心脑血管病防治2023，Vol.23Issue(11)：11-17,7.DOI:10.3969/j.issn.1009-816x.2023.11.003
LncRNA GAS5通过miR-137/SIRT6轴抑制糖尿病大鼠心肌细胞凋亡的机制研究
Mechanism of long non-coding RNA growth arrest-specific transcripts 5 inhibiting the apoptosis of cardiomyocytes in diabetic rats via miR-137/silent information regulator 6 axis
摘要
Abstract
Objective To investigate the effect of long non-coding RNA growth arrest-specific transcripts 5(LncRNA GAS5)on the apoptosis of cardiomyocytes and its molecular mechanism in diabetic cardiomyopathy(DCM)rats.Methods DCM rat models were constructed by high-glucose high-fat diet combined with streptozotocin intraperitoneal injection,and were devided into Sham group and DCM group.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of myocardial tissues;Masson staining was used to detect the degree of myocardial fibrosis in rats.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of LncRNA GAS5 in myocardial tissues.Rat cardiomyocytes(H9c2)were randomly divided into the control group,the high glucose group,the high glucose+GAS5 group,the high glucose+miR-137 mimics group,the high glucose+miR-137 mimics+GAS5 group,the high glucose+SIRT6 group,and the high glucose+SIRT6+miR-137 mimics group for corresponding treatment.QRT-PCR detected the expression levels of LncRNA GAS5 and miR-137.Western blot detected the protein expression levels of B-cell lymphoma-2(Bcl-2),Bcl-2 associated x(Bax)and silent information regulator 6(SIRT6).Flow cytometry detected apoptosis.Cell counting kit 8 detected cell viability.Dual luciferase reporter gene assay verified the targeting relationship between LncRNA GAS5 and miR-137,miR-137 and SIRT6.Results HE staining and Masson staining results showed that compared with the Sham group,cardiomyocytes in the DCM group were hypertrophic,disorganized,with increased collagen fibers and a large number of inflammatory cells infiltrated.Compared with the Sham group,LncRNA GAS5 expression decreased in the DCM group(P<0.01).Compared with the control group,the expression level of LncRNA GAS5 in the high glucose group gradually decreased with the prolongation of high glucose incubation time(P<0.01).Compared with the high glucose group,LncRNA GAS5 expression,cell viability,and Bcl-2 expression increased in the high glucose+GAS5 group(P<0.05),while miR-137,cell apoptosis ratio,and Bax expression decreased(P<0.05).Compared with the high glucose group,miR-137,Bax protein expression,and cell apoptosis ratio increased in the high glucose+miR-137 mimics group,while cell viability,SIRT6,and Bcl-2 protein expressions decreased(P<0.05).Compared with the high glucose+miR-137 mimics group,cell viability,Bcl-2 expression increased in the high glucose+GAS5+miR-137 mimics group,while cell apoptosis ratio and Bax expression decreased(P<0.05).Compared with the high glucose group,cell viability,Bcl-2,and SIRT6 protein expressions increased in the high glucose+SIRT6 group(P<0.01),while cell apoptosis ratio and Bax expression decreased(P<0.05).Compared with the high glucose+SIRT6 group,cell apoptosis ratio and Bax protein expression increased in the high glucose+SIRT6+miR-137 mimics group(P<0.01),but cell viability and Bcl-2 protein expression decreased(P<0.01).Dual luciferase reporter gene assay confirmed the targeting relationship between LncRNA GAS5 and miR-137,miR-137 and SIRT6.Conclusion Overexpression of LncRNA GAS5 reduces the binding of miR-137 and SIRT6 through an endogenous competitive mechanism,and promotes SIRT6 expression to inhibit cardiomyocytes apoptosis in DCM rat.关键词
糖尿病心肌病/长链非编码RNA生长停滞特异性转录因子5/miR-137/沉默调节蛋白6/凋亡Key words
Diabetic cardiomyopathy/Long non-coding RNA growth arrest-specific transcripts 5/MiR-137/Silent information regulator 6/Apoptosis引用本文复制引用
王庆军,王天宇,胡鹏飞..LncRNA GAS5通过miR-137/SIRT6轴抑制糖尿病大鼠心肌细胞凋亡的机制研究[J].心脑血管病防治,2023,23(11):11-17,7.基金项目
浙江省医药卫生科技计划项目(2020KY205) （2020KY205）
浙江省中医药科技计划项目(2022ZA082) （2022ZA082）
浙江中医药大学中青年科研创新基金项目(KC201938) （KC201938）
