摘要
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]亮氨酸的摄人情况;免疫组化染色分析心肌细胞的横截面积;CM-H_2DCFDA检测细胞活性氧（ROS）的生成情况;试剂盒检测细胞内H_2O_2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50μmol/L浓度作用最强（P〈0.05）;（2）在Apelin 50μmol/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 50μmol/L可减少胞内H_2O_2水平,且采用ATZ预处理后可抑制Apelin降低胞内H_2O_2的作用（P〈0.05）;（4）Apelin 50μmol/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。
Objective To explore the inhibition effect of Apelin on the oxidative stress related cardiac hypertrophy and the possibly mechanism. Methods The 5-I-IT 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 0-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-H2 DCFDA. 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 μmol/L （ P 〈 0.05） ; （2） The concentration of 50 μmol/L Apelin could reduce the level of intra- cellular 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 H2O2, 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 ）. Con- clusion It demonstrated that the Apelin inhibit the oxidative stress related cardiac hypertrophy, with the possibly mechanism was the activation of catalase.
出处
《疑难病杂志》
CAS
2012年第7期527-530,共4页
Chinese Journal of Difficult and Complicated Cases
基金
国家自然科学基金项目(No.30500657)
