C-sis对缺氧导致心肌细胞凋亡的抑制作用  

Inhibitory Effect of C-sis on Cardiomyocyte Apoptosis

作　　者：阮起超 孙国芳[2] RUAN Qi-chao;SUN Guo-fang(Department of Gastroenterology,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,China;ECG Diagnostic Room,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第二附属医院消化内科,南昌330006 [2]南昌大学第二附属医院心电诊断室,南昌330006

出　　处：《南昌大学学报（医学版）》2023年第1期9-13,44,共6页Journal of Nanchang University：Medical Sciences

基　　金：江西省自然科学基金(20202BABL206010)。

摘　　要：目的探讨C-sis对缺氧诱导心肌细胞凋亡的作用。方法制备原代小鼠心肌细胞;将心肌细胞分为空白组、缺氧组、缺氧及空载质粒转染组和缺氧及pcDNA3.1/C-sis转染组;将空载质粒及pcDNA3.1/C-sis分别转染缺氧及空载质粒转染组和缺氧及pcDNA3.1/C-sis转染组心肌细胞;对缺氧组、缺氧及空载质粒转染组和缺氧及pcDNA3.1/C-sis转染组心肌细胞进行缺氧处理;倒置显微镜观察各组心肌细胞的搏动频率、异常搏动次数;全自动生化分析仪测定培养液中乳酸脱氢酶(LDH)含量,采用MTT法检测细胞存活率,流式细胞仪检测细胞凋亡率,蛋白质印迹法检测凋亡相关蛋白、C-sis蛋白表达水平。结果与空白组相比,缺氧组心肌细胞搏动频率明显变慢[(99±5)次·min^(-1)比(159±21)次·min^(-1),P<0.01],异常搏动次数明显增多[(16.9±1.4)次·min^(-1)比(2.4±0.7)次·min^(-1),P<0.01],细胞培养液中LDH含量明显升高[(88.27±13.14)U·L^(-1)比(23.54±5.81)U·L^(-1),P<0.01],细胞存活率降低[(57.54±4.84)%比(100.00±0.00)%,P<0.01],凋亡率升高[(39.26±4.05)%比(1.77±0.87)%,P<0.01],Caspase3和Bax蛋白表达明显升高[Caspase3:(2.27±0.94)比(0.22±0.11),P<0.01;Bax:(1.79±0.77)比(0.87±0.27),P<0.05],Bcl-2蛋白表达明显下降[(0.12±0.06)比(0.84±0.24),P<0.01]。而pcDNA3.1/C-sis质粒转染后C-sis表达明显升高,并抑制上述缺氧所致心肌细胞发生的一系列变化(P<0.01或P<0.05)。结论C-sis能够抑制缺氧所致的心肌细胞凋亡。Objective To investigate the effect of C-sis on cardiomyocyte apoptosis induced by hypoxia.Methods Primary mouse cardiomyocytes were prepared,and divided into blank group,hypoxia group,hypoxia+empty vector transfection group,and hypoxia+pcDNA3.1/C-sis transfection group.The hypoxia+empty vector transfection group and the hypoxia+pcDNA3.1/C-sis transfection group were transfected with empty plasmids and pcDNA3.1/C-sis,respectively.The hypoxia group and the hypoxia+empty vector transfection group were treated with hypoxia.The beat frequency and the number of abnormal beats of cardiomyocytes were observed under an inverted microscope.The content of lactate dehydrogenase(LDH)in the culture medium was measured using an automatic biochemical analyzer.Cell survival and apoptosis were determined by MTT assay and flow cytometry,respectively.The expression levels of apoptosis-related proteins and C-sis protein were detected by Western blotting.Results Compared with the blank group,the hypoxia group showed a significant decrease in beat frequency((99±5)beats·min-1 vs(159±21)beats·min-1),survival rate((57.54±4.84)%vs(100.00±0.00)%)and Bcl-2 expression(0.12±0.06 vs 0.84±0.24),as well as an increase in the number of abnormal beats((16.9±1.4)beats·min-1 vs(2.4±0.7)beats/min),content of LDH in culture medium((88.27±13.14)U·L^(-1) vs(23.54±5.81)U·L^(-1)),apoptosis rate(39.26±4.05)%vs(1.77±0.87)%),Caspase-3 expression((2.27±0.94)vs(0.22±0.11))and Bax expression((1.79±0.77)vs(0.87±0.27))(P<0.01 or P<0.05).However,pcDNA3.1/C-sis transfection increased C-sis expression,and inhibited hypoxia-caused changes in cardiomyocytes(P<0.01 or P<0.05).Conclusion C-sis can inhibit cardiomyocyte apoptosis induced by hypoxia.

关 键 词：C-SIS 缺氧 心肌细胞 细胞凋亡 动物 实验 小鼠 

分 类 号：R-332[医药卫生]