干扰素诱导蛋白p204过表达抑制大鼠主动脉血管外膜成纤维细胞凋亡、增殖与迁移

Interferon-induced Protein 204 Over-expression Inhibits Aortic Vascular Adventitial Fibroblast Proliferation and Migration in Experimental Rats

目的:观察干扰素诱导蛋白p204过表达对大鼠主动脉血管外膜成纤维细胞(VAFs)凋亡、增殖与迁移的影响。方法:分别应用携带p204基因(Ifi204)的慢病毒颗粒(Ifi204-Lv组)、空载慢病毒颗粒(Con-Lv组)感染VAFs,未经处理的VAFs作为阴性对照组。用噻唑蓝(MTT)比包法检测VAFs增殖,流式细胞术检测细胞凋亡,细胞划痕法和Transwell法测定细胞迁移情况,应用实时定量逆转录-聚合酶链反应(realtime q RT-PCR)和蛋白免疫印迹(Western blotting)分别检测p204、抑癌因子p53及细胞周期蛋白依赖性激酶抑制因子p21WAF(p21)的mRNA和蛋白表达。结果:Ifi204-Lv组与Con-Lv组、阴性对照组相比较,细胞增殖降低[(吸光度值),48 h:0.53±0.05 vs 0.66±0.03、0.63±0.06;72 h.0.89±0.06 vs 1.02±0.06、1.01±0.07]及迁移距离缩短[(像素),61.00±1.83 vs 74.50±6.25、75.50±7.85]、数量降低(61.75±10.69 vs 155.25±10.21、153.75±9.40),差异均有统计学意义(P均<0.05),但组问比较细胞凋亡率差异无统计学意义。与Con-Lv组、阴性对照组相比,Ifi204-Lv组的p204(3.45±0.15 vs 2.09±0.10、2.06±0.09)、p53(3.41±0.09 vs2.06±0.07、2.10±0.06)及p21(3.0l±0.08 vs2.05±0.06、2.11±0.08)的mRNA和蛋白表达均上调,差异均有统计学葸义(P均<0.05)。结论:p204过表达可抑制大鼠VAFs增殖与迁移,其机制可能部分与激活p53及p21表达有关。

Objective： To observe the effects of interferon-induced protein 204 （p204） over-expression on apoptosis, proliferation and migration of aortic vascular adventitial ifbroblast （VAFs） in experimental rats. Methods： Our research included in 3 groups： Iif204-Lv group, in whichVAFs were infected by Iif204-recombined lentivirus, Con-Lv group, in which VAFs carried the empty vector without virus, Blank control group, in which VAFs were untreated. VAFs proliferation was examined by MTT method, cell apoptosis was measured by lfow cytometry and the migration was detected by scratching assay and transwell chamber method. The mRNA and protein expressions of p204, p53 and p21were evaluated by real-time q RT-PCR and Western blot analysis respectively. Results： Compared with Con-Lv and Blank control groups, Iif204-Lv group had decreased VAFs proliferation （by OD value） at 48 hours： （0.53±0.05） vs （0.66±0.03） and （0.63 ± 0.06）, at 72 hours： （0.89 ± 0.06） vs （1.02 ± 0.06） and （1.01 ± 0.07）; distance of cell migration （by pixel）： （61.00 ± 1.83） vs （74.50 ± 6.25） and （75.50 ± 7.85）; number of cell migration： （61.75 ± 10.69） vs （155.25 ± 10.21） and （153.75 ± 9.40）, allP＆lt;0.05. VAFs apoptosis rates were similar among different groups. Compared with Con-Lv and Blank control groups, Ifi204-Lv group presented up-regulated mRNA＆amp;nbsp;expressions of p204 （3.45 ± 0.15） vs （2.09 ± 0.10） and （2.06 ± 0.09）; p53 （3.41 ± 0.09） vs （2.06 ± 0.07） and （2.10 ± 0.06）; p21 （3.01 ± 0.08） vs （2.05 ± 0.06） and （2.11 ± 0.08）, allP＆lt;0.05. Conclusion： p204 over-expression inhibits VAFs proliferation and migration which might be partly related to the activation of p53 and p21 expression in experimental rats.