Department of Ophthalmology, Ning Xia Eye Hospital, People's Hospital of Ningxia Hui Autonomous Region (First Affiliated Hospital of Northwest University for Nationalities, Ningxia Clinical Research Center on Diseases of Blindness in Eye), Yinchuan, Ningxia, China.
Department of Pathology, School of Basic Medicine, Ningxia Medical University, Yinchuan, Ningxia, China.
PeerJ. 2022 May 31;10:e13375. doi: 10.7717/peerj.13375. eCollection 2022.
Herein, we aimed to present evidence that Ferulic acid (FA), a phenolic acid, can alleviate high glucose (HG)-induced retinal pigment epithelium (RPE) cell apoptosis and protect retina in db/db mice.
ARPE-19 cells (a human RPE cell line) were divided into four groups: control group; HG group (30 mmol/L glucose); HG+FA group (30 mmol/L glucose and 10 mmol/L FA). Cell viability and apoptosis were detected using CCK-8 and Annexin-5 staining, respectively. Apoptosis-related markers including P53, BAX and Bcl2 were examined by RT-qPCR, western blot and immunohistochemistry. Totally, 30 male db/db mice were randomly divided into db/db group (5 ml/kg saline) and FA group (0.05 g/kg FA). After treatment for 2 months, retinal samples were subjected to hematoxylin and eosin (H&E) and Masson staining. Moreover, immunofluorescence was used to detect apoptosis-related markers. Blood samples were collected for measuring cholesterol, triglyceride (TG), low-density lipoprotein (LDL) and high-density lipoprotein (HDL) levels.
FA treatment markedly increased cell viability and suppressed cell apoptosis of ARPE-19 cells compared to the HG-exposed group. Furthermore, FA ameliorated the abnormal expression levels of P53, BAX and Bcl2 in HG-induced ARPE-19 cells. In animal models, FA attenuated pathological changes in the retina tissues of diabetic mice. Consistent with models, FA significantly ameliorated the expression of apoptosis-related markers in retina tissues. Biochemical test results showed that FA reduced hyperlipidemia in diabetic mice.
Our findings suggest that FA alleviates HG-induced apoptosis in RPE cells and protects retina in db/db mice, which can be associated with P53 and BAX inactivation and Bcl2 activation.
本研究旨在提供证据表明,阿魏酸(FA)作为一种酚酸,可以减轻高葡萄糖(HG)诱导的视网膜色素上皮(RPE)细胞凋亡,并保护 db/db 小鼠的视网膜。
将 ARPE-19 细胞(人 RPE 细胞系)分为四组:对照组;HG 组(30mmol/L 葡萄糖);HG+FA 组(30mmol/L 葡萄糖和 10mmol/L FA)。分别用 CCK-8 和 Annexin-5 染色检测细胞活力和凋亡。通过 RT-qPCR、western blot 和免疫组化检测凋亡相关标志物,包括 P53、BAX 和 Bcl2。总共 30 只雄性 db/db 小鼠随机分为 db/db 组(5ml/kg 生理盐水)和 FA 组(0.05g/kg FA)。治疗 2 个月后,对视网膜样本进行苏木精和伊红(H&E)和 Masson 染色。此外,用免疫荧光检测凋亡相关标志物。收集血液样本以测量胆固醇、甘油三酯(TG)、低密度脂蛋白(LDL)和高密度脂蛋白(HDL)水平。
与暴露于 HG 的组相比,FA 处理显著增加了 ARPE-19 细胞的活力并抑制了细胞凋亡。此外,FA 改善了 HG 诱导的 ARPE-19 细胞中 P53、BAX 和 Bcl2 的异常表达水平。在动物模型中,FA 减轻了糖尿病小鼠视网膜组织的病变。与模型一致,FA 显著改善了视网膜组织中与凋亡相关的标志物的表达。生化测试结果表明,FA 降低了糖尿病小鼠的高脂血症。
我们的研究结果表明,FA 减轻了 HG 诱导的 RPE 细胞凋亡,并保护了 db/db 小鼠的视网膜,这可能与 P53 和 BAX 的失活以及 Bcl2 的激活有关。