GRP78、GRP94及CHOP在亚砷酸钠致肝损伤过程中的表达变化

Change of mRNA and protein levels of GRP78,GRP 94 and CHOP in the process of sodium arsenite-induced liver injury

目的观察亚砷酸钠致肝损伤过程中肝脏内GRP78、GRP94及CHOP的表达变化并探讨其在亚砷酸钠致肝损伤中的可能作用。方法 Wistar大鼠20只,随机分为正常组及亚砷酸钠组,每组各10只。亚砷酸钠组大鼠自由饮用含200mg/L的NaAsO2自来水制备砷中毒致肝损伤模型,正常组大鼠则自由饮用自来水,时间为8周。采用免疫组织化学技术检测肝组织中GRP78、GRP94及CHOP蛋白的表达变化;采用PCR技术检测肝脏中GRP78、GRP94及CHOP mRNA的变化;同时应用TUNEL法检测肝细胞凋亡。结果免疫组织化学技术检测发现亚砷酸钠组大鼠肝脏中GRP78、GRP94及CHOP在蛋白水平的表达较正常组大鼠显著增加(P<0.05);PCR检测结果显示亚砷酸钠组大鼠肝脏中GRP78、GRP94及CHOP mRNA的表达也较正常组显著增加(P<0.05);此外与正常组比较,亚砷酸钠组大鼠肝脏中肝细胞凋亡明显增多(P<0.05)。结论 GRP78、GRP94及CHOP在基因及蛋白水平表达上调可能参与了亚砷酸钠致肝损伤的发生发展过程。

Objective To measure the change of mRNA and protein levels of GRP78,GRP 94 and CHOP in the process of sodium arsenite-induced liver injury and further to explore their possible roles in liver damage.Methods 20 Wistar rats were randomly divided into normal control group and arsenic poisoning model group,10 in each group.For the next 8weeks,the rats in model group were fed with tap water containing 200mg/L NaAsO2 for the establishment of arsenic poisoning model,while the rats in control group were fed with tap water.mRNA and protein levels of GRP78,GRP 94 and CHOP were detected by real-time PCR and immunohistochemical techniques,respectively.Meanwhile,cell apoptosis in liver was detected by TUNEL.Results Compared with normal control group,both mRNA and protein levels of GRP78,GRP 94 and CHOP in rats of arsenic poisoning model group increased significantly（P〈0.05）.Besides,the apoptosis of hepatocytes in arsenic poisoning model group was elevated（P〈0.05）.Conclusion The up-regulating of mRNA and protein levels of GRP78,GRP 94 and CHOP might play important roles in the process of sodium arsenite-induced liver injury.