CAJ | 학술논문

目的：研究降钙素基因相关肽（CGRP）对血管平滑肌细胞（VSMCs）心肌素表达的影响及对细胞表型改变的调节作用。方法：取大鼠胸主动脉，以组织块贴壁培养法获得VSMCs，分为对照组、血管紧张素Ⅱ（ AngⅡ）组（加入AngⅡ处理）、CGRP 组（加入AngⅡ和CGRP 处理）和CGRP8－37组（在AngⅡ和 CGRP 基础上加入CGRP8－37）。 Western blot法检测VSMCs 中心肌素及细胞表型标志蛋白α－平滑肌肌动蛋白（α－SMA）和骨桥蛋白（ OPN）表达情况。结果：在VSMCs培养中，细胞心肌素表达水平随着培养时间延长逐渐降低，细胞培养48 h和72 h时心肌素表达水平较基线水平显著降低（ P＜0．05）；而加入CGRP处理后，心肌素表达水平逐渐增加，与基线水平比较，加入CGRP培养后48 h和72 h时细胞心肌素水平显著增加（ P＜0．05）。同时，在VSMCs培养48 h时，AngⅡ组细胞心肌素水平较对照组下降（P＜0．05），且α－SMA表达亦相应下降（P＜0．05），而OPN表达水平显著增加（P＜0．05）；CGRP组VSMCs心肌素水平较AngⅡ组增加，伴随着α－SMA表达水平增加（P＜0．05），相反地OPN表达水平下降（P＜0．05）；CGRP8－37组心肌素和α－SMA表达水平较CGRP＋AngⅡ组降低（P＜0．05），而OPN表达较CGRP组增加（ P＜0．05）。结论：CGRP通过促进VSMCs心肌素的表达而抑制细胞表型转换，使细胞维持收缩表型，且这一作用是CGRP通过与其受体结合后实现的。
목적：연구강개소기인상관태（CGRP）대혈관평활기세포（VSMCs）심기소표체적영향급대세포표형개변적조절작용。방법：취대서흉주동맥，이조직괴첩벽배양법획득VSMCs，분위대조조、혈관긴장소Ⅱ（ AngⅡ）조（가입AngⅡ처리）、CGRP 조（가입AngⅡ화CGRP 처리）화CGRP8－37조（재AngⅡ화 CGRP 기출상가입CGRP8－37）。 Western blot법검측VSMCs 중심기소급세포표형표지단백α－평활기기동단백（α－SMA）화골교단백（ OPN）표체정황。결과：재VSMCs배양중，세포심기소표체수평수착배양시간연장축점강저，세포배양48 h화72 h시심기소표체수평교기선수평현저강저（ P＜0．05）；이가입CGRP처리후，심기소표체수평축점증가，여기선수평비교，가입CGRP배양후48 h화72 h시세포심기소수평현저증가（ P＜0．05）。동시，재VSMCs배양48 h시，AngⅡ조세포심기소수평교대조조하강（P＜0．05），차α－SMA표체역상응하강（P＜0．05），이OPN표체수평현저증가（P＜0．05）；CGRP조VSMCs심기소수평교AngⅡ조증가，반수착α－SMA표체수평증가（P＜0．05），상반지OPN표체수평하강（P＜0．05）；CGRP8－37조심기소화α－SMA표체수평교CGRP＋AngⅡ조강저（P＜0．05），이OPN표체교CGRP조증가（ P＜0．05）。결론：CGRP통과촉진VSMCs심기소적표체이억제세포표형전환，사세포유지수축표형，차저일작용시CGRP통과여기수체결합후실현적。
AIM:Toinvestigatetheeffectsofcalcitoningene-relatedpeptide(CGRP)onmyocardinexpres-sion and phenotypic switch in vascular smooth muscle cells ( VSMCs) .METHODS:VSMCs were obtained by aortic tissue adherent culture and treated with angiotensin Ⅱ( AngⅡ) , AngⅡ+CGRP or AngⅡ+CGRP +CGRP8-37 .The protein expression of myocardin and the phenotypic proteins of the VSMCs was detected by Western blot .RESULTS:The expres-sion of myocardin in cultured VSMCs showed downregulation along with time expansion .The protein level of myocardin was higher at 48 h and 72 h than that at baseline in the cultured VSMCs (P<0.05).However, the myocardin was lower at 48 h and 72 h than that at baseline after treatment with CGRP in cultured VSMCs (P<0.05).Furthermore, at 48 h in cul-tured VSMCs, the myocardin decreased along with α-smooth muscle actin (α-SMA) (P<0.05), and osteopontin (OPN) increased (P<0.05) in AngⅡ group compared with control group .After treatment with CGRP, the levels of myocardin andα-SMA become higher ( P <0.05 ) but OPN was lower ( P <0.05 ) in CGRP group than those in AngⅡ group. CGRP8-37 abrogated CGRP-induced increase in myocardin and α-SMA and decrease in OPN in CGRP 8-37 group compared with CGRP group .CONCLUSION: CGRP may regulate the phenotypic switch of the VSMCs and maintain the cells in contractile phenotype through the upregulation of myocardin protein , which may be accomplished by the combination of CGRP and its receptor .