日本血吸虫可溶性虫卵抗原诱导肝纤维化相关miRNA的变化  

Changes of liver fibrosis-related mi RNAs induced by soluble egg antigen of Schistosoma japonicum

作　　者：王欢[1] 卢雅静[1] 高彦茹[1] 王舒弘 周蕊[1] 董惠芬[1] 

机构地区：[1]武汉大学基础医学院人体寄生虫学教研室,武汉430071

出　　处：《中国血吸虫病防治杂志》2017年第2期192-196,共5页Chinese Journal of Schistosomiasis Control

基　　金：湖北省卫生和计划生育委员会血防专项(WJ2017X001)

摘　　要：目的研究与肝纤维化相关miRNA(miR)在日本血吸虫可溶性虫卵抗原刺激小鼠肝细胞(AML12)后的表达情况,为阐明血吸虫感染导致肝纤维化的机制奠定基础。方法使用日本血吸虫可溶性虫卵抗原(Soluble egg antigens,SEA)刺激AML12后,采用定量PCR检测AML12中的miR-122、miR-182、miR-23b、miR-27b及KH型剪切调控蛋白(KHtype splicing regulatory protein,KSRP)的m NA水平,以Western blotting检测KSRP蛋白的表达变化;分别用anti-miR-27b、miR-27b precursor转染AML12后,以定量PCR和Western blotting检测AML12中KSRP的m NA和蛋白水平。结果经SEA刺激后,AML12中miR-182、miR-23b及miR-27b m RNA水平下降(P均<0.05),而细胞中miR-122 m RNA与KSRP的m RNA水平及蛋白表达水平均上调(P均<0.05)。此外,anti-miR-27b与miR-27b precursor转染组KSRP的m RNA水平均无明显变化,但anti-miR-27b组细胞中KSRP的蛋白表达增加,miR-27b precursor组KSRP的蛋白表达降低。结论SEA刺激AML12导致诸多与肝纤维化相关的miRNA及KSRP的表达发生变化,其中miR-27b可调控KSRP的表达,这为进一步研究血吸虫感染导致的肝纤维化的分子机制奠定了基础。Objective To investigate the expression of miRNA associated with hepatic fibrosis induced by Schistosoma ja-ponicum soluble egg antigen stimulation in mouse hepatocytes（AML12）,so as to lay the foundation for clarifying the mecha-nism of schistosome infection leading to hepatic fibrosis. Methods The expressions of miR-122,miR-182,miR-23b,miR-27b and KSRP in AML12 cells treated with SEA were measured by q-PCR. KSRP protein in cell lyses was measured by Western blotting. AML12 cells were transfected with miR-27b precursor or anti-miR-27b for 24 h,then q-PCR was adopted to determine KSRP mRNA,and KSRP protein was detected by Western blotting. Results The expressions of miR-182,miR-23b and miR-27b were decreased and miR-122 was increased in AML12 cells following SEA treatment（all P〈0.05）. An increase of mRNA and protein of KSRP expression was also observed in AML12 cells after SEA stimulation（both P〈0.05）. In addition,KSRP mRNA expression was not changed significantly in AML12 cells transfected with anti-miR-27b or miR-27b precursor,and miR-27b precursor reduced KSRP protein expression as compared with the control. In contrast,the expression of KSRP protein was increased in the anti-miR-27b group and decreased in the miR-27b precursor group. Conclusions After the stimulation of SEA,the expressions of a variety of liver fibrosis-related miRNAs and KSRP change in murine hepatocytes,including miR-27b. And miR-27b can regulate the expression of KSRP. These findings might lay a foundation for further study on the molecular mechanism of fibrosis induced by schistosome infection.

关 键 词：日本血吸虫 肝纤维化 MIRNA 可溶性虫卵抗原 肝细胞 

分 类 号：R383.24[医药卫生—医学寄生虫学]