肾阳虚证差异表达miRNA筛选及其作用通路分析  

Screening and Pathway Analysis of Differentially Expressed miRNA in Kidney Yang Deficiency Syndrome

作　　者：肖雪 鲁莎 谭从娥[1] 邢文文[1] 王相东 XIAO Xue;LU Sha;TAN Cong-e;XING Wen-wen;WANG Xiang-dong(Basic Medical College of Shaanxi University of Chinese Medicine yXianyang,Shaanxi 712046,China)

机构地区：[1]陕西中医药大学基础医学院,陕西咸阳712046

出　　处：《时珍国医国药》2022年第4期1011-1014,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81973754);陕西中医药大学创新团队项目(2019-QN06)。

摘　　要：目的筛选肾阳虚证相关差异表达miRNA及其靶基因,探讨靶基因生物学功能及其作用的分子通路。方法将10只雄性SD大鼠随机分为正常组和模型组,每组5只,模型组大鼠每日一次给予氢化可的松注射2.5mg/100g后肢肌肉注射,正常组大鼠每日一次给予0.9%生理盐水2ml后肢肌肉注射,连续注射2周。造模成功后采集两组大鼠的肾皮质,提取组织RNA,采用Agilent miRNA芯片检测两组肾皮质中的差异表达miRNA。使用miWALK数据库预测差异表达miRNA的靶基因,利用DAVID Bionformatica Resources 6.8和Metascape数据库分析其靶基因的生物学功能和信号通路。结果筛选出36条显著差异表达miRNAs,其中23条miRNAs表达上调,13条miRNAs表达下调。这些差异表达miRNA靶基因与细胞质、轴突、高尔基体、囊泡介导运输、细胞内蛋白质运输、蛋白激酶结合等生物学功能相关,其富集的信号通路主要有自噬通路、胞吞作用、癌症通路、癌症中的蛋白聚糖、环腺苷酸信号通路、HIF-1信号通路,其中自噬通路最具显著性。结论自噬功能相关miRNA及其介导的信号通路可能与肾阳虚证的发生密切相关。Objective To screen differentially expressed miRNAs and their target genes related to kidney Yang deficiency syndrome,and to explore the biological function and molecular pathways of target genes.Methods Ten male SD rats were randomly divided into the normal group and the model group,with five rats in each group.The rats in the model group were injected intramuscularly with hydrocortisone at 2.5 mg/100 g once a day,and the rats in the normal group were injected intramuscularly with 0.9%normal saline at 2 mL once a day for two weeks.After successful modeling,the renal cortex of the two groups of rats was collected,and the tissue RNA was extracted.Agilent miRNA chip was used to detect the differential expression of miRNA in the renal cortex of the two groups.MiWALK database was used to predict the target genes of differentially expressed miRNAs.DAVID Bioniformatical Resources 6.8 and Metascape database were used to analyze the biological functions and signaling pathways of the target genes.Results 36 differentially expressed miRNAs were screened,of which 23 were up-regulated and 13 were down-regulated.These differentially expressed miRNA target genes are related to biological functions such as cytoplasm,axon,Golgi apparatus,vesicle-mediated transport,intracellular protein transport,and protein kinase binding.The enriched signaling pathways mainly include autophagy pathway,endocytosis,pathway in cancer,proteoglycans in cancer,CAMP signaling pathway,and HIF-1 signaling pathway,among which autophagy pathway is the most significant.Conclusion Autophagy related miRNA and its mediated signaling pathway may be closely related to the occurrence of kidney Yang deficiency syndrome.

关 键 词：肾阳虚证 MIRNA 自噬 衰老 

分 类 号：R285.5[医药卫生—中药学]