Li Qian, Pang Lei, Shi Hui, Yang Wei, Liu Xin, Su Guanfang, Dong Yu
Department of Ophthalmology, The Second Hospital of Jilin University Changchun, Jilin, China.
Department of Ophthalmology, The First Hospital of Jilin University Changchun, Jilin, China.
Int J Clin Exp Pathol. 2018 May 1;11(5):2401-2407. eCollection 2018.
Diabetic retinopathy (DR) is a retinal disease in patients with diabetes caused by metabolic disorders of glucose, and often leads to irreversible blindness if not treated properly. Dysfunction of retinal endothelial cells (RECs) contributes to the pathogenesis of DR. In the present study we investigated the apoptotic effect of high glucose concentration. High glucose was used to induce cell injury. LDH assay kit was used to detect the leakage of LDH. Western blot was used to detect the expression of bax, cleaved-caspase-3, bcl-2 and p53 proteins. The transmission electron microscope was used to observe apoptotic morphological changes. In the present study, we found that the LDH leakage ratio was significant increased after exposing to 30 mmol/L glucose for 12 h, and both showed a concentration-department manner and time dependent fashion. Next, obvious apoptotic morphological changes were observed after treating the cells with glucose at 30 mmol/L. Further research indicated that the protein levels of p53, pro-apoptotic cleaved-caspase-3, and bax were significantly upregulated, and the level of bcl-2 was decreased after treating with glucose at 30 mmol/L. The protein levels of bax and cl-caspase-3 were significantly decreased and the expression of bcl-2 was increased after pretreated with p53 specific inhibitor pifithrin-α. In conclusion, high glucose concentration induces apoptosis in retinal endothelial cells by activating p53 signaling pathway.
糖尿病视网膜病变(DR)是糖尿病患者因葡萄糖代谢紊乱引起的一种视网膜疾病,若治疗不当常导致不可逆失明。视网膜内皮细胞(RECs)功能障碍在DR发病机制中起作用。在本研究中,我们调查了高糖浓度的凋亡效应。使用高糖诱导细胞损伤。采用乳酸脱氢酶(LDH)检测试剂盒检测LDH泄漏情况。用蛋白质免疫印迹法检测bax、裂解的半胱天冬酶-3、bcl-2和p53蛋白的表达。用透射电子显微镜观察凋亡形态学变化。在本研究中,我们发现暴露于30 mmol/L葡萄糖12小时后,LDH泄漏率显著增加,且呈浓度-时间依赖性。接下来,用30 mmol/L葡萄糖处理细胞后观察到明显的凋亡形态学变化。进一步研究表明,用30 mmol/L葡萄糖处理后,p53、促凋亡的裂解半胱天冬酶-3和bax的蛋白水平显著上调,bcl-2水平降低。用p53特异性抑制剂pifithrin-α预处理后,bax和cl-半胱天冬酶-3的蛋白水平显著降低,bcl-2表达增加。总之,高糖浓度通过激活p53信号通路诱导视网膜内皮细胞凋亡。