LncRNA BRE-AS1在急性心肌梗死患者血清中表达及对缺氧/复氧诱导的心肌细胞损伤的影响

LncRNA BRE-AS1 expression in the serum of patients with acute myocardial infarction and its effect on cardiomyocyte injury induced by hypoxia/reoxygenation

目的探究lncRNA BRE-AS1在急性心肌梗死(AMI)患者血清中表达和对缺氧/复氧(H/R)诱导心肌细胞AC16损伤的影响。方法QPCR检测AMI患者血清和H/R处理12 h、24 h、48 h后AC16细胞中lncRNA BRE-AS1表达;将lncRNA BRE-AS1 siRNA和对照siRNA转染至AC16细胞中,qPCR检测检测转染后细胞中lncRNA BRE-AS1表达;利用试剂盒检测细胞中氧化应激指标MDA、SOD和GSH-PX含量,MTT检测细胞增殖,流式细胞术检测细胞凋亡,Western blot检测细胞中p38/MAPK信号通路蛋白表达。结果与健康志愿者和常氧组AC16细胞相比,AMI患者血清和H/R处理12 h、24 h、48 h后细胞中lncRNA BRE-AS1表达显著增加(P<0.01)。与沉默对照组比较,沉默BRE-AS1组AC16细胞中lncRNA BRE-AS1表达显著降低(P<0.01)。与常氧组比较,H/R组AC16细胞中MDA含量、细胞凋亡率、p38/MAPK信号通路蛋白p-p38、p-ERK1/2表达显著增加,SOD和GSH-PX含量及细胞增殖能力显著降低(P<0.01);与H/R+沉默对照组比较,H/R+沉默BRE-AS1组AC16细胞中MDA含量、细胞凋亡率、p38/MAPK信号通路蛋白p-p38、p-ERK1/2表达显著降低,SOD和GSH-PX含量及细胞增殖能力显著增加(P<0.01)。结论LncRNA BRE-AS1在AMI患者血清中高表达,沉默lncRNA BRE-AS1能够抑制H/R诱导的心肌细胞损伤,其机制可能与抑制p38/MAPK信号通路有关。

Objective To investigate the expression of lncRNA BRE-AS1 in the serum of patients with acute myocardial infarction(AMI)and the effect of hypoxia/reoxygenation(H/R)-induced cardiomyocytes AC16 injury.Methods QPCR was used to detect the expression of lncRNA BRE-AS1 in AC16 cells after 12 h,24 h,and 48 h of AMI patients'serum and H/R treatment.LncRNA BRE-AS1 siRNA and control siRNA were transfected into AC16 cells,and qPCR detection detected the expression of lncRNA BRE-AS1 in the transfected cells.The content of oxidative stress indicators MDA,SOD and GSH-PX in the cells was detected by using the kit,MTT was used to detect cell proliferation,flow cytometry was used to detect apoptosis,and Western blot was used to detect the expression of p38/MAPK signal pathway protein in cells.Results Compared with healthy volunteers and AC16 cells in the normoxic group,the expression of lncRNA BRE-AS1 in cells of AMI patients was significantly increased after treatment with H/R for 12 h,24 h,and 48 h(P<0.01).Compared with the silent control group,the expression of lncRNA BRE-AS1 in AC16 cells of the silent BRE-AS1 group was significantly reduced(P<0.01).Compared with the normoxic group,the MDA content,apoptosis rate,p38/MAPK signaling pathway protein p-p38 and p-ERK1/2 expression in AC16 cells in the H/R group were significantly increased,and the content of SOD and GSH-PX and cell proliferation ability Significantly reduced(P<0.01).Compared with the H/R+silent control group,the MDA content,apoptosis rate,p38/MAPK signaling pathway protein p-p38,p-ERK1/2 expression in AC16 cells in the H/R+silent BRE-AS1 group were significantly reduced,SOD and GSH-PX content and cell proliferation capacity increased significantly(P<0.01).Conclusion LncRNA BRE-AS1 is highly expressed in the serum of patients with AMI.Silencing lncRNA BRE-AS1 can inhibit H/R-induced myocardial cell injury.The mechanism may be related to the inhibition of p38/MAPK signaling pathway.