發(fā)布時間：2018-02-27 11:03

  本文關(guān)鍵詞： 日本血吸蟲 肝纖維化 miRNA 可溶性蟲卵抗原 肝細(xì)胞　出處：《中國血吸蟲病防治雜志》2017年02期 　論文類型：期刊論文

【摘要】：目的研究與肝纖維化相關(guān)miRNA(miR)在日本血吸蟲可溶性蟲卵抗原刺激小鼠肝細(xì)胞(AML12)后的表達(dá)情況,為闡明血吸蟲感染導(dǎo)致肝纖維化的機(jī)制奠定基礎(chǔ)。方法使用日本血吸蟲可溶性蟲卵抗原(Soluble egg antigens,SEA)刺激AML12后,采用定量PCR檢測AML12中的miR-122、miR-182、miR-23b、miR-27b及KH型剪切調(diào)控蛋白(KHtype splicing regulatory protein,KSRP)的m NA水平,以Western blotting檢測KSRP蛋白的表達(dá)變化;分別用anti-miR-27b、miR-27b precursor轉(zhuǎn)染AML12后,以定量PCR和Western blotting檢測AML12中KSRP的m NA和蛋白水平。結(jié)果經(jīng)SEA刺激后,AML12中miR-182、miR-23b及miR-27b m RNA水平下降(P均0.05),而細(xì)胞中miR-122 m RNA與KSRP的m RNA水平及蛋白表達(dá)水平均上調(diào)(P均0.05)。此外,anti-miR-27b與miR-27b precursor轉(zhuǎn)染組KSRP的m RNA水平均無明顯變化,但anti-miR-27b組細(xì)胞中KSRP的蛋白表達(dá)增加,miR-27b precursor組KSRP的蛋白表達(dá)降低。結(jié)論SEA刺激AML12導(dǎo)致諸多與肝纖維化相關(guān)的miRNA及KSRP的表達(dá)發(fā)生變化,其中miR-27b可調(diào)控KSRP的表達(dá),這為進(jìn)一步研究血吸蟲感染導(dǎo)致的肝纖維化的分子機(jī)制奠定了基礎(chǔ)。
[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.
【作者單位】： 武漢大學(xué)基礎(chǔ)醫(yī)學(xué)院人體寄生蟲學(xué)教研室;
【基金】：湖北省衛(wèi)生和計劃生育委員會血防專項(xiàng)(WJ2017X001)
【分類號】：R532.21;R575.2
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本文編號：1542395