乙醇对原代培养大鼠皮质神经细胞CYP2E1、iNOS和NSE表达的影响

目的:观察乙醇作用后原代培养大鼠皮质神经细胞CYP2E1、诱导型一氧化氮合酶(iNOS)和神经元特异性烯醇化酶(NSE)表达的变化.方法:分别采用0.5、1.5和3.0 g/L乙醇作用于原代培养大鼠皮质神经细胞,对照组细胞以等量生理盐水处理.作用30 min和1、2、4、8、12、24 h后,采用免疫细胞化学染色法检测皮质神经细胞CYP2E1、iNOS和NSE的表达.结果:乙醇可以增强大鼠皮质神经细胞CYP2E1、iNOS的表达,降低NSE的表达,并且对CYP2E1(F剂量=57.123,F时间=12.111,F交互=4.987,P均＜0.001)、iNOS(F剂量=55.081,F时间=10.949,F交互=4.278,P均＜0.001)和NSE(F剂量=69.334,F时间=10.871,F交互=3.252,P均＜0.001)表达的影响均表现出一定的剂量依赖性和时序性.1.5 g/L乙醇作用2 h时3者表达开始变化,12 h时作用最强;3.0 g/L乙醇作用1 h时3者表达开始变化,8 h时作用最强.结论:乙醇能诱导大鼠皮质神经细胞CYP2E1和iNOS的表达,降低NSE的表达;3者表达的时间变化特点基本一致,3者联合检测可用于推断乙醇的作用时间.

Expressions of CYP2E1, iNOS and NSE in primary cultured rat cortical neurons treated with ethanol

Aim:To study the changes of CYP2E1,inducible nitric oxide synthase(iNOS) and neuron-specific enolase(NSE) expressions in primary cultured rat cortical neurons treated with ethanol.Methods:Primary cultured rat cortical neurons were treated with 0,0.5,1.5 and 3.0 g/L ethanol for 30 min and 1,2,4,8,12,24 h,respectively,then the expressions of CYP2E1,iNOS and NSE in neurons were detected by immunocystochemical method.Results:Ethanol could increase the expressions of CYP2E1(Fdose=57.123,Ftime=12.111,Finteraction=4.987,P<0.001)and iNOS(Fdose=55.081,Ftime=10.949,Finteraction=4.278,P<0.001),and decrease the expression of NSE(Fdose=69.334,Ftime=10.871,Finteraction=3.252,P<0.001)in dose-dependent and time-sequential manner.The action of 1.5 g/L ethanol began at 2 h,and reached the peak at 12 h.While the action of 3.0 g/L ethanol began at 1 h,and reached the peak at 8 h.Conclusion:Ethanol could induce the expressions of CYP2E1 and iNOS,meanwhile decrease the expression of NSE in rat cortical neurons.The similar temporal patterns of CYP2E1,iNOS and NSE expressions suggest that combined detection of CYP2E1,iNOS and NSE be helpful for the action time assessment of ethanol in forensic analysis.