Liu Bo, Gu Yan, Luo Jian-Hua, Fan Yuan-Shuo, Feng Zhi-Yu
Endocrinology department, Guizhou Provincial People's Hospital, Guiyang 550002.
Oncology department, Guizhou Provincial People's Hospital, Guiyang 550002, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2021 Sep;37(5):561-565. doi: 10.12047/j.cjap.6128.2021.072.
To investigate the effects of inhibition of lncRNA PVT1 on the proliferation, apoptosis and oxidative stress of vascular endothelial cells induced by hyperglycemic. Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and divided into four groups: control group (5.5 mmol/L glucose), high glucose group (30 mmol/L glucose), high glucose + siNC group (30 mmol/L glucose +siNC, negative control group), HG + siPVT1 group (30 mmol/L glucose + siPVT1, lncRNA PVT1 silencing group). The expression of PVT1 after transfection was detected by quantitative real-time PCR. MTT assay was used to detect the effect of siPVT1 (small interfering RNA PVT1) on the proliferation of HUVECs cells induced by high glucose. Flow cytometry was used to detect ROS and apoptosis of HUVECs cells induced by siPVT1. Western blot was used to detect the expression levels of apoptotic proteins such as Bax, Bcl-2, and cleaved caspase-3 in HUVECs cells. Compared with the control group, after transfection with siPVT1, the expression level of PVT1 was decreased significantly (<0.05). MTT results showed that the proliferation activity of HUVECs cells in the high-glucose group was reduced significantly after 24 h and 48 h. Compared with the HG + siNC group, the proliferation activity of HUVECs cells in the HG + siPVT1 group was increased significantly (<0.05) after 24 h and 48 h. Flow cytometry results showed that ROS and apoptosis rate of HUVECs cells in the high-glucose group were increased significantly compared with the control group. Compared with the HG + siNC (negative control) group, ROS and apoptosis rates of HUVECs cells in the HG + siPVT1 group were reduced significantly. Compared with the control group, the expression levels of cleaved-caspase-3 and Bax in the high-glucose group were significantly up-regulated, while the expression level of Bcl-2 was down-regulated. Compared with the HG + siNC group, the expression levels of cleaved-caspase-3 and Bax were down-regulated, and the expression level of Bcl-2 was up-regulated. The differences were statistically significant (<0.05). Inhibition of lncRNA PVT1 can significantly increase the proliferation activity of HUVECs cells induced by hyperglycemia, reduce oxidative stress and inhibit cell apoptosis.
探讨抑制lncRNA PVT1对高糖诱导的血管内皮细胞增殖、凋亡及氧化应激的影响。体外培养人脐静脉内皮细胞(HUVECs),分为四组:对照组(5.5 mmol/L葡萄糖)、高糖组(30 mmol/L葡萄糖)、高糖+siNC组(30 mmol/L葡萄糖+siNC,阴性对照组)、HG+siPVT1组(30 mmol/L葡萄糖+siPVT1,lncRNA PVT1沉默组)。采用定量实时PCR检测转染后PVT1的表达。采用MTT法检测siPVT1(小干扰RNA PVT1)对高糖诱导的HUVECs细胞增殖的影响。采用流式细胞术检测siPVT1诱导的HUVECs细胞的活性氧(ROS)和凋亡情况。采用蛋白质免疫印迹法检测HUVECs细胞中凋亡蛋白Bax、Bcl-2和裂解的caspase-3的表达水平。与对照组相比,转染siPVT1后,PVT1表达水平显著降低(<0.05)。MTT结果显示,高糖组HUVECs细胞在24 h和48 h后的增殖活性显著降低。与HG+siNC组相比,HG+siPVT1组HUVECs细胞在24 h和48 h后的增殖活性显著增加(<0.05)。流式细胞术结果显示,与对照组相比,高糖组HUVECs细胞的ROS水平和凋亡率显著增加。与HG+siNC(阴性对照)组相比,HG+siPVT1组HUVECs细胞的ROS水平和凋亡率显著降低。与对照组相比,高糖组裂解的caspase-3和Bax的表达水平显著上调,而Bcl-2的表达水平下调。与HG+siNC组相比,裂解的caspase-3和Bax的表达水平下调,Bcl-2的表达水平上调。差异具有统计学意义(<0.05)。抑制lncRNA PVT1可显著提高高糖诱导的HUVECs细胞的增殖活性,降低氧化应激并抑制细胞凋亡。