中华临床医师杂志(电子版)
中華臨床醫師雜誌(電子版)
중화림상의사잡지(전자판)
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2013年
11期
4878-4882
,共5页
高君%魏璨%陈爱东%李梅秀%邵洪江%彭雪%柳磊%徐长庆%李鸿珠
高君%魏璨%陳愛東%李梅秀%邵洪江%彭雪%柳磊%徐長慶%李鴻珠
고군%위찬%진애동%리매수%소홍강%팽설%류뢰%서장경%리홍주
受体,多巴胺%细胞凋亡%再灌注损伤%MAPK通路
受體,多巴胺%細胞凋亡%再灌註損傷%MAPK通路
수체,다파알%세포조망%재관주손상%MAPK통로
Receptors,Dopamine%Apoptosis%Reperfusion injury%MAPK pathways
目的观察2类多巴胺受体( DR2)激活对乳鼠心肌细胞凋亡的影响及其与MAPK通路的关系。方法通过原代培养乳鼠心肌细胞缺氧-复氧模拟缺血-再灌注(ischemia/reperfusion,I/R)损伤模型。心肌细胞随机分为4组:正常对照组(Control,C group),缺血-再灌注组(I/R group),10μmol/L Bromocriptine (DR2激动剂)组(Bro group),10μmol/L Haloperidol(DR2抑制剂)组(Hal group)。 Hoechst33342染色观察细胞凋亡情况;免疫荧光检测心肌细胞促凋亡因子( caspase-3、caspase-9)和抑凋亡因子( Bcl-2)蛋白的表达;Western blot方法检测MAPK通路相关因子p-ERK、p-p38及p-JNK活性变化。结果与Control组相比,I/R组凋亡细胞增加,p-p38、p-JNK、促凋亡因子及抑凋亡因子表达均增加,唯有p-ERK蛋白表达减少;与I/R组比较,Bro可减轻心肌细胞凋亡,下调促凋亡因子的蛋白表达和p-p38、p-JNK的活性,上调抑凋亡因子的蛋白表达和p-ERK活性;Hal则对上述指标影响不明显。结论 DR2的激活可抑制乳鼠心肌细胞凋亡,其机制与MAPK通路相关。
目的觀察2類多巴胺受體( DR2)激活對乳鼠心肌細胞凋亡的影響及其與MAPK通路的關繫。方法通過原代培養乳鼠心肌細胞缺氧-複氧模擬缺血-再灌註(ischemia/reperfusion,I/R)損傷模型。心肌細胞隨機分為4組:正常對照組(Control,C group),缺血-再灌註組(I/R group),10μmol/L Bromocriptine (DR2激動劑)組(Bro group),10μmol/L Haloperidol(DR2抑製劑)組(Hal group)。 Hoechst33342染色觀察細胞凋亡情況;免疫熒光檢測心肌細胞促凋亡因子( caspase-3、caspase-9)和抑凋亡因子( Bcl-2)蛋白的錶達;Western blot方法檢測MAPK通路相關因子p-ERK、p-p38及p-JNK活性變化。結果與Control組相比,I/R組凋亡細胞增加,p-p38、p-JNK、促凋亡因子及抑凋亡因子錶達均增加,唯有p-ERK蛋白錶達減少;與I/R組比較,Bro可減輕心肌細胞凋亡,下調促凋亡因子的蛋白錶達和p-p38、p-JNK的活性,上調抑凋亡因子的蛋白錶達和p-ERK活性;Hal則對上述指標影響不明顯。結論 DR2的激活可抑製乳鼠心肌細胞凋亡,其機製與MAPK通路相關。
목적관찰2류다파알수체( DR2)격활대유서심기세포조망적영향급기여MAPK통로적관계。방법통과원대배양유서심기세포결양-복양모의결혈-재관주(ischemia/reperfusion,I/R)손상모형。심기세포수궤분위4조:정상대조조(Control,C group),결혈-재관주조(I/R group),10μmol/L Bromocriptine (DR2격동제)조(Bro group),10μmol/L Haloperidol(DR2억제제)조(Hal group)。 Hoechst33342염색관찰세포조망정황;면역형광검측심기세포촉조망인자( caspase-3、caspase-9)화억조망인자( Bcl-2)단백적표체;Western blot방법검측MAPK통로상관인자p-ERK、p-p38급p-JNK활성변화。결과여Control조상비,I/R조조망세포증가,p-p38、p-JNK、촉조망인자급억조망인자표체균증가,유유p-ERK단백표체감소;여I/R조비교,Bro가감경심기세포조망,하조촉조망인자적단백표체화p-p38、p-JNK적활성,상조억조망인자적단백표체화p-ERK활성;Hal칙대상술지표영향불명현。결론 DR2적격활가억제유서심기세포조망,기궤제여MAPK통로상관。
Objective To observe the effect of DR 2 activation on cell apoptosis in the neonatal rat cardiomyocytes ,and to explore the relationship with MAPK pathways .Methods The ischemia/reperfusion ( I/R ) injury model was established in primarily cultured neonatal rat cardiomyocytes by anoxia -reoxygenation ,and randomly assigned:Control,I/R,Bro and Hal groups.The cell apoptosis was detected using Hoechst 33342 dye.The expression of protein of caspase-3, caspase-9 and Bcl-2 was detected by immunofluorescence .The activity of MAPK-related factors as p-ERK, p-p38 and p-JNK was detected by Western blot .Results Compared with the control group , apoptosis rate ,the activity of p-p38 and p-JNK and expression of pro-apoptotic factors and anti-apoptotic factors were increased,but p-ERK activity was decreased .Compared with the I/R group,all index above-mentioned were down-regulated or reversed in Bro-group,and had no obvious differences in Hal group .Conclusion DR2 activation inhibit apoptosis of the neonatal rat cardiomyocytes apoptosis ,which mechanism is related to MAPK pathways .