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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。
Objective To investigate the effects of type 2 dopamine receptor (DR2) activation on neonatal rat cardiomyocyte apoptosis and its relationship with MAPK pathway. Methods Primary cultured neonatal rat cardiomyocytes were subjected to hypoxia - reoxygenation simulated ischemia - reperfusion (I / R) injury model. Cardiomyocytes were randomly divided into 4 groups: Control, C group, I / R group, 10μmol / L Bromocriptine (Bro group), 10μmol / L Haloperidol (DR2 inhibitor) group (Hal group). Hoechst33342 staining was used to observe the cell apoptosis. The expression of caspase-3 and Bcl-2 protein was detected by immunofluorescence. The expression of MAPK pathway-related factor p- ERK, p-p38 and p-JNK activity changes. Results Compared with Control group, the number of apoptotic cells in I / R group increased and the expression of p-p38, p-JNK, pro-apoptotic factor and pro-apoptotic factor increased, but p-ERK protein expression decreased. Bro reduced cardiomyocyte apoptosis, down-regulated the expression of pro-apoptotic factor and p-p38, p-JNK, and up-regulated the protein and p-ERK activity of apoptotic factor. Hal also affected the above indexes Not obvious. Conclusion Activation of DR2 can inhibit cardiomyocyte apoptosis in neonatal rats, and its mechanism is related to MAPK pathway.