CAJ | 학술논문

目的：探讨阿司匹林（aspirin，aspi）对醛固酮（ald）诱导的新生大鼠心肌成纤维细胞（cardiac fibroblasts， CF）增殖的影响及可能机制。方法0．08％胰酶消化1～3 d SD新生大鼠，差速贴壁法纯化SD新生大鼠原代CF。正式实验采用培养3～5代CF，分为4组，空白对照组：无小牛血清的高糖DMEM；醛固酮组（ ald）：无小牛血清的高糖DMEM＋1×10－8 mol／L ald；阿司匹林＋醛固酮组（ aspi＋ald）：无小牛血清的高糖DMEM＋1×10－8 mol／L ald ＋1．11×10－6 mol／L aspi；螺内酯＋醛固酮组（ spiro ＋ald ）：无小牛血清的高糖 DMEM ＋1×10－8 mol／L ald＋1×10－6 mol／L spiro。 HE染色法观察各组CF形态学变化；四氮唑盐（ MTT）比色法检测细胞增殖；Western印迹分析TGF-β-Smad 2、3、4的蛋白表达。结果 HE染色结果显示，与对照组相比，ald可使细胞增生活跃，分裂相细胞显著增多（P＜0．01）；与ald相比，Aspi＋ald组及spiro＋ald组均可使细胞分裂显著减少（P＜0．01）。 MTT法检测细胞增殖结果提示，与对照组相比，ald能够明显增加CF对MTT的代谢率，MTT值明显升高，与对照组相比差异显著（P＜0．01）；与 ald 组相比，aspi ＋ald 组、spiro ＋ald 组均可使CF 对MTT 代谢率减少，MTT值显著降低（P＜0．01）。 Western印迹检测结果显示，以ald刺激CF后，TGF-β-Smad 2、Smad 3及Smad 4蛋白水平明显增高，以aspi及spiro干预后，CF中TGF-β-Smad 2、Smad 3及Smad 4蛋白水平表达较ald组明显降低，均有显著性差异（ P＜0．05）。结论 aspir可抑制ald诱导的CF增殖，其作用机制可能与下调TGF-β-Smad 2、3、4的蛋白水平相关。
목적：탐토아사필림（aspirin，aspi）대철고동（ald）유도적신생대서심기성섬유세포（cardiac fibroblasts， CF）증식적영향급가능궤제。방법0．08％이매소화1～3 d SD신생대서，차속첩벽법순화SD신생대서원대CF。정식실험채용배양3～5대CF，분위4조，공백대조조：무소우혈청적고당DMEM；철고동조（ ald）：무소우혈청적고당DMEM＋1×10－8 mol／L ald；아사필림＋철고동조（ aspi＋ald）：무소우혈청적고당DMEM＋1×10－8 mol／L ald ＋1．11×10－6 mol／L aspi；라내지＋철고동조（ spiro ＋ald ）：무소우혈청적고당 DMEM ＋1×10－8 mol／L ald＋1×10－6 mol／L spiro。 HE염색법관찰각조CF형태학변화；사담서염（ MTT）비색법검측세포증식；Western인적분석TGF-β-Smad 2、3、4적단백표체。결과 HE염색결과현시，여대조조상비，ald가사세포증생활약，분렬상세포현저증다（P＜0．01）；여ald상비，Aspi＋ald조급spiro＋ald조균가사세포분렬현저감소（P＜0．01）。 MTT법검측세포증식결과제시，여대조조상비，ald능구명현증가CF대MTT적대사솔，MTT치명현승고，여대조조상비차이현저（P＜0．01）；여 ald 조상비，aspi ＋ald 조、spiro ＋ald 조균가사CF 대MTT 대사솔감소，MTT치현저강저（P＜0．01）。 Western인적검측결과현시，이ald자격CF후，TGF-β-Smad 2、Smad 3급Smad 4단백수평명현증고，이aspi급spiro간예후，CF중TGF-β-Smad 2、Smad 3급Smad 4단백수평표체교ald조명현강저，균유현저성차이（ P＜0．05）。결론 aspir가억제ald유도적CF증식，기작용궤제가능여하조TGF-β-Smad 2、3、4적단백수평상관。
Objective To investigate the effects of aspirin(aspi) on rat cardiac fibroblasts (CFs) proliferation induced by aldosterone(ald) and the underlying molecular mechanisms .Methods Primary CFs from 1-3 day neonatal rats were digested by 0.08%trypsin and then purified by differential adhesion .The rats were divided into four groups:control group, DMEM medium ( free calf serum ) , ald group [ DMEM medium ( free calf serum ) +ald 1 ×10 -8 mol/L ] , aspi group [DMEM medium (free calf serum)+ald 1 ×10 -8 mol/L+aspi 1.11 ×10 -6 mol/L] and spiro group [DMEM medium (free calf serum)+ald 1 ×10 -8 mol/L +spiro 1 ×10 -6 mol/L].The morphology of CFs was assayed by HE staining methods .MTT Methods were used to measure cell proliferation .Western blotting was used to determine protein expression of TGF-β-Smad 2,3,4.Results HE Staining results showed that compared with the control group , ald activated cell proliferation and increased the cell division phase significantly (P<0.01).Compared with ald group, aspi+ald as well as spiro+ald could reduce cell division significantly ( P<0 .05 ) .MTT assay showed that compared with control group , ald could significantly improve the metabolism of MTT in CF (P <0.01).Compared with ald group, aspi +ald as well as spiro+ald could reduce the metabolism of MTT (P<0.01).Western blotting revealed that the expression levels of TGF-β-Smad 2, 3, 4 in CF were significantly increased by the stimulation of ald but were significantly reduced in aspi +ald and spiro+ald groups compared with ald group (P<0.01).Conclusion Aspi can inhibit the proliferation of CFs induced by ald,possibly by down-regulating the expression of Smad 2, Smad3 and Smad4.