Objective To explore the inhibition effect of Apelin on the oxidative stress related cardiac hypertrophy and the possibly mechanism. Methods The 5-HT was applied to induce the cardiac hypertrophic cells with neonatal rat cardiac myocytes, which was divided into control group, Apelin activation group, inhibition group ( DETC or ATZ ), and combination group. The β-liquid scintillation counter was used to detect the[ 3H ] leucine incorporation and imunohistochemical staining was used to analyze the cell surface area. The intracellular ROS was detected by CM-H2DCFDA. The kits were used to detect the intracellular H2O2 levels, lipid peroxide ( LPO ) levels, catalase ( CAT ) and glutathione peroxidase ( GPx ) activity. Besides, the level of CAT mRNA and GPx mRNA levels were detected by RT-PCR. Results ( 1 ) Compared with control group, the Apelin reduced the[ 3H ] leucine incorporation and cell surface of cardiac hypertrophic cells in a dose dependent manner with the strongest effect at 50 μmnol/L ( P < 0. 05 ); (2) The concentration of 50 μmol/L Apelin could reduce the level of intracellular ROS ( P <0.05 ), but the pretreatment with DETC significantly inhibited the effects of Apelin ( P <0. 05 ); (3) The concentration of 50 μmol/L Apelin could reduce the level of intracellular H2 O2, and the pretreatment with ATZ inhibited the effects of Apelin ( P < 0. 05 ); (4) The concentration of 50 μmol/L Apelin treatment could result in the improvement of the CAT activity, mRNA levels increased, while the LPO, GPx activity and mRNA levels did not have difference ( P > 0.05 ), while the pretreatment with ATZ significantly inhibited the CAT activity, mRNA levels increase ( P < 0. 05 ). Conclusion It demonstrated that the Apelin inhibit the oxidative stress related cardiac hypertrophy, with the possibly mechanism was the activation of catalase.%目的 探讨Apelin对于氧化应激相关心肌肥大的抑制作用及可能的作用机制.方法 采用5-羟色胺(5-HT)刺激新生大鼠心肌细胞诱导心肌肥大细胞,而后分成对照组、Apelin处理组、抑制剂(DETC或ATZ)处理组、联合处理组;β-液体闪烁计数仪检测[3H]亮氨酸的摄入情况;免疫组化染色分析心肌细胞的横截面积;CM-H2DCFDA检测细胞活性氧(ROS)的生成情况;试剂盒检测细胞内H2O2水平、过氧化氢脂质(LPO)水平、过氧化氢酶(CAT)活性以及谷胱甘肽过氧化物酶(GPx)活性; RT-PCR检测CAT mRNA以及GPx mRNA水平.结果 (1)与对照组相比,Apelin呈浓度依赖性地降低心肌肥大细胞[3H]亮氨酸的摄入以及细胞横裁面积,且以50 μmol/L浓度作用最强(P<0.05);(2)在Apelin 50 μmol/L培养下,可减少细胞内ROS水平(P < 0.05),但采用DETC预处理后能够显著抑制Apelin降低ROS合成的作用(P<0.05);(3)Apelin 50 μmol/L可减少胞内H2O2水平,且采用ATZ预处理后可抑制Apelin降低胞内H2O2的作用(P<0.05);(4)Apelin 50 μmol/L处理后的CAT活性、mRNA水平升高,而LPO、GPx的活性和mRNA水平没有差异(P>0.05),采用ATZ预处理后可显著抑制过CAT活性、mRNA水平的升高(P<0.05).结论 Apelin可抑制氧化应激相关的心肌肥大,可能机制是激活过氧化氢酶.
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