OX40-OX40L相互作用对小鼠平滑肌细胞亲环素A表达的影响  

Effect of OX40-OX40 Ligand Interaction on the Expression of Cyclophilin A in Mice Vascular Smooth Muscle Cells

作　　者：逯朝阳 严金川[1] 伍超[1] 徐丽华[1] 王中群[1] 袁伟[1] 

机构地区：[1]江苏大学附属医院心内科,江苏省镇江市212001

出　　处：《中国动脉硬化杂志》2014年第6期558-562,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金(81170279;81370409);江苏省自然科学基金(BK2011486);江苏省科教兴卫工程(LJ201116);镇江市心血管病重点实验室项目(SS2012002)

摘　　要：目的观察OX40-OX40配体(OX40L)相互作用对小鼠主动脉血管平滑肌细胞(VSMC)亲环素A(CyPA)表达的影响。方法小鼠VSMC采用组织块贴壁法原代培养;应用实时荧光定量PCR和流式细胞术检测OX40L mRNA和蛋白表达水平;细胞内、外CyPA水平分别采用实时荧光定量PCR及Western blot方法检测。结果肿瘤坏死因子α+脂多糖联合处理小鼠VSMC,细胞OX40L mRNA和蛋白表达呈时间依赖性,mRNA表达在24 h(5.967±0.252比1.000±0.000,P<0.05)达到高峰,蛋白表达在36 h(61.900±2.551比20.967±0.451,P<0.05)增加最显著;可溶性OX40处理VSMC后,CyPA mRNA表达呈时间依赖性并在90 min达到高峰(1.799±0.098比1.000±0.000,P<0.05),2 h后细胞培养上清中可检测到高水平的CyPA(1.119±0.059比0.281±0.038,P<0.05),而细胞内CyPA表达较对照组无明显差异;抗OX40L预处理细胞可显著抑制可溶性OX40诱导的CyPA mRNA(1.105±0.091比1.799±0.098,P<0.05)及蛋白(0.635±0.040比1.119±0.059,P<0.05)表达。结论 OX40-OX40L相互作用可影响VSMC CyPA的分泌。Aim To observe the effect of OX40-OX40 ligand （OX40L） interaction on the expression of cyclo- philin A （CyPA） in mice aortic vascular smooth muscle cell （VSMC）. Methods Patch-attaching method was used for primary culture of mouse aortic VSMC. The expression of OX40L mRNA and protein in VSMC were measured by real- time quantitative PCR （RT-PCR） and flow cytometry （ FCM）, respectively. The levels of CyPA inside and outside of the VSMC was determined by RT-PCR and Western blot. Results The expression of OX40L mRNA and protein was time- dependently induced by tumor necrosis factor-or （TNF-ot） plus lipopolysaccharide （LPS）, the mRNA expression peaked at 24 h （5. 967 ± 0. 252 vs 1. 000± 0. 000, P 〈 0. 05 ）, and the expression of protein peaked at 36 h （61. 900 ± 2. 551 vs 20. 967±0. 451, P 〈 0. 05 ）. The expression of CyPA mRNA was time-dependently induced by soluble OX40 （ sOX40）, and peaked at 90 minute （ 1. 799±0. 098 vs 1. 000±0. 000, P 〈 0. 05）. And abundant CyPA was gained in supernatant at 2 h （ 1.119 ± 0. 059 vs 0. 281 s 0. 038, P 〈 0. 05）, whereas, we didn＇ t observe the change of CyPA protein inside the VSMC compared to the control. Notably, the sOX40 revulsive elevations of mRNA （ 1. 105 ± 0. 091 vs 1. 799 ± 0. 098, P 〈0. 05） and protein （0. 635 ±0. 040 vs 1. 119 ±0. 059, P 〈0. 05） can be blocked by anti-OX40L monoclonal antibody. Conclusion OX40-OX40 ligand interaction could affect the secretion of CyPA in VSMC.

关 键 词：OX40配体 血管平滑肌细胞 亲环素A 动脉粥样硬化 

分 类 号：R392[医药卫生—免疫学]