p38 MAPK信号通路在尼古丁诱导的血管内皮细胞自噬中的作用(1)
【摘要】 目的：探讨丝裂原活化蛋白激酶(MAPK)通路在尼古丁诱导人脐静脉内皮细胞(HUVECs)自噬中的作用。方法：不同浓度尼古丁(6×10-9、6×10-8、6×10-7、6×10-6 mol/L)处理HUVECs，采用CCK-8和LDH试剂盒检测尼古丁对血管内皮细胞增殖和活性的影响，运用蛋白免疫印迹(Western blot)技术检测尼古丁作用下Beclin-1和LC3Ⅱ的表达及ERK1/2、JNK、p38 MAPK的磷酸化水平。
结果：与正常对照组相比，较高浓度尼古丁(≥6×10-7 mol/L)抑制HUVECs的增殖能力(P<0.01)，增加上清中LDH的活性(P<0.05)，且在尼古丁浓度为6×10-6 mol/L时作用最明显。与正常对照组相比，尼古丁(6×10-6 mol/L)作用下LC3Ⅱ表達显著增加(P<0.01)，而Beclin-1的表达减少(P<0.05)；同时尼古丁磷酸化MAPK信号，其中ERK1/2和p38 MAPK的磷酸化水平分别增加了0.77倍和2.91倍(P<0.01)，而JNK的磷酸化水平没有明显变化。与尼古丁组相比，SB+尼古丁组中LC3Ⅱ与Beclin-1蛋白表达均增加(P<0.05)。结论：尼古丁可导致人脐静脉内皮细胞损伤并抑制细胞自噬，此过程中p38 MAPK信号通路可能起了重要作用。
, 百拇医药
【关键词】 人脐静脉内皮细胞； 尼古丁； 自噬； 心血管疾病； MAPK
The Role of p38 MAPK Signaling Pathway in Nicotine-induced Vascular Endothelial Cells Autophagy/WANG Ruiling，LIU Wenxia，LIU Caihong，et al.//Medical Innovation of China，2018，15(26)：0-045
【Abstract】 Objective：To investigate the differential role of mitogen-activated protein kinase(MAPK) pathway signaling in nicotine-induced vascular endothelial cells autophagy.Method：HUVECs were treated with different doses of nicotine(6×10-9，6×10-8，6×10-7，6×10-6 mol/L) .The effect of nicotine on the proliferation and activity of vascular endothelial cells was detected by CCK-8 and LDH kit.The protein expression level of Beclin-1 and LC3Ⅱand the phosphorylation of ERK1/2、p38 and JNK induced by nicotine were detected by Western blot.Result：Compared to the control group，the results indicated that higher doses of nicotine(≥6×10-7 mol/L) inhibited the ability of cell proliferation(P<0.01)，and increased the activity of supernatant LDH (P<0.05)，especially the 6×10-6 mol/L of nicotine had obvious effects.Compared with normal control group，nicotine(6×10-6 mol/L) significantly increased the LC3Ⅱ protein expression(P<0.01)，while decreased the protein expression of Beclin-1(P<0.05).Meanwhile，nicotine contributed to the phosphorylation of MAPK，among them，the phosphorylation of ERK1/2 and p38 MAPK increased by 0.77-fold and 2.91-fold(P<0.01)，while the phosphorylation of JNK had no change.Compared to nicotine group， the LC3Ⅱand Beclin-1 protein expression of SB+nicotine group all increased(P<0.05).Conclusion：Nicotine can damage human umbilical vein endothelial cells and inhibit autophagy，and p38 MAPK signaling pathway may play an important role in this process.
【Key words】 HUVECs； Nicotine； Autophagy； Cardiovascular disease； MAPK
First-author’s address：Shanxi Medical University，Taiyuan 030001，China
doi：10.3969/j.issn.1674-4985.2018.26.010, 百拇医药(王瑞玲 刘雯霞 刘彩红 郭蕊 王亚静)
结果：与正常对照组相比，较高浓度尼古丁(≥6×10-7 mol/L)抑制HUVECs的增殖能力(P<0.01)，增加上清中LDH的活性(P<0.05)，且在尼古丁浓度为6×10-6 mol/L时作用最明显。与正常对照组相比，尼古丁(6×10-6 mol/L)作用下LC3Ⅱ表達显著增加(P<0.01)，而Beclin-1的表达减少(P<0.05)；同时尼古丁磷酸化MAPK信号，其中ERK1/2和p38 MAPK的磷酸化水平分别增加了0.77倍和2.91倍(P<0.01)，而JNK的磷酸化水平没有明显变化。与尼古丁组相比，SB+尼古丁组中LC3Ⅱ与Beclin-1蛋白表达均增加(P<0.05)。结论：尼古丁可导致人脐静脉内皮细胞损伤并抑制细胞自噬，此过程中p38 MAPK信号通路可能起了重要作用。
, 百拇医药
【关键词】 人脐静脉内皮细胞； 尼古丁； 自噬； 心血管疾病； MAPK
The Role of p38 MAPK Signaling Pathway in Nicotine-induced Vascular Endothelial Cells Autophagy/WANG Ruiling，LIU Wenxia，LIU Caihong，et al.//Medical Innovation of China，2018，15(26)：0-045
【Abstract】 Objective：To investigate the differential role of mitogen-activated protein kinase(MAPK) pathway signaling in nicotine-induced vascular endothelial cells autophagy.Method：HUVECs were treated with different doses of nicotine(6×10-9，6×10-8，6×10-7，6×10-6 mol/L) .The effect of nicotine on the proliferation and activity of vascular endothelial cells was detected by CCK-8 and LDH kit.The protein expression level of Beclin-1 and LC3Ⅱand the phosphorylation of ERK1/2、p38 and JNK induced by nicotine were detected by Western blot.Result：Compared to the control group，the results indicated that higher doses of nicotine(≥6×10-7 mol/L) inhibited the ability of cell proliferation(P<0.01)，and increased the activity of supernatant LDH (P<0.05)，especially the 6×10-6 mol/L of nicotine had obvious effects.Compared with normal control group，nicotine(6×10-6 mol/L) significantly increased the LC3Ⅱ protein expression(P<0.01)，while decreased the protein expression of Beclin-1(P<0.05).Meanwhile，nicotine contributed to the phosphorylation of MAPK，among them，the phosphorylation of ERK1/2 and p38 MAPK increased by 0.77-fold and 2.91-fold(P<0.01)，while the phosphorylation of JNK had no change.Compared to nicotine group， the LC3Ⅱand Beclin-1 protein expression of SB+nicotine group all increased(P<0.05).Conclusion：Nicotine can damage human umbilical vein endothelial cells and inhibit autophagy，and p38 MAPK signaling pathway may play an important role in this process.
【Key words】 HUVECs； Nicotine； Autophagy； Cardiovascular disease； MAPK
First-author’s address：Shanxi Medical University，Taiyuan 030001，China
doi：10.3969/j.issn.1674-4985.2018.26.010, 百拇医药(王瑞玲 刘雯霞 刘彩红 郭蕊 王亚静)