前列腺素E2受体在前列腺素E2诱导H9c2心肌细胞肥大中的作用

Role of prostaglandin E2 receptors in H9c2 cardiomyocyte hypertrophy induced by prostaglandin E2

目的评价前列腺素E2受体（EP受体）在前列腺素E2（PGE2）诱导H9c2心肌细胞肥大中的作用。方法培养H9c2心肌细胞，以4×104个／ml的密度接种于培养瓶（每瓶3ml）、24孔（每孔1ml）或6孔（每孔2ml）培养板。采用随机数字表法，将细胞随机分为4组（n=24）：空白对照组（c组）不予任何处理，继续培养48h；PGE2组在细胞培养液中加入PGE2（终浓度1μmol／L）；AH6809组（A组）在细胞培养液中加入PGE2（终浓度1μmol／L）和AH6809（EPJ及ER受体拮抗剂，终浓度10μmol／L）；GW627368X组（G组）在细胞培养液中加入PGE，（终浓度1μmol／L）和GW627368X（EP4受体拮抗剂，终浓度10μmol／L）。孵育48h后采用免疫荧光观察心肌细胞形态，ImageJ医学图像分析系统测量心肌细胞直径，BCA法检测心肌细胞总蛋白含量，RT-PCR法测定胞浆ANPmRNA及BNPmRNA的表达水平。结果与C组比较，PGE2组、A组和G组心肌细胞总蛋白含量和心肌细胞直径增加，胞浆ANPmRNA及BNPmRNA表达上调（P〈0．05）。与PGE，组比较，G组心肌细胞总蛋白含量和心肌细胞直径降低，胞浆ANPmRNA及BNPmRNA表达F调（P〈0．05），A组上述各指标差异无统计学意义（P〉0．05）。结论EP4受体介导了PGE2诱导的心肌细胞肥大效应，而该效应与EP，和EP2受体无关。

Objective To evaluate the role of prostaglandin E2 （EP） receptors in H9c2 cardiomyocyte hy- pertrophy induced by prostaglandin E2 （ PGE2 ）. Methods Primary cultured H9c2 cardiomyoeytes were seeded in culture flasks （3 ml/flask） or in 24-well plate （1 ml/hole） or 6-well plate （2 ml/hole） with density of 4 × 104/ml. The cells were randomly divided into 4 groups （ n = 24 each） ： control group （group C）, PGE2 group, AH6809 （ EPt and EP2 receptor antagonist） group （group A） and GW627368X （EP4 receptor antagonist） group （group G）. The cells were continuously cultured for 48 h. PGE2 （final concentration 1μmol/L） was added to the culture medi- um in PGEz group. PGE2 （final concentration l μmol/L） and AH6809 （final concentration 10 μmol/L） were added to the culture medium in group A. PGE2 （final concentration 1μmol/L） and GW627368X （final concentration 10 μmol/L） were added to the culture medium. The cells were then cultured for 48 h in groups PGE2 , A and G. Then the cell morphology was observed by using fluorescent microscope. The cell diameter was measured by using the Image J medical image analysis system. Total protein content in the cells was measured with BCA method. The ex- pression of atrial natriuretic peptide （ANP） mRNA and brain natriuretic peptide （BNP） mRNA in the cytoplasm was determined using RT-PCR.Results Compared with group C, the total protein in the cells and cell diameter were significantly increased, and the expression of ANP mRNA and BNP mRNA in the cytoplasm was up-regulated in groups PGE2, A and G （ P 〈 0.05）. Compared with group PGE2, the total protein in the cells and cell diame- ter were significantly decreased, and the expression of ANP mRNA and BNP mRNA in the cytoplasm was down- regulated in group G （P 〈 0.05）, and no significant change was found in the parameters mentioned above in group A （ P 〉 0.05）. Conclusion EP4 receptor mediates H9c2 cardiomyocyte hypertrophy induced by PGE2 and the effect is not related to EP1 and EP2 .