黄芪甲苷对缺血性急性肾损伤大鼠模型长链非编码RNA表达谱的影响

Screening and functional studies of long non - coding RNA in protecting ischemic acute kidney injury by Astragaloside Ⅳ

目的研究采用长链非编码RNAs(LncRNAs)测序技术筛选黄芪甲苷(AS-IV)干预急性肾损伤(AKI)大鼠过程中肾组织相关LncRNAs的变化.方法选取雄性SD大鼠8只,采用随机数字表法分成两组,AKI模型组(组1):4只大鼠进行缺血再灌注(I/R)处理;AS-IV预处理组(组2):4只大鼠I/R处理前7 d分别进行AS-IV灌胃处理,I/R处理24 h后取肾组织,提取总RNA并进行质量检测,合格后进行测序实验,以差异倍数(组1/组2)≥2且P≤0.05为纳入标准,检测AKI模型组和AS-IV预处理组中的LncRNAs差异表达谱,并对差异LncRNAs进行基因本体论(GO)分析和通路(Pathway)分析,从而了解LncRNAs参与的生物学功能.结果两组相比,差异表达的LncRNAs共232条,其中127条表达上调,105条表达下调.发现341条mRNA表达出现差异性变化,其中178条表达上升和163条表达下降.通过GO分析获得mRNA参与的细胞生物过程、细胞组件和分子功能,LncRNAs共表达的mRNA主要参与核小体组装、染色质组装或分解、核小体组织、DNA构象变化、蛋白质-DNA复合物组装等生物过程,参与的细胞组件包括核小体、蛋白质-DNA复合物、核染色质、染色质和核核小体等成分,体现了蛋白质异源二聚反应活性、蛋白二聚化活性、DNA结合活性和核酸结合活性等分子功能.细胞通路分析LncRNAs共表达的mRNAs所参与的信号通路主要涉及到系统性红斑狼疮途径、酒精代谢途径、病毒致癌机制等,共同调控着黄芪甲苷保护AKI的生物学进程.结论 AKI模型组和AS-IV预处理组大鼠肾组织中存在差异表达的LncRNAs,生物学功能和通路分析提示这些差异表达LncRNAs可能参与黄芪甲苷改善AKI的作用机制.

Objective To investigate the expression profile of long non - coding RNAs(lncRNAs) in acute kidney injury (AKI) rats upon astragaloside IV(AS - IV) treatment. Methods Eight male SD rats were selected and randomly divided into two groups. AKI model group(group 1):4 rats were subjected to ischemia - reperfusion(I/ R);AS -Ⅳ pretreatment group (group 2):4 rats were orally administered AS -Ⅳ for 7 days prior to I/ R. Renal tissues were collected after I/ R treatment of 24h,total RNA was extracted from renal tissues and tested for quality. Sequencing experiments were carried out after being qualified. The threshold set for up - and down - regulated genes was a fold change(group1 / group2)≥2 and P≤0. 05. Afterwards,GO analysis and KEGG analysis were used to determine the roles that these differentially expressed mRNAs played in these GO terms or pathways. Results Two hundred and thirty - two lncRNAs were differentially expressed(127 lncRNAs were up - regulated and 105 lncRNAs were down -regulated). Three hundred and forty - one mRNAs were differentially expressed(178 mRNAs were up - regulated and 163 mRNAs were down - regulated). GO analysis indicated that differentially expressed mRNAs were mainly involved in biological processes such as nucleosome assembly,chromatin assembly,nucleosome organization,chromatin assembly or disassembly and DNA conformation change. GO analysis indicated that differentially expressed mRNAs were mainly involved in cellular components such as nucleosome,protein - DNA complex,nuclear chromatin,chromatin and nuclear nucleosome. GO analysis indicated that differentially expressed mRNAs were mainly involved in molecular functions such as protein heterodimerization activity,protein dimerization activity,DNA binding,nucleic acid binding. Signal pathway analysis indicated that lncRNAs and mRNAs were involved in systemic lupus erythematosus,alcoholism and viral carcinogenesis. Conclusion LncRNAs expression differed significantly in renal tissues of AKI model group and AS -Ⅳ preconditioning group. Study of the biological functions and pathways of these lncRNAs indicated that they may be involved in the mechanism of AS -Ⅳ ameliorated ischemic acute renal injury.