【摘要】 目的探讨原花青素B2（PC-B2）与黄曲霉毒素B₁（AFB₁）对人胚胎肝细胞L-02细胞色素P450（CYP）亚酶1A2、3A4活力及其基因表达的影响。方法 L-02细胞体外培养后分空白对照、溶剂对照、AFB₁染毒、PC-B2处理和PC-B2干预共5组,其中PC-B2处理分为3个亚组,分别采用3、10、30μg/ml PC-B2干预,PC-B2干预分为3各亚组,分别采用3、10、30μg/ml PC-B2预处理12 h后,再加不同浓度PC-B2与AFB₁干预24 h。测定各组细胞的CYP1A2、CYP3A4酶活力以及CYP1A2、CYP3A4基因的mRNA表达水平;测定细胞存活率,倒置荧光显微镜观察细胞形态。结果与空白对照组相比,AFB₁染毒组细胞存活率降低（P<0.05）,胞膜结构不清,漂浮细胞增多,CYP1A2、CYP3A4酶活力及其mRNA表达水平明显升高（P<0.05）。与空白对照组相比,3、10μg/ml PC-B2处理组细胞相关指标无明显变化,但30μg/ml PC-B2处理组的细胞存活率升高,CYP1A2、CYP3A4酶活力下降（P<0.05）。与AFB₁染毒比较,30μg/ml PC-B2干预组的细胞存活率升高（P<0.05）,各浓度PC-B2干预组的细胞CYP1A2、CYP3A4酶活力及基因表达均降低（P<0.05）,且干预浓度越高,CYP1A2、CYP3A4酶活力及基因表达越低（P<0.05）。结论 AFB₁能明显诱导肝细胞CYP1A2、CYP3A4酶活力及其mRNA表达,促使肝细胞凋亡,PC-B2干预可促进肝细胞生长并抑制上述CYP二个亚酶的活力及其基因表达,提示PC-B2可能通过抑制I相代谢酶CYP对AFB₁的活化而对肝细胞有良好保护作用。更多还原

【Abstract】 Objective To investigate the effects of procyanidin B2（ PC-B2） and aflatoxin B₁（ AFB₁） on the activity and gene expressions of the cytochrome P450（ CYP） 1A2 and CYP3A4 in human embryonic stem cells（ L-02）. Methods After culture in-vitro,L-02 cells were divided into 5 groups including blank control group,solvent control group,AFB₁ exposure group,PC-B2 exposure group and PC-B2 intervention group. PC-B2 exposure group was divided into 3 subgroups,and these 3 subgroups were treated with PC-B2 with the concentrations of 3,10 and30 μg / ml separately for 12 hours. PC-B2 intervention group was divided into 3 subgroups,and these 3 subgroups were treated with PC-B2 with the concentrations of 3,10 and 30 μg / ml separately for 12 hours,then were treated with PC-B2 with different concentrations and AFB₁ for 24 hours.The enzyme activities and mRNA expressions of CYP1A2 and CYP3 A were detected in each group. And the survival rate of the cells was also measured. The morphological changes of the cells were observed by fluorescent microscope. Results Compared to blank control group,the cell survival rate decreased（ P < 0. 05）,the structure of cytomembrane was unclear and floating cells increased,CYP1A2 and CYP3A4 enzyme activities and mRNA expression levels increased significantly in AFB₁ exposure group（ P < 0. 05）. No significant changes in the indices of3 μg / ml and 10 μg / ml PC-B2 exposure groups were observed compared to blank control group. But the cell survival rate increased,CYP1A2 and CYP3A4 enzyme activities decreased significantly in 30 μg / ml PC-B2 exposure group compared to blank control group（ P < 0. 05）. Compared to AFB₁ exposure group,the cell survival rate of 30 μg / ml PC-B2 intervention group increased（ P < 0. 05）,CYP1A2 and CYP3A4 enzyme activities and mRNA expressions decreased significantly in all PC-B2 intervention groups（ P < 0. 05）. And CYP1A2 and CYP3A4 enzyme activities and mRNA expressions decreased with the increase of concentration for intervention（ P < 0. 05）.Conclusion AFB₁ can induce the enzyme activities and mRNA expressions of CYP1A2 and CYP3A4 enzyme,and can promote the apoptosis of hepatic cells. Intervention with PC-B2 can promote the growth of liver cells,and can inhibit the activity and gene expression of the two CYP subenzymes. The results indicate that the protective effect of PC-B2 on hepatic cells might be mediated by inhibiting the activation of AFB₁ by CYP.更多还原