摘要
目的:研究纯化的心肌细胞中结缔组织生长因子(CTGF)基因及蛋白的表达,并探讨转化生长因子β1(TGF-β1)对其表达的影响。方法:分离培养乳鼠心肌细胞,48h后用流式分选方法(FACS)分选纯化心肌细胞,纯化的心肌细胞继续培养48h。分为对照组(空白纯化心肌细胞组)和TGF-β1刺激组,用RT-PCR方法测定两组细胞中TGF-β1、CTGF、纤维连接蛋白(FN)mRNA的含量,用Western blotting法检测两组细胞胞浆蛋白中CTGF、FN含量。结果:心肌细胞中有较低水平的CTGF表达,在予以TGF-β1刺激后细胞内CTGFmRNA与蛋白含量分别提高2.49倍(P<0.01)和3.63倍(P<0.01),FN mRNA与蛋白含量分别提高1倍(P<0.01)和4.18倍(P<0.01),而TGF-β1mRNA水平减少34%(P<0.01)。结论:纯化的心肌细胞有CTGF的基础表达,并且TGF-β1可以明显上调其表达,提示心肌细胞主动参与了心肌纤维化过程。
AIM ： To investigate the mRNA and protein expressions of connective tissue growth factor （CTGF） in the cardiomyocytes, and to evaluate the effect of transforming growth factor-β1 （TGF -β1） on it. METHODS： Ventricular cardiomyocytes were isolated from neonatal rats and cultured for 4;8 h, then the cardiomyocytes were sorted by means of fluorescence - activated cell sorting （FACS）. The sorted cells were continuously cultured for 48 h and divided in- to two groups： the control group and the TGF - β1 stimulation group. The mRNA expressions of CTGF, TGF -β1, and fibronectin （FN） were measured by reverse transcription-polymerase chain reaction （RT- PCR）. Meanwhile, the expressions of CTGF and FN in the plasmosin were detected by Western blotting. RESULTS ： There was basal expression of CTGF in the control group. After stimulation with TGF -β1, the mRNA and protein expressions of CTGF were remarkably increased by 2.49 - folds and 3.63 - folds （ P 〈 0.01 ） and the mRNA and protein expressions of FN were increased by I - fold and 4. 18 -folds （P 〈0.01 ） ,respectively, compared with the control group. However, the mRNA expression of TGF -β1 was decreased by 34% （P 〈0.01 ）. CONCLUSION： Some fundamental secretion of CTGF in the cardiomyocytes and TGF -β1 may induce its expression significantly. The cardiomyocytes participate actively in the progress of the myocardial fibrosis.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2008年第6期1060-1064,共5页
Chinese Journal of Pathophysiology
基金
湖北省教育厅重点资助项目(NoD200624006)
关键词
心肌细胞
心肌纤维化
结缔组织生长因子
转化生长因子Β
Cardiomyocytes
Myocardial fibrosis
Connective tissue growth factor
Transforming growth factor beta
