NAD对AngⅡ诱导的心肌成纤维细胞Ⅰ型胶原mRNA表达的影响  

Effects of NAD on the mRNA expression of collagen type Ⅰ induced by Angiotensin Ⅱ in cardiac fibroblasts

作　　者：赵立祥[1] 吴新宁[1] 王希[1] 李娜[1] 陈桐帅 刘军妮[1] 卜培莉[1] 

机构地区：[1]山东大学齐鲁医院心内科, 济南250012

出　　处：《山东大学学报（医学版）》2013年第3期1-5,共5页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(81070076,81170135);国家973项目(2012CB722406)

摘　　要：目的探讨烟酰胺腺嘌呤二核苷酸(NAD)对血管紧张素II(AngII)诱导的大鼠心肌成纤维细胞I型胶原mRNA表达的作用。方法提取新生Wistar大乳鼠原代心肌成纤维细胞,传代培养,采用2～4代细胞。实验分为空白对照组,AngII(0.01、0.1、1μmol/L)组,NAD(250μmol/L)组,AngII(1μmol/L)+NAD(250μmol/L)组,FAM组,Sirt1-siRNA+AngII(1μmol/L)组,Sirt1-siRNA组,Sirt1-siRNA+NAD(250μmol/L)组,Sirt1-siRNA+Ang II(1 mol/L)+NAD(250μmol/L)组。刺激24 h后,提取总RNA,Real time RT-PCR法分别检测SIRT1和I型胶原mRNA的表达。结果心肌成纤维细胞I型胶原mRNA的表达随着AngⅡ浓度升高明显增加(P<0.05),呈浓度依赖性。与空白对照组比较,NAD(250μmol/L)组SIRT1 mRNA表达增高(P<0.05)。与AngⅡ(1μmol/L)组比较,AngII(1μmol/L)+NAD(250μmol/L)组I型胶原mRNA的表达明显减少(P<0.01)。相对于FAM组,Sirt1-siRNA转染后各组SIRT1 mRNA的表达减少(P<0.05),而心肌成纤维细胞I型胶原mRNA表达增多(P<0.05)。结论 NAD可以抑制心肌成纤维细胞I型胶原mRNA的表达,SIRT1影响I型胶原mRNA的表达,它们对Ang II诱导的心肌纤维化有保护作用。Objective To explore the effect of NAD on the mRNA expression of collagen type I induced by angiotensin Ⅱ in cardiac fibroblasts in vitro.Methods Neonatal rat cardiac fibroblasts were isolated from wistar rats and then cultured. The generation 2-4 were used for the experiment and treated as follows： control group, Ang Ⅱ （0.01, 0.1, 1μmol/L） group,NAD （250μmol/L ）group, Ang II （1μmol/L）＋ NAD （250μmol/L ）group, FAM group, Sirt1-siRNA＋AngⅡ （1μmol/L） group, Sirt1-siRNA group, Sirt1-siRNA＋NAD（250μmol/L）group and Sirt1-siRNA＋ Ang Ⅱ （1μmol/L）＋NAD（250μmol/L）group. 24 hours later, the mRNA expressions of SIRT1 and collagen I were measured by real time reverse transcription-polymerase chain reaction（Real time RT-PCR）. Results After treatment with Ang Ⅱ for 24 hours,the mRNA expression of collagen I was enhanced in a Ang II dose-dependent manner（P〈0.05）. In contrast with control group, the mRNA expression of SIRT1 was enhanced after being treated with NAD（250μmol/L）（P〈0.05） and the mRNA expression of collagen I in AngII （1μmol/L）＋ NAD （250μmol/L） group was attenuated compared with Ang Ⅱ （1μmol/L） group （P〈0.01）. Compared with FAM group, SIRT1 mRNA expression was attenuated（P〈0.05） and collagen I mRNA expression was increased （P〈0.05） in the other groups.Conclusion The results suggest that NAD and SIRT1 may be beneficial to cardiac fibrosis by attenuating the mRNA expression of collagen type I induced by Ang Ⅱ in cardiac fibroblatsts.

关 键 词：烟酰胺腺嘌呤二核苷酸 I型胶原 血管紧张素II 纤维化 心肌重构 SIRT1 

分 类 号：R542.2[医药卫生—心血管疾病]