日本血吸虫可溶性虫卵抗原诱导肝纤维化相关miRNA的变化

发布时间：2018-02-27 11:03

  本文关键词： 日本血吸虫 肝纤维化 miRNA 可溶性虫卵抗原 肝细胞　出处：《中国血吸虫病防治杂志》2017年02期 　论文类型：期刊论文

【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。
[Abstract]:Objective to study the expression of miRNAmiR associated with hepatic fibrosis in mice liver cells stimulated by soluble egg antigen of Schistosoma japonicum (Schistosoma japonicum). Methods the AML12 was stimulated by Schistosoma japonicum soluble egg antigen Soluble egg antigens (SEAA) and the mRNA levels of miR-122 miR-182 miR-23btmiR-27b and KHHtype splicing regulatory protein KSRP in AML12 were detected by quantitative PCR method in order to elucidate the mechanism of hepatic fibrosis induced by Schistosoma japonicum infection. Western blotting was used to detect the expression of KSRP protein, and anti-miR-27btmiR-27b precursor was used to transfect AML12. Results after stimulation with SEA, the levels of miR-182 miR-23b and miR-27b m RNA in AML12 were decreased (P 0.05), while the level of m RNA and protein expression of miR-122 m RNA and KSRP were all up-regulated (P 0.05). There was no significant change in m RNA level in KSRP transfected with external anti-miR-27b and miR-27b precursor. However, the protein expression of KSRP in anti-miR-27b group increased the expression of KSRP protein in precursor group of miR-27b. Conclusion the expression of miRNA and KSRP related to liver fibrosis can be changed by SEA stimulation of AML12, in which miR-27b can regulate the expression of KSRP. This lays a foundation for further studying the molecular mechanism of hepatic fibrosis caused by schistosomiasis infection.
【作者单位】： 武汉大学基础医学院人体寄生虫学教研室;
【基金】：湖北省卫生和计划生育委员会血防专项(WJ2017X001)
【分类号】：R532.21;R575.2
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