Astragalus Polysaccharide Regulates miR-182/Bcl-2 Axis to Relieve Metabolic Memory through Suppressing Mitochondrial Damage-Mediated Apoptosis in Retinal Pigment Epithelial Cells
Astragalus Polysaccharide Regulates miR-182/Bcl-2 Axis to Relieve Metabolic Memory through Suppressing Mitochondrial Damage-Mediated Apoptosis in Retinal Pigment Epithelial Cells
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DOI：
10.1159/000515901
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发表时间：
2021-08-05
期刊：
影响因子：
3.1
通讯作者：
Li, Wen-Jie
中科院分区：
文献类型：
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作者：
Gao, Li-Mo;Fu, Shun;Li, Wen-Jie
简介：代谢记忆是糖尿病视网膜病变的原因之一，黄芪多糖在治疗糖尿病方面具有很大的优势。然而，APS对代谢记忆的影响仍有待研究。研究方法：以视网膜色素上皮细胞株ARPE-19和原代视网膜色素上皮细胞为实验材料，观察黄芪多糖对高糖诱导的代谢记忆诱导的线粒体损伤和凋亡的影响。通过荧光素酶活性测定证实了miR-182和Bcl-2之间的关系。Western印迹和定量逆转录聚合酶链反应进行了调查线粒体损伤和凋亡相关标志物的变化。JC-1荧光法测定细胞线粒体膜电位。末端脱氧核苷酸转移酶dUTP缺口末端标记染色和流式细胞术分析，以确定凋亡的发生。结果如下：高糖处理后正常葡萄糖处理显著上调miR-182的表达，下调其靶基因Bcl-2的表达，APS处理可逆转上述效应。此外，APS治疗恢复线粒体功能，抑制细胞凋亡的代谢记忆状态。APS对miR-182过表达后线粒体损伤和细胞凋亡的影响被部分抑制。结论：黄芪多糖通过调节miR-182/Bcl-2轴减轻代谢记忆诱导的线粒体损伤和细胞凋亡，有望成为糖尿病视网膜病变治疗的新策略。
Introduction: Metabolic memory is one of the causes of diabetic retinopathy, and astragalus polysaccharide (APS) has great advantages in the treatment of diabetes. However, the effect of APS on metabolic memory remains to be investigated. Methods: Retinal pigment epithelial cell line ARPE-19 and primary retinal pigment epithelial cells were used to verify the effect of APS on mitochondria damage and apoptosis induced by high glucose-induced metabolic memory. The relationship between miR-182 and Bcl-2 was confirmed by a luciferase activity assay. Western blotting and quantitative reverse-transcriptase polymerase chain reaction were conducted to investigate the changes in mitochondrial damage- and apoptosis-associated markers. The cell mitochondrial membrane potential was assessed by JC-1 fluorescence. Terminal deoxynucleotidyl transferase dUTP nick end labelling staining and flow cytometry assays were performed to determine the occurrence of apoptosis. Results: Treatment with high glucose followed by normal glucose significantly upregulated the expression of miR-182 and downregulated the expression of its target Bcl-2, and APS treatment reversed the above effects. Additionally, APS treatment restored mitochondrial function and inhibited apoptosis in cells in a state of metabolic memory. The effects of APS against mitochondrial damage and apoptosis were partially inhibited after miR-182 overexpression. Conclusion: APS alleviated mitochondrial damage and apoptosis induced by metabolic memory by regulating the miR-182/Bcl-2 axis, which might serve as a new strategy for the treatment of diabetic retinopathy.