piRNA在小鼠非酒精性脂肪性肝病中的表达谱分析  

Analysis of piRNA expression spectra in a nonalcoholic fatty liver disease mouse model

作　　者：马旭阳 李昌平 MA Xu-yang;LI Chang-ping(Department of Gastroenterology,the Affiliated Hospital of Southwest Medical University,Luzhou,Sichuan,646000,China)

机构地区：[1]西南医科大学附属医院消化内科,646000

出　　处：《现代消化及介入诊疗》2020年第4期459-466,471,共9页Modern Interventional Diagnosis and Treatment in Gastroenterology

摘　　要：目的研究piRNA在非酒精性脂肪性肝病(NAFLD)中的表达情况及两者之间的潜在关系。方法将雄性C57BL/6小鼠随机分为对照组和模型组,每组10只。对照组进食蛋氨酸胆碱充足饲料,模型组进食蛋氨酸胆碱缺乏饲料。采用苏木精-伊红染色对肝组织切片行病理评估,并检测血清肝功能指标,包括谷丙转氨酶(ALT)、谷草转氨酶(AST)、甘油三酯(TG)和胆固醇(TC)。使用piRNA基因芯片初筛肝组织中的差异piRNA,然后对差异piRNA行生物信息学分析,通过逆转录-定量聚合酶链反应进一步验证piRNA的表达。结果模型组小鼠建立了有效的NAFLD模型。与对照组相比,模型组小鼠血清AST、ALT明显增高,TC、TG降低。模型组中共有1285个差异表达的piRNA,其中有641个piRNA发生上调,644个piRNA发生下调。生物信息学分析表明,差异表达的piRNA主要富集于hippo、Wnt、Cushing syndrome、FoxO、MAPK等信号通路。选择部分piRNA行RT-qPCR分析,其中piRNA-DQ566704、piRNA-DQ723301的结果与基因芯片分析结果基本一致。结论在NAFLD发生发展过程中,确实存在piRNA的差异表达,然而piRNA影响NAFLD发展的具体机制还有待进一步研究。Objective Nonalcoholic fatty liver disease(NAFLD)has become a common health issue worldwide,and piRNA has been shown to be differentially expressed in a variety of diseases.The purpose of this research was to explore the potential relationship between P-element-induced wimpy testis interacting RNA(piRNA)and NAFLD.Methods The NAFLD model of mice was established by methionine-and choline-deficient(MCD)diet and methionine-and choline-sufficient(MCS)diet.Following this step,the mouse liver tissues were removed to be stained with haematoxylin eosin(HE),and the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC)and triglyceride(TG)expression were measured.The liver tissues of the control group and model group were selected for piRNA gene chip analysis to find piRNAs with differential expression in NAFLD.The differentially expressed piRNAs screened from the microarray were verified by reverse transcription-quantitative polymerase chain reaction(RT-qPCR).piRNAs with potential research value were selected for further analysis of target genes,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways.Results In this study,we found a total of 1285 piRNAs with different expression levels.The results showed that in the model group,641 piRNAs were highly expressed,while 644 piRNAs were underexpressed.The piRNAs were enriched in the cancer,Hippo signalling,Wnt signalling,and MAPK signalling pathways.The RTq PCR results showed that piRNA DQ566704 and piRNA DQ723301 were significantly upregulated in the model group.Conclusion Our study confirms that differentially expressed piRNAs exist in NAFLD and may affect the development of NAFLD through related pathways.

关 键 词：非酒精性脂肪性肝病 非酒精性脂肪性肝炎 PIRNA 蛋氨酸胆碱缺乏 蛋氨酸胆碱充足 piRNA基因芯片 

分 类 号：R575.5[医药卫生—消化系统]