胆汁淤积性肝损伤大鼠肝脏组织中水通道蛋白8/9表达的变化  

作　　者：叶蕾[1] 张婷婷 曹婕露 陈曦[1] 胡侃男 姚嘉明[2] 陈芝芸[1] 严峻彬 

机构地区：[1]浙江中医药大学附属第一医院,310006 [2]杭州市中医院,310007

出　　处：《浙江临床医学》2021年第8期1138-1141,共4页Zhejiang Clinical Medical Journal

基　　金：浙江省中医药科技计划资助项目(2018ZA040);浙江省科技计划项目(2014C33238)。

摘　　要：目的探讨α-异硫氟酸蔡酯(ANIT)诱导的胆汁淤积性肝损伤大鼠肝组织中水通道蛋白8(AQP8)与水通道蛋白9(AQP9)的表达及其作用机制。方法48只SD大鼠随机分为模型组和对照组,每组各24只。模型组大鼠以麻油配制的4%ANIT一次性灌胃,对照组以等量麻油灌胃,分别于灌胃后24 h、48 h、72 h处理大鼠,每个亚组各8只。采用全自动生化分析仪检测大鼠的血清ALT、AST、ALP、TBIL、TEA水平,免疫组化检测肝组织CK19表达,荧光定量PCR检测肝组织PI3K、ART、AQP8和AQP9mRNA表达,Western blotting检测肝组织PI3K、AKT、AQP8、AQP9蛋白表达和AKT蛋白磷酸化水平。结果模型组大鼠灌服ANTI后,其血清ALT、AST、ALP、TBIL、TEA水平均较对照组显著升高,PI3K、AKT、AQP8、AQP9的mRNA和蛋白表达及AKT磷酸化水平下调;上述变化均于48 h最为明显,72 h有所改善。结论ANTI诱导的胆汁淤积性肝损伤大鼠模型会出现PI3K/AKT信号通路的抑制,以及AQP8、AQP9表达的下调。Objective To investigate the α-naphthyl isothiocyanate(ANIT)-induced liver tissue aquaporin 8(AQP8)and aquaporin 9(AQP9)expression and its mechanism of cholestatic liver injury in rats.Methods Forty-eight SD rats were randomly divided into model group and control group,with 24 rats in each group.The rats in the model group were given 4%ANIT prepared with sesame oil once by gavage,and the rats in the control group were given the same amount of sesame oil by gavage.The rats were treated 24 h,48 h,and 72 h after gavage,8 rats in each subgroup.CK19 expression in hepatic tissue was detected by Immunohistochemistry,ALT、AST、ALP、TBIL、TBA were observed by automatic biochemistry analyzer,PI3K、AKT、AQP8 and AQP9 mRNA expression in hepatic tissue were analyzed by quantitative fluorogenic PCR,PI3K、AKT、AQP8、AQP9 protein expression in hepatic tissue and the phosphorylation level of AKT were detected by Western blotting.Results After gavage ANTI,the model group showed significant increases in the serum levels of ALT、AST、ALP、TBIL,and TBA compared with the healthy control group.The model group showed down-regulation of PI3K、AKT、AQP8、AQP9mRNA and protein expression,decreasing the phosphorylation level of AKT.The above changes were the most serious at 48 hours,recovered at 72 hours.Conclusion The inhibition of PI3K/AKT signaling pathway and the downregulation expression of AQP8 and AQP9 can be observed in ANTI induced cholestatic liver injury rats.

关 键 词：胆汁淤积 肝损伤 PI3K/AKT信号通路 水通道道白8 水通道蛋白9 

分 类 号：R73[医药卫生—肿瘤]