山东医药

目的 探讨鞣花酸（EA）是否通过调控PTEN诱导假定激酶1(PINK1)/帕金森蛋白（Parkin）信号通路对高糖环境内皮细胞线粒体自噬产生影响。方法 取对数生长期人脐静脉融合细胞EA-hy926，随机分为对照组、模型组、EA组，每组3个复孔。模型组、EA组用含50 mmol/L葡萄糖的DMEM培养基培养24 h建立高糖损伤内皮细胞模型，EA组加入40 nmol/L EA工作液继续培养24 h，对照组不做任何处理。培养24 h时采用CCK-8法检测各组细胞增殖活性，采用成管实验观察细胞成管能力，透射电镜下观察线粒体结构，采用Western blotting法检测PINK1、Parkin蛋白，采用分析对接法分析EA与PINK1结合情况。结果 与对照组相比，模型组细胞存活率低、管腔总长度短（P均<0.05）；与模型组相比，EA组细胞存活率高、管腔总长度长（P均<0.05）。与对照组相比，模型组线粒体出现肿胀、嵴结构消失、空泡化等；与模型组相比，EA组大部分线粒体的受损嵴结构修复，线粒体空泡化改善，线粒体结构趋于正常。与对照组相比，模型组PINK1、Parkin蛋白相对表达量均低（P均<0.05）；与模型组相比，EA组PINK1、Parkin蛋白相对表达量均高（P均<0.05）。EA与PINK1的结合能为-11.5 kcal/mol,EA和PINK1蛋白的氨基酸残基形成6个氢键。结论 EA可能通过上调PINK1/Parkin信号通路，促进高糖环境下EA-hy926细胞的线粒体自噬，改善细胞的增殖活性和成管能力。

Objective To explore whether ellagic acid(EA) affects mitophagy in endothelial cells under high glucose environment by regulating the PTEN-induced kinase 1(PINK1)/Parkin signaling pathway. Methods EA-hy926 cells(human umbilical vein endothelial cells) in the logarithmic growth phase were randomly divided into the control group, model group, and EA group, with 3 replicate wells in each group. The model group and EA group were cultured in DMEM medium containing 50 mmol/L glucose for 24 h to establish the high glucose-induced endothelial cell injury models. The EA group was further cultured for 24 h after adding 40 nmol/L EA working solution, while the control group received no treatment. At 24 h of culture, CCK-8 assay was used to detect cell proliferation activity, tube formation assay was performed to observe cell tube formation ability, transmission electron microscopy was used to observe mitochondrial structure, Western blotting was applied to detect PINK1 and Parkin proteins, and molecular docking analysis was used to evaluate the binding between EA and PINK1. Results Compared with the control group, the model group showed lower cell survival rate and shorter total lumen length(both P<0. 05). Compared with the model group, the EA group had higher cell survival rate and longer total lumen length(both P<0. 05). Compared with the control group, the model group exhibited mitochondrial swelling, cristae loss, and vacuolization. In contrast, most mitochondria in the EA group showed repaired damaged cristae, alleviated vacuolization, and a near-normal morphological structure. The relative expression levels of PINK1 and Parkin proteins in the model group were significantly lower than those in the control group(both P<0. 05), while those in the EA group were significantly higher than those in the model group(both P<0. 05). The binding energy between EA and PINK1 was-11. 5 kcal/mol, and EA formed 6 hydrogen bonds with amino acid residues of PINK1 protein. Conclusion EA may promote mitophagy in EA-hy926 cells under high glucose environment by up-regulating the PINK1/Parkin signaling pathway, thereby improving cell proliferation activity and tube formation ability.