丙泊酚通过抑制小鼠海马神经元突触释放组织纤溶酶原激活物引起神经毒性损伤

Propofol Induces Neurotoxicity by Inhibiting tPA Release of Neuronal Synapse in Mouse Hippocampus

目的:明确丙泊酚对体外培养的发育期新生小鼠海马神经元释放组织纤溶酶原激活物(tPA)的影响,并进一步探讨加入外源性tPA能否逆转丙泊酚对发育神经元的毒性损伤作用。方法:将体外培养的发育期小鼠海马神经元分为4组:对照组(C组)、丙泊酚组(Pro组)、tPA组和丙泊酚+tPA组(Pro+tPA组)。C组不做任何处理;Pro组在培养液中加入丙泊酚,终浓度为5、10、30μmol/L,继续孵育不同的时间(1 h、2 h、3 h、6 h);tPA组在培养液中加入tPA,终浓度为1μmol/L,继续孵育6 h;Pro+tPA组在在培养液中同时加入tPA(终浓度1μmol/L)及丙泊酚(终浓度10μmol/L),继续孵育6 h。检测各组神经元凋亡率及凋亡指标actived-caspase-3蛋白的表达情况。结果:Pro组神经元的凋亡率及actived-caspase-3蛋白的表达水平较C组显著上调(P<0.05)。Pro组tPA水平显著低于C组(P<0.05)。Pro+tPA组的神经元的凋亡率及actived-caspase-3蛋白的表达水平显著低于Pro组(P<0.05)。结论:丙泊酚可通过抑制神经元释放tPA而对体外培养的新生小鼠发育期海马神经元产生毒性损伤作用,外源性加入tPA可部分逆转丙泊酚的神经毒性作用。

Objective：To explore the effect of propofol on the tPA release of hippocampal neurons in developing mice,so as to investigate whether exogenous tPA could reverse the propofol-induced neurotoxicity on developing neurons.Methods：The hippocampal neurons of developing mice cultured in vitro were divided into 4 groups（n=20 each）：control group（group C）,propofol group（group Pro）,group tPA and propofol plus tPA group（group Pro＋tPA）.There was no specific treatment in group C.In group Pro,propofol was added to the culture media with the final concentration of 5,10,30μmol/L,respectively,and the cells were then incubated for 1 h,2 h,3 h,6 h,respectively.In group tPA,tPA was added to the culture media with the final concentration of 1μmol/L,and the cells were then incubated for 6 h.In group Pro＋tPA,both propofol and tPA were added to the culture with the final concentration of 10μmol/L and 1μmol/L,respectively,and the cells were then incubated for6 h.The neuron apoptosis rate and the expression level of actived-caspase-3 protein were detected in each group.Results：Compared with that in group C,the apoptosis rate and the expression level of actived-caspase-3 protein in group Pro significantly increased（P〈0.05）.The tPA level in group Pro was significantly higher than that in group C（P〈0.05）.Compared with that in group Pro,the apoptosis rate and the expression level of actived-caspase-3 protein in group Pro＋tPA significantly decreased（P〈0.05）.Conclusions：Propofol induces neurotoxicity on hippocampal neurons of developing mice cultured in vitro by inhibiting neuronal tPA release.And exogenous tPA could partially reverse propofol-induced neurotoxicity.