Involvement of JNK1/2-NF-κBp65 in the regulation of HMGB2 in myocardial ischemia/reperfusion-induced apoptosis in human AC16 cardiomyocytes
Involvement of JNK1/2-NF-κBp65 in the regulation of HMGB2 in myocardial ischemia/reperfusion-induced apoptosis in human AC16 cardiomyocytes
复制标题
JNK1/2-NF-κBp65参与HMGB2在T心肌缺血/再灌注诱导的人AC16心肌细胞凋亡中的调节
DOI：
10.1016/j.biopha.2018.07.015
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发表时间：
2018-10-01
影响因子：
7.5
通讯作者：
Xu, Yawei
中科院分区：
文献类型：
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作者：
Zhang, Wei;Zhang, Yangyang;Xu, Yawei
JNK1/2和核因子-kappaB信号是介导多种细胞过程的重要信号通路，包括细胞存活、细胞凋亡、炎症和血管生成。在缺血再灌注(I/R)诱导的损伤中，发现JNK1/2活化和核转录因子-kappaBp65核转位。然而，JNK1/2-NF-kappa Bp65信号通路的调控机制尚不清楚。为探讨HMGB2在细胞I/R损伤中的作用及可能机制，用SP600125(JNK1/2抑制剂)或PDTC(核因子-kappa B抑制剂)处理转染pLVX-Puro-HMGB2的AC16心肌细胞，并在I/R损伤后再用HMGB2-shRNA转染。CCK-8法检测细胞增殖，流式细胞仪检测细胞凋亡率，TUNEL法检测细胞凋亡率。Western印迹法检测HMGB2、裂解PARP和Caspase-3、Bax和Bcl2的表达及MAPKs和NF-kappa Bp65的活性。在此，我们发现I/R可时间依赖性地诱导AC16心肌细胞HMGB2的表达增加。HMGB2沉默可显著抑制I/R诱导的细胞增殖抑制、细胞凋亡、ERK1/2、JNK1/2和NF-kappaBp65的活化，增加Bax，裂解PARP和Caspase-3，降低Bcl2的表达。HMBG2过表达与AC16心肌细胞I/R损伤相似，SP600125或PDTC可逆转HMBG2过表达。此外，PDTC对大鼠脑I/R损伤后的细胞凋亡、HMGB2、裂解的PARP、Caspase-3和Bax的表达以及JNK1/2的激活也有抑制作用。综上所述，我们的研究结果表明，HMBG2的过表达通过激活AC16心肌细胞中的JNK1/2-NF-kappa Bp65信号来促进I/R诱导的细胞凋亡。
JNK1/2 and NF-kappa B signal are essential signaling pathways that mediate a variety of cellular processes, including cell survival, apoptosis, inflammation and angiogenesis. JNK1/2 activation and NF-kappa Bp65 nuclear translocation have been found in ischemia/reperfusion (I/R)-induced injury. However, the regulation of JNK1/2-NF-kappa Bp65 signaling pathway remains unclear. To examine the function and possible mechanism of HMGB2 in I/R-induced cell injury, human AC16 cardiomyocytes transfected with pLVX-Puro-HMGB2 were treated with SP600125 (JNK1/2 inhibitor) or PDTC (NF-kappa B inhibitor) and that following I/R injury were transfected with HMGB2-shRNA. The cell proliferation and apoptosis were measured by CCK-8, flow cytometry and TUNEL, respectively. The expression of HMGB2, Cleaved PARP and Caspase-3, Bax and Bcl-2 and activity of MAPKs and NF-kappa Bp65 were measured by Western blot. Here, we found that I/R time-dependently induced the increase in the expression of HMGB2 in AC16 cardiomyocytes. HMGB2 silencing significantly inhibited I/R-induced the cell proliferation reduction, cell apoptosis, activation of ERK1/2, JNK1/2 and NF-kappa Bp65, increased Bax, Cleaved PARP and Caspase-3 and decreased Bcl-2 expression. HMBG2 overexpression mimicked the effect of I/R-induced injury in AC16 cardiomyocytes, which was reversed by treatment with SP600125 or PDTC. Moreover, PDTC treatment in rats following I/R injury also showed decreases in the cell apoptosis, HMGB2, Cleaved PARP and Caspase-3 and Bax expression, and JNK1/2 activation. Taken together, our findings suggest that HMBG2 overexpression promotes I/R-induced cell apoptosis through activating the JNK1/2-NF-kappa Bp65 signaling in AC16 cardiomyocytes.