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[目的]观察急性缺血心肌心脏型脂肪酸结合蛋白(H-FABP)基因及蛋白表达的动态变化,揭示H-FABP在心肌代谢异常中的作用。[方法]健康SD雄性大鼠随机分为sham组和5个实验组(A1,A2,A3,A4和A5分别为心肌缺血1,2,4,6h和12h);应用HE染色观察各实验组心肌缺血后病理形态学改变,用RT-PCR和WesternBlot的方法检测心肌H-FABP基因和蛋白表达的变化,用一步提取比色法测定血清游离脂肪酸(FFA)含量。[结果](1)缺血2、4h组心肌细胞明显肿胀,有部分形成了“收缩带”;缺血6h和12h心肌细胞边界不清,局灶性凝集坏死,缺血12h组有大量炎性细胞浸润。(2)RT-PCR结果显示:A2、A3、A4、A5组与sham组比较心肌H-FABP基因明显下降(P﹤0.05),尤其A3、A4组降低分别约29%,30%(P﹤0.01)。(3)WesternBlot检测:A2、A3、A4、A5组与sham组比较心肌H-FABP蛋白均表达下降,尤其A4组降低约41%(P﹤0.01)。(4)FFA测定:A2、A3、A4、A5组与sham组比较大鼠血清的FFA含量均有不同程度的升高,尤以缺血2、4h组为甚(P﹤0.01),分别升高约2.01倍和2.72倍。[结论]急性心肌缺血后心脏型脂肪酸结合蛋白基因及蛋白表达明显下降,可能参与心肌细胞代谢紊乱,是导致局部心肌损伤的机制之一。
[Objective] To observe the dynamic changes of H-FABP gene and protein expression in acute ischemic myocardium and to reveal the role of H-FABP in abnormal myocardial metabolism. [Methods] The healthy SD male rats were randomly divided into sham group and five experimental groups (1, 2, 4, 6h and 12h after A1, A2, A3, A4 and A5, respectively) The changes of myocardial H-FABP gene and protein expression were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The level of serum free fatty acid (FFA) was determined by one-step extraction and colorimetry. [Results] (1) Myocardial cells in ischemic group were significantly swollen in 2,4h group, some formed “systolic zone”, and the border of myocardial cells were unclear at 6h and 12h after ischemia, focal coagulation and necrosis, A large number of inflammatory cell infiltration. (2) The results of RT-PCR showed that the H-FABP gene expression of myocardium in A2, A3, A4 and A5 groups was significantly decreased compared with sham group (P <0.05), especially in A3 and A4 groups decreased by 29% and 30% 0.01). (3) Western blot analysis showed that the expression of H-FABP protein decreased in A2, A3, A4 and A5 groups compared with sham group, especially in A4 group (about 41%, P <0.01). (4) The levels of FFA in serum of rats in groups A2, A3, A4 and A5 were significantly higher than those in sham group (P <0.01) About 2.01 times and 2.72 times higher. [Conclusion] The gene and protein expression of cardiac fatty acid-binding protein in acute myocardial ischemia decreased significantly, which may be involved in the metabolic disorder of myocardial cells, which is one of the mechanisms leading to local myocardial injury.