苦参醇A对高糖诱导人视网膜血管内皮细胞增殖凋亡的影响研究  

Effect of Kushenol A on the Proliferation and Apoptosis of Human Retinal Vascular Endothelial Cell Induced by High Glucose

作　　者：王星力 柳维艳[1] 杨立宇 张勇[1] Wang Xingli;Liu Weiyan;Yang Liyu;Zhang Yong(Ophthalmology,Shenyang Fourth Peopled Hospital,Shenyang 110031,China)

机构地区：[1]沈阳市第四人民医院眼科,沈阳110031

出　　处：《中国药师》2021年第4期683-688,共6页China Pharmacist

摘　　要：目的:探讨苦参醇A(KA)对高糖诱导人视网膜血管内皮细胞增殖、凋亡的影响及其作用机制。方法:体外培养人视网膜血管内皮细胞,使用高糖诱导细胞,分别加入不同剂量的KA处理细胞;采用CCK-8检测细胞增殖;流式细胞术检测细胞凋亡率;采用qRT-PCR检测LncRNA DLX6-AS1、miR-145-5p的表达量;双荧光素酶报告实验与RIP实验验证DLX6-AS1、miR-145-5p的靶向关系;分别将si-DLX6-AS1、pc DNA-DLX6-AS1转染至高糖诱导的细胞中,采用上述方法检测细胞增殖及凋亡能力。结果:高糖诱导细胞可降低细胞活力、增高凋亡率(P<0.05),DLX6-AS1的表达水平显著升高(P<0.05),miR-145-5p的表达水平显著降低(P<0.05),而KA可明显逆转高糖对细胞增殖、凋亡及DLX6-AS1、miR-145-5p表达的作用(P<0.05),且呈剂量依赖性;双荧光素酶报告实验与RIP实验证实DLX6-AS1能够靶向结合miR-145-5p;转染si-DLX6-AS1后细胞活力升高(P<0.05),凋亡率降低(P<0.05),而转染pc DNA-DLX6-AS1能够明显减弱KA对高糖诱导视网膜血管内皮细胞增殖、凋亡的作用。结论:苦参醇A可能通过调控DLX6-AS1/miR-145-5p分子轴从而促进高糖诱导的人视网膜血管内皮细胞增殖及抑制细胞凋亡。Objective: To explore the effect of Kushenol A(KA) on the proliferation and apoptosis of human retinal vascular endothelial cell induced by high glucose and its mechanism. Methods: Human retinal vascular endothelial cells were cultured in vitro,high glucose induced cells were used,and different doses of KA treated cells were added respectively. CCK-8 was used to detect the cell proliferation. Flow cytometry was used to detect the apoptosis rate. The expressions of LncRNA DLX6-AS1 and miR-145-5p were detected by qRT-PCR. The dual luciferase report experiment and RIP experiment were used to verify the targeting relationship of DLX6-AS1 and miR-145-5p. si-DLX6-AS1 and pc DNA-DLX6-AS1 was transfected into the cells induced by high glucose,respectively,and the cell proliferation and apoptosis were detected by the above methods as well. Results: High glucose induction could reduce the cell viability(P<0.05),increase the apoptosis rate(P< 0.05),and the expression level of DLX6-AS1 was significantly increased(P <0.05),and the expression level of miR-145-5p was significantly reduced(P<0.05),while KA could significantly reverse the effects of high glucose on the cell proliferation and apoptosis,and the expression levels of DLX6-AS1 and miR-145-5p(P<0.05). The dual luciferase reporter experiment and the RIP experiment confirmed that DLX6-AS1 could target and bind to miR-145-5p. After transfection with si-DLX6-AS1,the cell viability was significantly increased(P< 0.05),and the apoptosis rate was significantly reduced(P< 0.05),while transfection with pcDNA-DLX6-AS1 could significantly weaken the effects of KA on the proliferation and apoptosis of retinal vascular endothelial cells induced by high glucose. Conclusion: Kushenol A may promote the proliferation of human retinal vascular endothelial cells induced by high glucose and inhibit the apoptosis by regulating the molecular axis of DLX6-AS1/miR-145-5p.

关 键 词：苦参醇A 高糖 人视网膜血管内皮细胞 增殖 凋亡 

分 类 号：R965.1[医药卫生—药理学]