was revered by enhanced Park2 expression but further exacerbated by reduced Park2 expression. Park2-mediated mitophagy could relive NAFLD and may be a novel therapeutic target for NAFLD treatment. Keywords: Non-alcoholic Fatty Liver Disease (NAFLD), Mitophagy, PINK1/Park2, Park2, PINK1.
到目前为止，关于 PINK1/Park2 通路与 NAFLD 中自噬的关系的研究数量有限。为了研究 Park2 介导的线粒体自噬对非酒精性脂肪性肝病（NAFLD）的影响。使用油酸建立了 NAFLD 模型。通过透射电子显微镜观察油红染色的脂质滴和线粒体变化。酶试剂盒用于测试脂质含量。ELISA 法测定 IL-8 和 TNF-α水平。设计了 Lenti-Park2 和 Park2-siRNA 分别上调和下调 Park2 表达。通过 RT-qPCR 和 Western blot 检测 PINK 和 Park2 的变化表达。免疫荧光染色用于测量 LC3 的量。成功建立的 NAFLD 模型的特征是脂质积累增加，总胆固醇（TC）、甘油三酯（TG）、TNF-α和 IL-8 水平升高。NAFLD 模型中的线粒体形态和功能受损。Lenti-Park2 上调 Park2 表达，Park2-siRNA 下调 Park2 表达。PINK1 表达与 Park2 表达呈相同趋势。免疫荧光染色表明，当 Park2 过表达时，在线粒体自噬体膜上检测到更多的 LC3 蛋白，而 Park2 敲低则阻止 LC3 定位在膜上。透射电子显微镜图像显示，通过增强 Park2 表达，NAFLD 模型中线粒体损伤的程度得到逆转，但通过降低 Park2 表达，损伤程度进一步加重。Park2 介导的线粒体自噬可以缓解 NAFLD，可能成为治疗 NAFLD 的新靶点。关键词：非酒精性脂肪性肝病（NAFLD）、线粒体自噬、PINK1/Park2、Park2、PINK1。
