High glucose induces apoptosis in AC16 human cardiomyocytes via macrophage migration inhibitory factor and c-Jun N-terminal kinase
High glucose induces apoptosis in AC16 human cardiomyocytes via macrophage migration inhibitory factor and c-Jun N-terminal kinase
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高糖通过巨噬细胞迁移抑制因子和c-Jun N端激酶诱导AC16人心肌细胞凋亡
DOI：
10.1111/j.1440-1681.2010.05420.x
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发表时间：
2010-10-01
影响因子：
2.9
通讯作者：
Yu, Xi-Yong
中科院分区：
文献类型：
--
作者：
Liang, Jia-Liang;Xiao, Ding-Zhang;Yu, Xi-Yong
已知高糖可诱导心肌细胞凋亡，巨噬细胞移动抑制因子(MIF)可能参与糖尿病的发生发展。然而，糖尿病心肌病中高糖与MIF之间的关系尚不清楚。在本研究中，AC16人心肌细胞在25 mmol/L葡萄糖中培养20、30和60min，然后进行蛋白质印迹分析以检测MIF的表达和c-jun氨基末端激酶(JNK)的激活。此外，用2.5mU/L SP600125(JNK抑制剂)、40mU/L(S，r)-3-(4-hydroxyphenyl)-4，5-dihydro-5-isoxazole乙酸甲酯，MIF拮抗剂)或0.1%二甲基亚砜(DMSO；载体)处理AC16细胞1h后，加入25 mmoL/L葡萄糖，培养72h，然后进行Annexin V-异硫氰酸荧光素/碘化丙啶染色和流式细胞仪分析。免疫印迹法检测caspase3活性和JNK的磷酸化水平。高浓度葡萄糖使AC16心肌细胞内源性MIF和JNK磷酸化表达增加。SP600125和ISO-1可降低葡萄糖诱导的细胞凋亡和caspase3活性。此外，抑制内源性MIF可抑制JNK的磷酸化。综上所述，高糖诱导的心肌细胞凋亡涉及MIF的过度表达和JNK信号通路的激活。高糖-MIF-JNK通路的发现将有助于确定糖尿病心肌病治疗的潜在新靶点。
P>1. It is known that high glucose can induce cardiomyocyte apoptosis and that macrophage migration inhibitory factor (MIF) may be involved in the development of diabetes. However, the relationship between high glucose and MIF in diabetic cardiomyopathy remains unclear.2. In the present study, AC16 human cardiomyocytes were cultured in the presence of 25 mmol/L glucose for 20, 30 and 60 min before being subjected to western blot analyses to determine MIF expression and c-Jun N-terminal kinase (JNK) activation. In addition, AC16 cells were pretreated with 2.5 mu mol/L SP600125 (a JNK inhibitor), 40 mu mol/L (s,r)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester (ISO-1; an MIF antagonist) or 0.1% dimethylsulphoxide (DMSO; vehicle) for 1 h prior to exposure to 25 mmol/L glucose and culture for 72 h, followed by annexin V-fluorescein isothiocyanate/propidium iodide staining and flow cytometry analysis. Caspase 3 activity and phosphorylation of JNK were also analysed by western blotting.3. The high concentration of glucose increased expression of endogenous MIF and JNK phosphorylation in AC16 cardiomyocytes. Pretreatment of cells with SP600125 and ISO-1 reduced glucose-induced apoptosis and caspase 3 activity. Furthermore, JNK phosphorylation was attenuated by inhibition of endogenous MIF.4. In conclusion, myocardial cell apoptosis induced by high glucose involves the overexpression of MIF and activation of the JNK signalling pathway. The identification of a high glucose-MIF-JNK pathway will help determine potential new targets in the treatment of diabetic cardiomyopathy.