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目的研究Raf-1 siRNA重组腺病毒载体对大鼠心肌肥厚的作用并探讨其机制。方法构建Raf-1腺病毒siRNA载体,大鼠随机分为正常对照组、心肌肥厚组、无关siRNA组、Raf-1 siRNA干预组,皮下注射异丙肾上腺素(ISO)制备大鼠心肌肥厚模型,无关siRNA组及Raf-1 siRNA干预组注射ISO后,分别经心包腔注射重组腺病毒p Ad-siCon及p Ad-siRaf-1,4周后染色测定全心重指数(HWI)、心肌细胞横截面积(CSA)、心肌间质胶原容积分数(CVF)及血管周围胶原面积(PVCA),western blot检测心肌Raf-1、NF-κB及ERK1/2蛋白表达,RT-PCR检测TNF-α、MCP-1、TGF-β1 mRNA表达。结果与心肌肥厚组相比,Raf-1 siRNA干预组可降低大鼠HWI、CSA、CVF及PVCA,下调Raf-1、NF-κB、ERK1/2蛋白及TNF-α、MCP-1、TGF-β1 mRNA表达;无关siRNA组与心肌肥厚组相比无明显差异。结论 Raf-1 siRNA重组腺病毒载体能减轻大鼠心肌肥厚,其作用可能与抑制Raf-1/NF-κB、Raf-1/ERK1/2信号转导及下调肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)、转化生长因子-β(TGF-β)表达相关。
Objective To investigate the effect of Raf-1 siRNA recombinant adenovirus vector on cardiac hypertrophy in rats and its mechanism. Methods The Raf-1 adenovirus siRNA vector was constructed. The rats were randomly divided into normal control group, cardiac hypertrophy group, unrelated siRNA group, Raf-1 siRNA intervention group and subcutaneous injection of isoproterenol (ISO) After injection of ISO, unrelated siRNA group and Raf-1 siRNA intervention group were injected intrapericardially with recombinant adenovirus p Ad-siCon and p Ad-siRaf-1, 4 weeks after the whole heart weight index (HWI), cardiomyocyte horizontal (CSA), myocardial interstitial collagen volume fraction (CVF) and perivascular collagen area (PVCA). The expressions of Raf-1, NF-κB and ERK1 / 2 were detected by Western blot. The expressions of TNF- MCP-1, TGF-β1 mRNA expression. Results Compared with myocardial hypertrophy group, Raf-1 siRNA could decrease HWI, CSA, CVF and PVCA, downregulate Raf-1, NF-κB, ERK1 / 2 and TNF-α, MCP- β1 mRNA expression; no significant difference between siRNA group and cardiac hypertrophy group. Conclusions Raf-1 siRNA recombinant adenovirus vector can attenuate cardiac hypertrophy in rats, which may be related to the inhibition of Raf-1 / NF-κB, Raf-1 / ERK1 / 2 signal transduction and downregulation of tumor necrosis factor-α ), Monocyte chemoattractant protein-1 (MCP-1) and transforming growth factor-β (TGF-β).