丙泊酚诱导血红素氧化酶-1表达抑制氧糖剥夺大鼠海马神经元凋亡及机制研究

Propofol prevents hippocamp neurons from oxygen-glucose deprivation injury through inducing HO-1 protein expression

目的研究丙泊酚诱导血红素氧化酶-1(HO-1)蛋白表达抑制氧糖剥夺海马神经元凋亡的机制。方法将培养7d的新生(出生24～48h)SD大鼠海马神经元随机均分为五组:C1组,海马神经元全量换液后再继续培养24h;C2组,海马神经元接受50μmol/L丙泊酚处理1h后,再继续培养24h;D组,海马神经元进行缺糖缺氧培养1h后复糖复氧,再继续培养24h;P组,海马神经元缺糖缺氧的同时接受50μmol/L丙泊酚处理;Z组,在海马神经元进行缺糖缺氧的同时加入锌原卟啉(ZnpplX)使其终浓度为10μmol/L后处理同P组。每组分别取12孔海马神经元,用Hoechst33342染色法检测细胞凋亡,用免疫细胞化学染色法检测HO-1蛋白和Bcl-2蛋白的表达。结果与C1组比较,C2、D及P组海马神经元HO-l和Bcl-2蛋白的表达均增加,D、P及Z组凋亡率显著升高(P<0.05或P<0.01)。与D组比较,P组海马神经元HO-l和Bcl-2蛋白表达均增加,凋亡率下降(P<0.01)。与P组比较,Z组海马神经元HO-l和Bcl-2蛋白的表达均降低,凋亡率则明显升高(P<0.01)。结论丙泊酚通过诱导氧糖剥夺海马神经元表达HO-1,进而上调Bcl-2,从而抑制氧糖剥夺海马神经元的凋亡,可能是丙泊酚神经保护作用的机制之一。

Objective To investigate the neuroprotective elects of propofol on oxygen-glucose deprivation injury to primary culture rats hippocamal neurons. Methods Newborn（24-48 h）SD rats hippocampus neurons were cultured 7 days. On the 7th day the cultured hippocampus neurons were randomly divided into five group： Group C1 were normally incubated another 24 h after replacing all culture solution. Group C2 were incubated another 24 h after hippocampus neurons were cultured with 50μmol/L propofol for 1 h. Group D were normally incubated another 24 h after hippocampus neurons were carried out oxygen glucose deprivation （OGD） 1 h. Group P were normally incubated another 24 h after hippocampus neurons were carried out OGD and meanwhile were cultured with 50 μmol/I, propofol 1 h. Group Z cells were incubated normally for another 24 h after hippocampus neurons were carried out OGD 1 hour and meanwhile were added 50μmol/L propofol and 10 μmol/1. ZnpplX. After normal culture 24 h, apoptosis rate was detected by Hoechst33342 staining technique and fluorescence microscopy, HO-1 and Bcl-2 protein expression was detected by immunocytochemical technique. Results Compared with group C1, the expression of HO-1 and Bcl-2 protein were both increased in groups C2, D and P, apoptosis rates were increased in groups D,P and Z（P〈0.05 or P 〈0.01）. Compared with group D, the expression of HO-1 and Bcl-2 protein were both increased and apoptosis rates were decreased in group P（P〈0.01）. Compared with group P, the expression of HO- 1 and Bcl-2 protein were both decreased and apoptosis rates were increased in group Z（P〈0. 01）. Conclusion Propofol inducing HO-1 expression, protects neurons against oxygen-glucose deprivation injury through up-regulating Bcl-2.