摘要
目的观察微RNA-21(microRNA-21,miR-21)对p38丝裂原活化的蛋白激酶(p38mitogen-activated protein kinase,p38-MAPK)信号通路的调控作用,探究丹皮酚(paeonol,Pae)抑制血管内皮细胞(vascular endothelial cell,VEC)损伤的机制。方法组织块预消化贴壁法培养大鼠VEC;氧化低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导VEC的损伤;用HiPerFect试剂将miR-21模拟物或抑制剂转染进入VEC;免疫印迹法检测p38MAPK信号通路相关蛋白的表达;酶联免疫吸附试验检测VEC中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的水平。结果 ox-LDL明显提高VEC中Ras、p-MKK3/6、p-p38蛋白表达水平,并诱导VEC分泌TNF-α;miR-21高表达可升高TNF-α水平及Ras、p-MKK3/6、p-p38蛋白表达水平;Pae明显降低损伤的VEC分泌TNF-α水平,且抑制由miR-21高表达引起的TNF-α水平上升及p38MAPK信号通路的激活。结论 Pae可抑制miR-21介导的p38MAPK信号通路,减少ox-LDL诱导的VEC中TNF-α的分泌。
Objective To observe the regulatory effect of microRNA-21 （rniR-21） on the p38 mitogen-activated protein kinase （p38 MAPK） pathway and to investigate the molecular mechanism by which paeonol （Pae） inhibits injury of vascular endothelial cells （VECs）. Methods VECs were isolated from rat thoracic aortas by collagenase digestion and adherent culture and then injured by oxidized low-density lipoprotein （ox-LDL） ;the cells were transfected with miR-2] mimic or inhibitor using a HiPerFect transfection reagent. The protein expression of Ras, p-MKK3/6, and p-p38 was measured by Western blotting; the level of tumor necrosis factor-α （TNF-α） in VECs was measured by enzyme-linked immunosorbent assay. Results The ox-LDL significantly increased the protein expression of Ras, p-MKK3/6, and p- p38 and induced secretion of TNF-α in VECs. Enhanced miR-21 expression increased the level of TNF-α and protein expression of Ras,p-MKK3/6 ,and p-p38. Pae significantly reduced the release of TNF-α in injured VECs and inhibited the increase in TNF-α level and activation of p38 MAPK pathway caused by enhanced miR-21 expression. Conclusion Pae can inhibit the rrflR-21-mediated p38 MAPK pathway and reduce ox-LDL-induced release of TNF-α in VtK;s.
出处
《安徽中医药大学学报》
2014年第1期51-56,共6页
Journal of Anhui University of Chinese Medicine
基金
国家自然科学基金项目(81073090
81274134)
安徽中医药大学研究生科技创新基金(2013zd003)
