硫化氢下调TGF-β_1/Smad2/3通路对大鼠心肌细胞胶原合成的影响  

Effect of hydrogen sulfide on collagen synthesis in rat cardiomyocytes through down regulation of TGF-β_1/Smad2/3 signaling pathway

作　　者：刘百川[1] 孙莉莉[1] 王海英[1] 孙鲁静 赵晓东[2] 赵斌[1] 

机构地区：[1]北京积水潭医院急诊科,北京100035 [2]解放军总医院第一附属医院急救部,北京100048

出　　处：《解放军医学杂志》2014年第9期681-684,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(31130030)~~

摘　　要：目的探讨气体分子硫化氢(H2S)对大鼠心肌细胞胶原合成的调节作用及可能机制。方法将大鼠心肌细胞H9C2随机分为4组:正常对照组(加入DMEM培养基)、硫氢化钠(NaSH)组(加入终浓度为2×10–4mol/L的H2S供体NaHS培养)、TGF-β组(加入终浓度为10ng/ml TGF-β1培养)、TGF-β+NaHS组(用含2×10–4mol/L NaHS的DMEM培养基培养30min,再加入10ng/ml TGF-β1培养)。培养1h后,采用Western blotting检测心肌细胞p-Smad2和p-Smad3蛋白表达水平,培养72h后采用免疫荧光染色法观察心肌细胞的Ⅰ型胶原蛋白的表达。结果与正常对照组比较,NaHS组心肌细胞p-Smad2和pSmad3蛋白表达水平无明显差异,而TGF-β组心肌细胞p-Smad2和p-Smad3蛋白表达明显增多,且Ⅰ型胶原蛋白表达明显增强。与TGF-β组比较,TGF-β+NaHS组心肌细胞p-Smad2和p-Smad3蛋白表达明显下降,Ⅰ型胶原蛋白表达明显减弱。结论 H2S可抑制心肌细胞胶原合成,其机制可能与阻断TGF-β1/Smad2/3信号转导通路有关。Objective To explore the role of hydrogen sulfide （H2S）, a novel gaseous transmitter, in the regulation of rat myocardial collagen synthesis and the possible mechanisms thereof. Methods Rat myocardial cell line （H9C2） was divided into four groups： normal control group （DMEM medium was used）, NariS group （NariS at concentration of 2 × 10^-4mol/L was used）, TGF-β group （TGF-β1 at a final concentration of 10ng/ml was added） and TGF-β＋NaHS group （cultured with 2 × 10^-4 mol/L Naris for 30min, and then 10ng/ml TGF-β1 was added）. After cultivation for lh, p-Smad2 and p-Smad3 expressions in cardiomyocytes were assessed with Western blotting. After cultivation for 72h, collagen type I expression was determined by immunofluorescence and confocal microscopy. Results Compared with control group, the expressions of p-Smad2, p-Smad3 and collagen type I significantly increased in TGF-β group, but there was no significant difference in NariS group. Compared with TGF-β group, the expressions of p-Smad2, p-Smad3 and collagen type I stimulated by TGF-β1 significantly decreased in TGF-β＋NaHS group. Conclusions H2S can prevent collagen synthesis in rat cardiomyocytes. The mechanism might involve the inhibition of TGF-β1/Smad2/3 signaling pathway.

关 键 词：硫化氢 肌细胞 心脏 转化生长因子β1 SMAD信号通路 

分 类 号：Q257[生物学—细胞生物学]