摘要
目的观察丹皮酚(Paeonol,Pae)对脂多糖(lipopolysaccharide,LPS)损伤的大鼠血管内皮细胞(vascular endothelial cells,VECs)炎性因子释放及对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)凋亡的影响,阐明Pae抑制VSMCs凋亡是否通过影响VECs释放的炎性因子调控p38 MAPK信号通路。方法组织块预消化贴壁法原代培养VECs和VSMCs;Transwell小室建立VSMCs和VECs共培养体系;LPS诱导VECs损伤;MTT方法检测细胞存活率;ELISA检测VECs分泌肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)的水平;流式细胞仪检测VSMCs凋亡率;Western blotting检测VSMCs中p38MAPK信号通路及凋亡相关蛋白表达。结果与正常对照组比较,LPS可显著提高VECs分泌的TNF-α水平(P<0.05),明显升高共培养体系中VSMCs p38 MAPK信号通路蛋白及凋亡相关蛋白(p-MKK3/6、pp38、p53和Cleaved caspase-3)水平(P<0.05),显著提高VSMCs凋亡率(P<0.05);与LPS刺激组比较,Pae可显著抑制VECs分泌的TNF-α(P<0.05),明显降低共培养体系中VSMCs p38MAPK信号通路蛋白及凋亡相关蛋白(p-MKK3/6、p-p38、p53和Cleaved caspase-3)水平(P<0.05),显著降低VSMCs凋亡率(P<0.05)。结论 Pae可抑制VECs释放TNF-α,从而抑制p38MAPK信号通路,进而减少VSMCs凋亡。
Objective To investigate the effects of paeonol （Pae） on the release of inflammatory factors from rat vascular endothelial cells （VECs） with lipopolysaccharide （LPS）-induced injury and the apoptosis of vascular smooth muscle cells （VSMCs）, as well as whether Pae inhibits the apoptosis of VSMCs by affecting the release of inflammatory factors from VECs and regulating the p38 MAPK signaling pathway. Methods The tissue predigested adherent method was used for the primary culture of VECs and VSMCs. The Transwell chamber was used to establish a co-culture system of VSMCs and VECs. LPS was used to induce the injury of VECs. The MTT assay was used to measure the survival rate of cells; ELISA was used to measure the level of tumor necrosis factor-α （TNF-α） secreted by VECs; flow cytometry was used to measure the apoptosis rate of VSMCs; Western blot was used to measure the expression of proteins involved in the p38 MAPK signaling pathway and apoptosis-related proteins in VSMCs. Results Compared with the normal control group, LPS significantly increased the level of TNF-α in VECs （P〈0.05）, the expression of the p38 MAPK signaling pathway proteins and apoptosis-related proteins （p-MKK3/6, p-p38, p53, and cleaved caspase-3） in VSMCs in the co-culture system （P〈0.05）, and the apoptosis rate of VSMCs （P〈0.05）. Compared with the LPS stimulation group, Pae significantly inhibited the secretion of TNF-α from VECs （P〈0.05） and significantly reduced the expression of the p38 MAPK signaling pathway proteins and apoptosis-related proteins （p-MKK3/6, p-p38, p53, and cleaved caspase-3） in VSMCs in the co-culture system （P〈0.05） and the apoptosis rate of VSMCs （P〈0.05）. Conclusion Pae can inhibit the release of TNF-α from VECs and the p38 MAPK signaling pathway and thus reduce the apoptosis of VSMCs.
作者
刘雅蓉
吴鸿飞
戴敏
LIU Ya-rong;WU Hong-fei;DAI Min(School of Pharmacy,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;Key Laboratory of Xin an Medicine,Ministry of Education,Anhui Hefei 230038,China;Anhui Province Key Laboratory of R＆D of Chinese Medicine,Anhui Hefei 230012,China)
出处
《安徽中医药大学学报》
2018年第4期65-71,共7页
Journal of Anhui University of Chinese Medicine
基金
国家自然科学基金项目(81773937
81473386)
安徽中医药大学自然科学研究项目(2018zrzd06)
作者简介
刘雅蓉（1989-），女，硕士，助教;通信作者： 戴敏，daiminliao@163.com
