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山东大学学报(医学版) ›› 2013, Vol. 51 ›› Issue (3): 1-5.

• 基础医学 •    下一篇

NAD对AngII诱导的心肌成纤维细胞I型胶原mRNA表达的影响

赵立祥,吴新宁,王希,李娜,陈桐帅,刘军妮,卜培莉   

  1. 山东大学齐鲁医院心内科, 济南 250012
  • 收稿日期:2012-11-16 出版日期:2013-03-10 发布日期:2013-03-10
  • 通讯作者: 卜培莉(1962- ) ,女,博士,主任医师,主要从事心血管重构和高血压病研究。E-mail:bupeili@gmail.com
  • 作者简介:赵立祥(1984- ) ,男,硕士研究生,主要从事心血管重构的研究。E-mail:zhaolixiang841017@126.com
  • 基金资助:

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

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

ZHAO Li-xiang, WU Xin-ning, WANG Xi, LI Na, CHEN Tong-shuai, LIU Jun-ni, BU Pei-li   

  1. Department of Cardiology, Qilu Hospital of Shandong University, Jinan 250012
  • Received:2012-11-16 Online:2013-03-10 Published:2013-03-10

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(1mol/L)+NAD(250μmol/L)组。刺激24h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。

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

Abstract:

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.

Key words: Nicotinamide adenine dinucleotide; Collagen I; Angiotensin II; Fibrosis; Cardiac remodeling; SIRT1

中图分类号: 

  • R542.2
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