大鼠失血性休克及复苏后肝脏EPHX2和Sp1基因表达的变化

Expression of EPHX2 and Sp1 gene in rat liver following severe hemorrhagic shock and resuscitation

目的观察失血性休克及复苏对大鼠肝脏EPHX2基因及预测转录因子Sp1表达的影响。方法通过软件预测EPHX2转录因子,筛选出Sp1为研究对象。将25只SD大鼠随机平均分为5组:对照(CON)组、失血性休克(HS)组、休克复苏1 h(HSR1)组、休克复苏3 h(HSR3)组、休克复苏6 h(HSR6)组。采用real-time RT-PCR和蛋白质印迹法检测大鼠肝脏中EPHX2和Sp1的表达。结果HS、HSR1、HSR3、HSR6组EPHX2 mRNA表达低于对CON组(P<0.05),HSR1、HSR3、HSR6组EPHX2 mRNA表达低于HS组(P<0.05),HSR1组EPHX2 mRNA表达低于HSR3、HSR6组(P<0.05)。CON组Sp1mRNA表达和其他组比较差异无统计学意义(P>0.05),HS、HSR1组Sp1 mRNA表达低于HSR6组(P<0.05)。HSR1、HSR3、HSR6组EPHX2蛋白表达较CON组降低(P<0.05),Sp1蛋白在各组间表达差异无统计学意义。结论失血性休克可引起肝脏EPHX2基因表达下调,而复苏不能快速逆转这种趋势;失血性休克及复苏对预测转录因子Sp1表达没有影响。

Objective To evaluate the effect of severe hemorrhagic shock and resuscitation on expression of EPHX2 and predictive transcription factor Sp1 in rat liver.Methods Sp1 was selected for study by software prediction of the transcription factor EPHX2.A total of 25 SD rats were randomly divided into 5 groups（n = 5） ： control（CON） group,hemorrhagic shock（HS） group,1 h after hemorrhagic shock and resuscitation（HSR1） group,3 h after hemorrhagic shock and resuscitation（HSR3） group,and 6 h after hemorrhagic shock and resuscitation（HSR6） group.Real-time RT-PCR and Western blotting analysis were used to examine the expression of EPHX2 and Sp1 in the rat liver.Results The expressions of EPHX2 mRNA in HS,HSR1,HSR3,and HSR6 groups were significantly lower than those in CON group（P〈0.05）,those in HSR1,HSR3 and HSR6 groups were significantly lower than those in HS group（P〈0.05）,and that in HSR1 group was significantly lower than those in the HSR3 and HSR6 groups（P〈0.05）.The expression of Sp1 mRNA in CON group was not significantly different from those in other groups,but the expressions in HS and HSR1 groups were significantly lower than that in the HSR6 group（P〈0.05）.Compared with CON group,EPHX2 protein expression was significantly lower in HSR1,HSR3,and HSR6 groups（P〈0.05）,and Sp1 protein expressions were similar in different groups.Conclusion EPHX2 mRNA expression is decreased in rat liver following severe hemorrhagic shock,and this decrease cannot be reversed by effective resuscitation;Sp1 is not involved in the process.