中文摘要：

目的探讨caspase-12表达变化与内质网应激介导小鼠原代肝细胞凋亡的关系。方法原位消化灌流法分离小鼠原代肝细胞，应用毒胡萝卜索对小鼠原代肝细胞进行凋亡诱导，通过DNA梯带，Hoechst染色和流式细胞仪等对肝细胞凋亡进行定性和定量检测，选出合适的诱导浓度和诱导时间。用Western印迹法检测凋亡过程中caspase-12蛋白酶的表达变化。结果毒胡萝卜素4μmol、诱导30h可引起50％肝细胞发生凋亡，并出现典型的凋亡形态改变。随诱导时间延长procaspase-12蛋白表达逐渐减少。结论毒胡萝卜素4μmol作用30h是建立小鼠原代肝细胞凋亡模型的合适条件，随凋亡细胞数量增加，procaspase-12相应减少，caspase-12活化是毒胡萝卜素引起内质网应激介导肝细胞凋亡的主要机制。

英文摘要：

Objective To investigate the relationship between caspase-12 activation and ER stress mediated apoptosis of primary mouse hepatocytes. Methods Mouse hepatocytes were isolated by modified 2-step collagenase perfusion method. The optimal TG concentration and incubation time for inducing cell apoptosis was determined by morphological analysis and agarose gel electrophoresis presented as cell apoptotie percentage in DNA ladder and flow cytometry. Expression of procaspase-12 during the process was measured by Western Blot. Results When the primary mouse hepatocytes were treated with 4 μmol TG, the percentage of apoptotic cells increased. The percentage of apoptosis was 50% at the concentration of 4 μmol of TG after 30 hours incubation. The protein level of proeaspase-12 determined by Western Blot declined in a time dependent manner. Conclusion 4μmol TG incubation for 30 hours can induce half of the primary hepatocytes underwent apoptosis. Proeaspase-12 was activated into a cleaved form resulting in decrease of procaspase-12. These results suggest thai procaspase-12 activation is closely related with ER stress mediated apoptosis and might play an important role in that.