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胡椒碱通过调控H19/miR-29b通路改善高糖低氧诱导的视网膜色素上皮细胞损伤
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Abstract:
目的：研究胡椒碱对视网膜色素上皮(retinal pigment epithelium, RPE)细胞在高糖 + 低氧条件下的保护作用机制。方法：高糖(25 mM)及低氧(5% O2持续24小时)构建RPE细胞体外(D407细胞系)糖尿病视网膜病变模型。实验主要分为对照组、高糖低氧模型组及胡椒碱干预组等。通过RT-qPCR及免疫荧光法检测lncRNA H19、miR-29b及VEGFA的mRNA或蛋白表达水平，使用划痕实验评估D407细胞的迁移能力。结果：与对照组相比，高糖及低氧条件下，D407细胞lncRNA H19及VEGFA mRNA的转录增加，miR-29b的转录减少；VEGFA蛋白表达增加；细胞迁移能力下降。使用胡椒碱干预后，可部分逆转上述现象。结论：胡椒碱可能通过lncRNA H19/miR-29b通路减轻高糖低氧诱导的RPE细胞损伤，为糖尿病视网膜病变的防治提供了新的潜在治疗策略。
Objective: To investigate the protective mechanism of piperine on retinal pigment epithelium (RPE) cells under conditions of high glucose and hypoxia. Methods: An in vitro model of diabetic retinopathy was established using the D407 cell line, subjected to high glucose (25 mM) and hypoxic conditions (5% O？ for 24 hours). The experiments were primarily divided into a control group, a high glucose and hypoxia model group, and a piperine intervention group. The expression levels of long non-coding RNA (lncRNA) H19, microRNA-29b (miR-29b), and vascular endothelial growth factor A (VEGFA) mRNA were measured using reverse transcription quantitative polymerase chain reaction (RT-qPCR). VEGFA protein expression was assessed using immunofluorescence techniques. The migration ability of D407 cells was evaluated through a scratch assay. Results: Compared to the control group, the high glucose and low oxygen group exhibited increased expression levels of lncRNA H19 and VEGFA mRNA, decreased expression of miR-29b, elevated VEGFA protein expression, and reduced cell migration. Treatment with piperine partially reversed these effects. Conclusion: Piperine may mitigate high glucose and hypoxia-induced damage in RPE cells via the lncRNA H19/miR-29b pathway, offering a novel potential therapeutic strategy for the prevention and treatment of diabetic retinopathy.
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