右归丸对激素型肾阳虚证miRNA表达谱的影响及其靶基因功能分析  

Effect of Yougui Pill(右归丸)on miRNA Expression Profile and Function Analysis of Target Genes in Hormone-Type Kidney Yang Deficiency Syndrome

作　　者：鲁莎 谭从娥[1] 肖雪 王相东 邢文文[1] LU Sha;TAN Cong'e;XIAO Xue;WANG Xiangdong;XING Wenwen(School of Basic Medical Sciences,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi,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：《辽宁中医杂志》2024年第5期184-187,223-224,共6页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81973754);陕西中医药大学创新团队项目(2019-QN06);咸阳市科技计划项目(2018k02-99)

摘　　要：目的 探讨微小RNA(miRNA)在右归丸对激素型肾阳虚证肾组织与正常肾组织中的表达差异,并用生物信息学方法分析其意义,为右归丸干预肾阳虚证的进一步机制研究奠定基础。方法 15只SD大鼠随机分为正常组(5只)和造模组(10只),正常组注射2 mL生理盐水,造模组后肢肌注氢化可的松注射液。成模后将造模组随机分为模型组和右归丸组,每组5只。右归丸组给予右归丸汤剂灌胃,模型组和正常组给予生理盐水灌胃。麻醉处死后使用TRIzol法提取肾组织中RNA,使用NanoDrop ND-1000对总RNA进行质量检测,用标记后的RNA与Agilent Rat 8×60k miRNA芯片进行杂交。收集原始数据进行标准化处理,进行聚类分析和火山图分析。利用生物信息学方法,预测差异miRNA富集的基因本体论(GO)功能和京都基因与基因组百科全书(KEGG)通路。结果 筛选出右归丸干预肾阳虚证相关差异表达miRNA 18条(|log_(2)FC|≥1.5,P≤0.05),其中包括5条上调miRNA,分别是rno-miR-149-3p、rno-miR-188-5p、rno-miR-221-5p、rno-miR-762、rno-miR-877;13条下调miRNA,分别是rno-miR-101b-5p、rno-miR-125a-3p、rno-miR-125b-2-3p、rno-miR-3085、rno-miR-344a-5p、rno-miR-347、rno-miR-34b-3p、rno-miR-351-3p、rno-miR-3573-3、rno-miR-370-3p、rno-miR-409a-3p、rno-miR-448-3p、rno-miR-503-3p。GO分析显示差异表达miRNA主要参与DNA、RNA转录的正调控进程且与ATP的结合显著相关。KEGG分析显示差异表达miRNA显著富集于cAMP通路和FoxO信号通路,这两条通路与线粒体功能密切相关。结论 右归丸通过干预miRNA的表达,改善肾阳虚证大鼠线粒体功能,调节能量代谢异常,进一步为右归丸干预肾阳虚证的机制研究提供理论依据。Objective To investigate the expression difference of microrNA(miRNA)in Yougui Pill(右归丸)in hormone-type kidney Yang deficiency syndrome and normal kidney tissue,and to analyze its significance by bioinformatics method,so as to lay a foundation for further study on the mechanism of Yougui Pill in the intervention of kidney Yang deficiency syndrome.Methods Fifteen SD rats were randomly divided into normal group(n=5)and model group(n=10).The normal group was injected with 2 mL normal saline,and the model group was injected with hydrocortisone injection intramuscularly.After modeling,the modeling group was randomly divided into model group and Yougui Pill group,with 5 mice in each group.Yougui Pill group was given Yougui Pill and model group and normal group were given normal saline by intragastric administration.TRIzol method was used to extract RNA from renal tissues after anesthesia.NanoDrop ND-1000 was used to detect the quality of total RNA,and the labeled RNA was hybridized with Agilent Rat 8×60k miRNA chip.The original data was collected for standardized processing,cluster analysis and volcanic map analysis.Bioinformatics methods were used to predict the gene ontology(GO)function and the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway for differential miRNA enrichment.Results Eighteen mirnas(|log_(2)FC|≥1.5,P≤0.05)related to differential expression of Yougui Pill were screened,including five up-regulated miRNAs,including rno-miR-149-3p,rno-miR-188-5p,rno-miR-221-5p,rno-miR-762 and rno-miR-877,respectively;13 down-regulated miRNAs,including rno-miR-101b-5p,rno-miR-125a-3p,rno-miR-125b-2-3p,rno-miR-3085,rno-miR-344a-5p,rno-miR-347,rno-miR-34b-3p,rno-miR-351-3p,rno-miR-3573-3,rno-miR-370-3p,rno-miR-409a-3p,rno-miR-448-3p and rno-miR-503-3p.GO analysis showed that the differentially expressed miRNAs were mainly involved in the positive regulation of DNA and RNA transcription and were significantly related to THE binding of ATP.KEGG analysis showed that differentially expressed miRNAs were significa

关 键 词：右归丸 肾阳虚证 MIRNA 能量代谢 

分 类 号：R256.5[医药卫生—中医内科学]