利拉鲁肽通过p38丝裂原活化蛋白激酶和c-Jun氨基末端激酶通路抑制高糖培养的H9e2细胞凋亡

Liraglutide inhibits high glucose-induced apoptosis in H9c2 cardiomyocyte through the p38 mitogen activated protein ldnase and c-Jun N-termlnal kinase pathway

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.