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目的:观察多巴胺受体(DR2)激活对乳鼠心肌细胞缺氧/再灌注损伤的影响,并探讨其机制。方法:复制原代培养乳鼠心肌细胞缺氧/再灌注损伤模型,细胞随机分为正常组(Control)、缺氧/再灌注组(H/R)、DR2激动剂组(溴麦角环肽,Bro)、抑制剂组(氟哌啶醇,Hal)。倒置显微镜、透射电镜、流式细胞仪检测细胞凋亡情况;检测细胞培养液乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;RT-PCR和Western blot方法检测心肌细胞促凋亡因子(Cyt C、caspase-3、caspase-8、caspase-9、Fas及Fas-L)及抑凋亡因子(Bc-l 2)mRNA和蛋白表达。结果:与正常组相比,H/R组细胞凋亡率、LDH活性、MDA含量、促凋亡因子及抑凋亡因子表达均增加,唯有SOD活性降低;与H/R组比较,Bro组可减轻或逆转上述指标的变化;Hal组上述指标变化不明显。结论:DR2激活可抑制缺氧/再灌注所致的乳鼠心肌细胞损伤和凋亡,机制与减少氧自由基有关。
Objective: To observe the effect of dopamine receptor (DR2) activation on neonatal rat cardiomyocyte hypoxia / reperfusion injury and to explore its mechanism. Methods: Primary cultured neonatal rat cardiomyocytes were exposed to hypoxia / reperfusion injury. The cells were randomly divided into control group, hypoxia / reperfusion group (H / R), DR2 agonist group (bromocriptine, Bro), inhibitor group (haloperidol, Hal). The apoptosis of cells was detected by inverted microscope, transmission electron microscopy and flow cytometry. The activity of LDH, SOD and the content of malondialdehyde (MDA) The expressions of Cyt C, caspase-3, caspase-8, caspase-9, Fas, FasL and Bcl-2 mRNA and protein in cardiomyocytes were detected by Western blot. Results: Compared with normal group, the apoptosis rate, LDH activity, MDA content, pro-apoptotic factor and anti-apoptotic factor in H / R group were increased, but only SOD activity was decreased. Compared with H / R group, Bro Group can reduce or reverse the changes of the above indicators; Hal group indicators did not change significantly. Conclusion: DR2 activation can inhibit myocardial cell injury and apoptosis induced by hypoxia / reperfusion in neonatal rats. The mechanism is related to the reduction of oxygen free radicals.