CAJ | 학술논문

目的探讨肝细胞生长因子(HGF)和转化生长因子β1(TGFβ1)对人心房成纤维细胞α-平滑肌肌动蛋白(α-SMA)和胶原蛋白Ⅰ表达的影响,探究其调控心房颤动(AF)患者心房纤维化的分子机制.方法 20例风湿性心脏病(rheumatic heart disease,RHD)接受换瓣手术患者,收集其临床资料.窦性心律(SR)组10例；慢性心房颤动(CAF)组10例.取有心耳组织,体外分离培养人心房成纤维细胞,分别给予HGF和TGFβ1干预,以半定量逆转录-聚合酶链反应(RT-PCR)技术检测细胞Ⅰ型胶原和0-SMA的mRNA含量,以免疫荧光、免疫印迹技术(Western blot)检测细胞α-SMA蛋白水平.结果 (1)与SR组比较,CAF组左心房较大(t=2.692,P＜0.05),成纤维细胞Ⅰ型胶原和α-SMA的mRNA水平较高(P ＜0.01);CAF组Ⅰ型胶原的mRNA水平与左心房内径(LAD)(r=0.836,P =0.014)、AF持续时间(r=0.739,P=0.045)、α-SMA的mRNA水平(r=0.886,P=0.012)呈明显正相关.(2)与SR组比较,CAF组细胞α-SMA蛋白表达水平较高(P＜0.01).(3)CAF组心房成纤维细胞予TGFβ1干预后,Ⅰ型胶原、α-SMA mRNA及α-SMA蛋白表达明显增加(P＜0.01)；予HGF干预后,Ⅰ型胶原、α-SMA mRNA及α-SMA蛋白表达明显减少(P＜0.01).结论 AF患者心房成纤维细胞中Ⅰ型胶原和α-SMA分泌增加,导致心房结构重构,可能与AF的发生和维持有关.TGFβ1上调人成纤维细胞中α-SMA和Ⅰ型胶原表达,促进心房纤维化；而HGF阻止人心房纤维化进程.
목적 탐토간세포생장인자(HGF)화전화생장인자β1(TGFβ1)대인심방성섬유세포α-평활기기동단백(α-SMA)화효원단백Ⅰ표체적영향,탐구기조공심방전동(AF)환자심방섬유화적분자궤제.방법 20례풍습성심장병(rheumatic heart disease,RHD)접수환판수술환자,수집기림상자료.두성심률(SR)조10례；만성심방전동(CAF)조10례.취유심이조직,체외분리배양인심방성섬유세포,분별급여HGF화TGFβ1간예,이반정량역전록-취합매련반응(RT-PCR)기술검측세포Ⅰ형효원화0-SMA적mRNA함량,이면역형광、면역인적기술(Western blot)검측세포α-SMA단백수평.결과 (1)여SR조비교,CAF조좌심방교대(t=2.692,P＜0.05),성섬유세포Ⅰ형효원화α-SMA적mRNA수평교고(P ＜0.01);CAF조Ⅰ형효원적mRNA수평여좌심방내경(LAD)(r=0.836,P =0.014)、AF지속시간(r=0.739,P=0.045)、α-SMA적mRNA수평(r=0.886,P=0.012)정명현정상관.(2)여SR조비교,CAF조세포α-SMA단백표체수평교고(P＜0.01).(3)CAF조심방성섬유세포여TGFβ1간예후,Ⅰ형효원、α-SMA mRNA급α-SMA단백표체명현증가(P＜0.01)；여HGF간예후,Ⅰ형효원、α-SMA mRNA급α-SMA단백표체명현감소(P＜0.01).결론 AF환자심방성섬유세포중Ⅰ형효원화α-SMA분비증가,도치심방결구중구,가능여AF적발생화유지유관.TGFβ1상조인성섬유세포중α-SMA화Ⅰ형효원표체,촉진심방섬유화；이HGF조지인심방섬유화진정.
Objective To investigate the effect of hepatocyte growth factor (HGF) and transforming growth factor-β1 (TGFβ1) on the expression of α-smooth muscle actin (α-SMA) and collagen Ⅰ in human atrial fibroblast in vitro,and to explore the possible molecular mechanism of atrial fibrosis in patients with atrial fibrillation (AF).Methods Human atrial fibroblast,isolated from aseptic right atrial appendage tissues of 10 sinus rhythm (SR) and 10 chronic atrial fibrillation (CAF) patients,were cultured with HGF and TGFβ1.mRNA expressions of collagen Ⅰ and α-SMA were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR),the protein expression of α-SMA was determined by immunofluorescence and Western blot.Results (1) Compared with SR group,left atrium was significantly dilated in CAF group(t =2.692,P ＜0.05),the mRNA expression of collagen Ⅰ and α-SMA of atrial fibroblasts were significantly upregulated (all P ＜ 0.01),mRNA expression of collagen Ⅰ was positively correlated with left atrial dimension (LAD) (r =0.836,P =0.014),AF duration (r =0.739,P =0.045) and α-SMA mRNA level (r =0.886,P =0.012).(2) Compared with SR group,the expression of α-SMA protein in CAF atrial fibroblasts were significantly increased (P ＜ 0.01).(3) TGFβ1 further stimulated while HGF significantly attenuated the expression of collagen Ⅰ and α-SMA in CAF atrial fibroblasts (all P ＜0.01).Conclusions Increasing expression of collagen Ⅰ and α-SMA in human atrial fibroblasts might promote atria remodeling leading to the development and sustaining of AF.HGF is involved in the negative regulation on the expression of α-SMA and collagen Ⅰ.