摘要
目的探讨降钙素基因相关肽(calcitonin gene-related peptide,CGRP)对体外培养的血管平滑肌细胞(vascular smooth muscle cells,VSMCs)心肌素表达及细胞炎症激活的影响。方法取大鼠胸主动脉,以组织块贴壁培养法获得VSMCs,根据不同的处理方案,分为对照组、IL-1β组、CGRP组、心肌素高表达组、心肌素干扰组和CGRP8-37组(CGRP受体拮抗剂组)。Western blot检测不同组VSMCs中心肌素和IL-6蛋白表达水平。结果以IL-1β处理VSMCs后,细胞培养24、48、72 h时IL-6表达水平均高于基线水平(P<0.05),而心肌素水平无显著变化;以高表达心肌素的腺病毒载体转染VSMCs并给予IL-1β处理后,与单纯IL-1β处理比较,各时间点IL-6水平显著下降(P<0.05)。CGRP组中各个时间点VSMCs内心肌素水平较同时间点IL-1β组心肌素水平显著增加[24 h:(1.17±0.03)vs(0.25±0.03);48 h:(2.04±0.05)vs(0.19±0.01);72 h:(0.96±0.02)vs(0.28±0.02);P<0.05],而细胞内IL-6表达水平较同时间点IL-1β组水平显著降低[24 h:(0.23±0.01)vs(0.77±0.03);48 h:(0.28±0.02)vs(1.35±0.04);72 h:(0.20±0.02)vs(1.04±0.04);P<0.05]。心肌素干扰组VSMCs中CGRP处理后各个时间点细胞内IL-6水平较CGRP组显著升高[24 h:(1.74±0.04)vs(0.51±0.01);48 h:(1.68±0.03)vs(0.29±0.01);72 h:(0.98±0.02)vs(0.31±0.01);P<0.05],且CGRP8-37组与CGRP组比较也获得类似结果,而心肌素干扰组和CGRP8-37组中同时间点的细胞内IL-6水平无明显差异。结论 CGRP通过促进VSMCs心肌素的表达而抑制IL-1β诱导的细胞炎症激活蛋白IL-6表达,且这一作用是CGRP通过与其受体结合后实现的。
Objective To investigate the effect of calcitonin gene-related peptide( CGRP) on myocardin expression and inflammatory activation in vascular smooth muscle cells( VSMCs) in vitro.Methods VSMCs were obtained by aortic tissue adherent culture and treated with interleukin-1β( IL-1βgroup),IL-1β + CGRP( CGRP group),and IL-1β + CGRP + CGRP8-37( CGRP receptor antagonist,CGRP8-37group) or transfected with recombinant adenovirus vector over-expressing myocardin( myocardin group) or siRNA-myocardin( siRNA-myocardin group). The expression of myocardin protein and interleukin-6( IL-6)protein at different time points in the VSMCs were detected by Western blotting. Results In the in vitro cultured VSMCs of the IL-1β group,the expression of IL-6 at 24,48 and 72 h were significantly higher than the baseline( P〈0. 05),and the expression of myocardin was similar. In the myocardin group,the level of IL-6 at any time point was obviously lower than that in the IL-1β group( P〈0. 05). The CGRP group showed higher myocardin expression and lower IL-6 level at any time point than the IL-1β group( myocardin: 1. 17 ±0. 03 vs 0. 25 ± 0. 03 at 24 h; 2. 04 ± 0. 05 vs 0. 19 ± 0. 01 at 48 h; 0. 96 ± 0. 02 vs 0. 28 ± 0. 02 at 72 h; P〈0. 05; IL-6: 0. 23 ± 0. 01 vs 0. 77 ± 0. 03 at 24 h; 0. 28 ± 0. 02 vs 1. 35 ± 0. 04 at 48 h; 0. 20 ± 0. 02 vs1. 04 ± 0. 04 at 72 h; P〈0. 05). The level of IL-6 in the siRNA-myocardin group was higher than that in the IL-1β group at any time point( 1. 74 ± 0. 04 vs 0. 51 ± 0. 01 at 24 h; 1. 68 ± 0. 03 vs 0. 29 ± 0. 01 at 48 h;0. 98 ± 0. 02 vs 0. 31 ± 0. 01 at 72 h; P〈0. 05). The comparison had similar results between the CGRP8-37 group and the CGRP group. Also,there was no difference in the IL-6 level at the same time point between the siRNA-myocardin group and the CGRP8-37 group. Conclusion CGRP may inhibit IL-1β-induced IL-6 protein expression through up-regulating the expression of myocardin protein in the VSMCs, which may be accomplished by binding CGRP and its receptor.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2015年第20期2027-2031,共5页
Journal of Third Military Medical University
基金
国家自然科学基金(81360021)
贵州省国际合作项目[黔科合外G字(2013)7037号]~~
关键词
血管平滑肌细胞
降钙素基因相关肽
心肌素
白介素-6
vascular smooth muscle cells
calcitonin gene-related peptide
myocardin
interleukin-6