摘要
目的探讨利拉鲁肽(LIRA)对高糖诱导的H9c2细胞凋亡的影响。方法在H9c2细胞的高糖培养液中加入LIRA、活性氧簇分子(ROS)清除剂N-乙酰半胱氨酸(NAC)、p38丝裂原活化蛋白激酶(p38MARK)抑制剂SB203580和c-Jun氨基末端激酶(JNK)抑制剂SP600125等不同的处理因素作用24h,采用流式细胞术检测细胞的凋亡,采用Westernblot法检测H9c2细胞活化的含半胱氨酸的天冬氨酸蛋白水解酶3(c-caspase-3)、丝裂原活化蛋白激酶(MAPK)家族磷酸化的p38MAPK(p-p38MAPK)和JNK(p-JNK)的蛋白水平。结果正常糖对照组、等渗透压对照组和高糖组(HG组)的细胞凋亡率分别为(1.04-I-0.11)%、(1.27±0.04)%和(28.60±1.08)%。与正常糖对照组比较,等渗透压对照组的细胞凋亡率差异无统计学意义(P〉0.05),而HG组的细胞凋亡率显著增加(P〈0.01)。HG+LIRA组、HG+NAC组、HG+SB203580组和HG+SP600125组的细胞凋亡率分别为(6.67±0.77)%、(3.65±0.26)%、(4.59±0.48)%和(4.22±0.89)%。与HG组比较,HG+LIRA组、HG+NAC组、HG+SB203580组和HG+sP600125组的凋亡率均明显下降(P〈0.01)。与正常糖对照组比较,HG组心肌细胞内c-caspase-3、p-p38MAPK和p-JNK的蛋白表达水平明显上升(P〈0.01)。与HG组比较,利拉鲁肽明显抑制了c-caspase-3蛋白水平(P〈0.01),降低了p-p38MAPK和P-JNK的蛋白表达水平(P〈0.05)。结论利拉鲁肽可能通过抑制p38MAPK和JNK的磷酸化抑制高糖培养的H9c2细胞凋亡。
Objective To investigate the effects of liraglutide on high glucose induced apoptosis of H9c2 cardiomyocyte. Methods Different treatment factors were added into the H9c2 cardiomyocyte culture solution for 24 h including liraglutide ( LIRA ), N-acetylcysteine ( NAC ) which is a reactive oxygenspecies(ROS) scavenger,SB203580 which is an inhibitor of p38 mitogen activated protein kinase (p38MARK) and SP600125 which is an inhibitor of c-Jun N-terminal kinase (JNK). Cell apoptosis was detected by flow cytometrywhile, Western blotting was used to detect the activation of cysteinyl aspartate specific proteinase-3 ( c-caspase-3 ), the phosphorylation levels of p38MAPK (p-p38MAPK) and JNK (p-JNK). Results The apoptosis rates of normal glucosecontrol group (control group), equal osmotic pressure group and high glucose group ( HG group ) were ( 1.04 ± 0. 11 )%, ( 1.27 ± 0. 04 )% and (28.60 ±1.08)% respectively. The apoptosis rates of HG + LIRA group,HG + NAC group,HG + SB203580 group and HG +SP600125 group were (6.67±0.77)%,(3.65±0.26)%,(4.59±0.48)% and (4.22±0.89)% respectively. Compared with the control group, there was no difference in the rate of apoptosis of equal osmotic pressure group(P 〉 0.05),and the rate of apoptosis in HG group increased significantly( P 〈 0. 01 ). Compared with HG group, cell apoptosisin HG + LIRA group, HG + NAC group, HG + SB203580 group and HG+SP600125 group decreased significantly (P 〈 0. 01 ). Compared with the control group, the c-caspase-3 ,p-p38MAPK and p-JNK protein levels increased significantly in the HG group( P 〈 0.01 ). Compared with HG group, liraglutide significantly reduced the c-caspase-3, p-p38MAPK and p-JNK protein expression levels. Conclusion Liraglutide may inhibit high glucose-induced apoptosis in H9c2 cardiomyocyte through the inhibition of phosphorylation of p38MAPK and JNK.
出处
《临床内科杂志》
CAS
2017年第10期699-702,共4页
Journal of Clinical Internal Medicine
基金
武汉大学自主科研青年教师资助项目(2042017kf0157)
关键词
利拉鲁肽
心肌细胞凋亡
糖尿病心肌病
Liraglutide
Cardiomyocyte apoptosis
Diabetic cardiomyopathy