解放军医学杂志2025，Vol.50Issue(12)：1557-1566,10.DOI:10.11855/j.issn.0577-7402.1243.2025.0924
线粒体转录因子A对糖尿病心肌病小鼠心肌细胞氧化应激及其凋亡的影响
Effects of mitochondrial transcription factor A on oxidative stress and apoptosis in cardiomyocytes of mice with diabetic cardiomyopathy
摘要
Abstract
Objective To investigate the effects of mitochondrial transcription factor A(TFAM)in the myocardial tissue of mice with diabetic cardiomyopathy(DCM)on oxidative stress and apoptosis in cardiomyocytes.Methods Twenty-four male db/db mice were randomly divided into three groups(n=8 per group):db/db group,db/db+TFAM overexpression group and db/db+TFAM empty vector group.An additional 8 db/m male mice served as blank control group.Mice in db/db+TFAM overexpression group received a tail vein injection of MyoAAV1A-TFAM at a dose of 5×1011 vg/mouse to establish the TFAM overexpression model.db/db+TFAM empty vector group received a tail vein injection of an equivalent total amount of MyoAAV1A serotype-CON,while db/db group received an equal volume of normal saline via tail vein injection.The mRNA expression levels of TFAM and mitochondrial DNA(mtDNA)in myocardial tissue were detected by PCR.The reactive oxygen species(ROS)content in myocardial tissue was measured using the 20,70-dichlorodihydrofluorescein diacetate(DCFH-DA)probe.The activity of superoxide dismutase(SOD)and the contents of malondialdehyde(MDA)and ATP in myocardial tissue were assessed using commercial assay kits.The apoptosis rate of cardiomyocytes was determined by TUNEL staining.The protein expression levels of TFAM,Caspase-3,B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)in myocardial tissue were detected by Western blotting,and cardiac function indices of mice were measured using a small animal ultrasound system.Results Compared with db/m group,the expression levels of TFAM protein(P<0.01),mitochondrial ATP content(P<0.0001),and mtDNA expression level(P<0.001)in myocardial tissue of db/db mice were significantly reduced;the contents of ROS and MDA were significantly increased(P<0.0001),and the activity of SOD was decreased(P<0.001);the rate of TUNEL-positive cells in myocardial tissue increased(P<0.0001),the expression levels of Caspase-3 and Bax proteins were increased,and the expression level of Bcl-2 protein significantly reduced(P<0.0001).Compared with db/db group and the db/db+TFAM empty vector group,the mitochondrial ATP content and mtDNA expression level in myocardial tissue of mice in db/db+TFAM overexpression group were significantly increased(P<0.01),the contents of ROS and MDA were significantly decreased(P<0.0001),and the activity of SOD was increased(P<0.05);the rate of TUNEL-positive cells in myocardial tissue was significantly decreased(P<0.01);the expression levels of Caspase-3 and Bax proteins were decreased(P<0.0001),and the expression level of Bcl-2 protein was significantly increased(P<0.01).HE staining showed that compared with db/m group,myocardial cells in mice of db/db group were significantly hypertrophic and necrotic,and the arrangement and structure of myocardial cells were disordered;Compared with db/db group and db/db+TFAM empty vector group,the disorder of myocardial fibers in mice of db/db+TFAM overexpression group was improved,and the number of necrotic or inflammatory cells was significantly reduced.Cardiac color Doppler ultrasound showed that compared with db/m group,the left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),and E/A ratio in mice of db/db group were significantly decreased(P<0.0001);Compared with db/db group and db/db+TFAM empty vector group,the LVEF,LVFS and E/A ratio in mice of db/db+TFAM overexpression group were significantly increased(P<0.001).Conclusions TFAM protein may alleviate diabetes-induced myocardial damage by improving mitochondrial function,inhibiting oxidative stress,and suppressing apoptosis.关键词
线粒体转录因子A/细胞凋亡/氧化应激/心肌组织/糖尿病心肌病Key words
mitochondrial transcription factor A/cardiomyocyte apoptosis/oxidative stress/myocardial tissue/diabetic cardiomyopathy分类
医药卫生引用本文复制引用
Lyu Li-Li,Zhang Yue-Mei,Chen Yong-Qing,Wu Bing,Gu Hua,Song Xiao-Gang..线粒体转录因子A对糖尿病心肌病小鼠心肌细胞氧化应激及其凋亡的影响[J].解放军医学杂志,2025,50(12):1557-1566,10.基金项目
甘肃省自然科学基金(JRRA1377) （JRRA1377）
兰州市科技计划项目(2022-5-84) This work was supported by the Natural Science Foundation of Gansu Province(JRRA1377),and the Lanzhou Science and Technology Project(2022-5-84) （2022-5-84）
