CAJ | 학술논문

目的评价前列腺素E2受体(EP受体)在前列腺素E2(PCE2)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1 ml)或6孔(每孔2 ml)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48 h;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(EP1及EP2受体拮抗剂,终浓度10μmol/L);GW627368X组(G组)在细胞培养液中加入PGE2(终浓度1μmol/L)和GW627368X(EP4受体拮抗剂,终浓度10 μmol/L).孵育48 h后采用免疫荧光观察心肌细胞形态,Image J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ANP mRNA及BNP mRNA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PGE2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ANP mRNA及BNP mRNA表达下调(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EP1和EP2受体无关.
목적 평개전렬선소E2수체(EP수체)재전렬선소E2(PCE2)유도H9c2심기세포비대중적작용.방법 배양H9c2심기세포,이4×104개/ml적밀도접충우배양병(매병3ml)、24공(매공1 ml)혹6공(매공2 ml)배양판.채용수궤수자표법,장세포수궤분위4조(n=24):공백대조조(C조)불여임하처리,계속배양48 h;PGE2조재세포배양액중가입PGE2(종농도1μmol/L);AH6809조(A조)재세포배양액중가입PGE2(종농도1μmol/L)화AH6809(EP1급EP2수체길항제,종농도10μmol/L);GW627368X조(G조)재세포배양액중가입PGE2(종농도1μmol/L)화GW627368X(EP4수체길항제,종농도10 μmol/L).부육48 h후채용면역형광관찰심기세포형태,Image J의학도상분석계통측량심기세포직경,BCA법검측심기세포총단백함량,RT-PCR법측정포장ANP mRNA급BNP mRNA적표체수평.결과 여C조비교,PGE2조、A조화G조심기세포총단백함량화심기세포직경증가,포장ANPmRNA급BNPmRNA표체상조(P＜0.05).여PGE2조비교,G조심기세포총단백함량화심기세포직경강저,포장ANP mRNA급BNP mRNA표체하조(P＜0.05),A조상술각지표차이무통계학의의(P＞0.05).결론 EP4수체개도료PGE2유도적심기세포비대효응,이해효응여EP1화EP2수체무관.
Objective To evaluate the role of prostaglandin E2 (EP) receptors in H9c2 cardiomyocyte hypertrophy induced by prostaglandin E2 (PGE2).Methods Primary cultured H9c2 cardiomyocytes 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 (EP1 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 1 μmol/L) was added to the culture medium in PGE2 group.PGE2 (final concentration 1 μmol/L) and A H6809 (final concentration 10 μmol/L) were added to the culture medium in group A.PGE2 (final concentration 1 μmol/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 expression 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 diameter were significantly decreased,and the expression of ANP mRNA and BNP mRNA in the cytoplasm was downregulated 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.