摘要
目的 探究长链非编码RNA(lncRNA)浆细胞瘤转化迁移基因1(PVT1)靶向调控miR-625-5p对高糖诱导心肌细胞损伤的影响及其机制。方法 将心肌H9c2细胞分为NG组、HG组分别用5.5、30.0 mmol/L葡萄糖干预,再将30.0 mmol/L葡萄糖干预的心肌H9c2细胞分别转染si-NC、si-PVT1、miR-NC、miR-625-5p、si-PVT1+anti-miR-NC、si-PVT1+anti-miR-625-5p,记为HG+si-NC组、HG+si-PVT1组、HG+miR-NC组、HG+miR-625-5p组、HG+si-PVT1+anti-miR-NC组、HG+si-PVT1+anti-miR-625-5p组。qRT-PCR检测PVT1、miR-625-5p表达量;流式细胞术检测细胞凋亡情况;Western blot检测Cleaved-caspase3、Bax蛋白表达;ELISA法检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)表达水平;双荧光素酶报告实验验证PVT1与miR-625-5p的靶向关系。结果 与NG组比较,HG组PVT1表达量、凋亡率、Cleaved-caspase3蛋白、Bax蛋白、TNF-α、IL-1β、IL-6表达水平升高,miR-625-5p表达量降低(P<0.05)。与HG+si-NC组比较,HG+si-PVT1组PVT1表达量、凋亡率、Cleaved-caspase3蛋白、Bax蛋白、TNF-α、IL-1β、IL-6表达水平降低,miR-625-5p表达量升高(P<0.05)。与HG+miR-NC组比较,HG+miR-625-5p组miR-625-5p表达量升高,Cleaved-caspase3蛋白、Bax蛋白、TNF-α、IL-1β、IL-6表达水平降低(P<0.05)。与HG+si-PVT1+anti-miR-NC组比较,HG+si-PVT1+anti-miR-625-5p组miR-625-5p表达量降低,凋亡率、Cleaved-caspase3蛋白、Bax蛋白、TNF-α、IL-1β、IL-6表达水平升高(P<0.05)。结论 PVT1可通过靶向负调控miR-625-5p减轻高糖诱导的心肌细胞凋亡和炎症反应损伤。
Objective To explore the effect and mechanism of long non-coding RNA(lncRNA)Plasmacytoma varant translocation 1(PVT1)targeting miR-625-5 p on high glucose-induced cardiomyocyte injury.Methods Myocardial H9 c2 cells were divided into NG group and HG group,which were intervened with 5.5 and 30.0 mmol/L glucose,respectively,and then the myocardial H9 c2 cells intervened with 30.0 mmol/L glucose were transfected with si-NC,si-PVT1,miR-NC,and miR-625-5 p,si-PVT1+anti-miR-NC,and si-PVT1+anti-miR-625-5 p,respectively,denoted as HG+si-NC group,HG+si-PVT1 group,HG+miR-NC group,HG+miR-625-5 p group,HG+si-PVT1+anti-miR-NC group,HG+si-PVT1+anti-miR-625-5 p group.qRT-PCR was used to detect the expressions of PVT1 and miR-625-5 p,and flow cytometry was used to detect cell apoptosis.Western blot was used to detect the expressions of Cleaved-caspase3 and Bax,and ELISA was used to detect tumor necrosis factor-α(TNF-α),and the expression levels of interleukin-1β(IL-1β)and interleukin-6(IL-6).The dual-luciferase reporter assay verified the targeting relationship between PVT1 and miR-625-5 p.Results Compared with NG group,the expression of PVT1,apoptosis rate,and Cleaved-caspase3 protein,Bax protein,TNF-α,IL-1β,and IL-6 expression levels in HG group increased,while the expression of miR-625-5 p decreased(P<0.05).Compared with the HG+si-NC group,the PVT1 expression,apoptosis rate,Cleaved-caspase3 protein,Bax protein,TNF-α,IL-1β,and IL-6 expression levels in the HG+si-PVT1 group decreased,and the expression level of miR-625-5 p increased(P<0.05).Compared with the HG+miR-NC group,the expression of miR-625-5 p in the HG+miR-625-5 p group increased,and the expression levels of Cleaved-caspase3 protein,Bax protein,TNF-α,IL-1β,and IL-6 decreased(P<0.05).Compared with the HG+si-PVT1+anti-miR-NC group,the expression of miR-625-5 p in the HG+si-PVT1+anti-miR-625-5 p group decreased,while the apoptosis rate,the expression levels of Cleaved-caspase3 protein,Bax protein,TNF-α,IL-1βand IL-6 increased(P<0.05).Conclusion PVT1 attenuates
作者
黄岚
张洪波
庄红
HUANG Lan;ZHANG Hong-bo;ZHUANG Hong(Department of Laboratory,the Affiliated Hospital of Jianghan University the Sixth Hospital of Wuhan City,Wuhan 430015,China;Department of Cardiology,the Affiliated Hospital of Jianghan University the Sixth Hospital of Wuhan City,Wuhan 430015,China)
出处
《临床误诊误治》
CAS
2022年第8期105-111,共7页
Clinical Misdiagnosis & Mistherapy
基金
武汉市医学科研项目(WX19C39)。
