文献类型：期刊文献

中文题名：心肌成纤维细胞来源的外泌体miRNA参与放射性心肌纤维化损伤

英文题名：Exosomes MicroRNA Derived from Cardiac Fibroblasts Involved in Radioactive Cardiac Fibrosis Injury

作者：段依璠[1,2];顾静[1,3];舒亚妃[4];韩晓斐[4,5];梁乾坤[3,5]

第一作者：段依璠

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]天津医科大学第二医院血液内科,兰州730000;[3]甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室,兰州730000;[4]甘肃省中医方药挖掘与创新转化重点实验室,兰州730000;[5]甘肃省中医药防治慢性疾病重点实验室,兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2023

卷号：39

期号：1

起止页码：108

中文期刊名：中国生物化学与分子生物学报

外文期刊名：Chinese Journal of Biochemistry and Molecular Biology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD_E2023_2024】；PubMed;

基金：甘肃省“双一流”科研重点项目(No.GSSYLXM-05);甘肃省中医药管理局重点项目“甘肃省中医药科研课题”(No.GZKZ-2020-10);甘肃省教育科技创新项目“青年博士基金”(No.2022QB-094);甘肃省自然科学基金(No.21JR1RA262);甘肃省中医药研究中心“中医方药挖掘与创新转化重点实验室”开放课题(No.ZYFY-2020-006)。

语种：中文

中文关键词：放射性心肌纤维化;心肌成纤维细胞;外泌体;微RNA;生物信息学分析

外文关键词：radiation-induced cardiac fibrosis(RICF);cardiac fibroblasts(CF);exosomes;microRNA(miRNA);bioinformatics analysis

摘要：外泌体及携带的miRNA在组织器官纤维化中发挥重要作用,但相关于其在放射性心肌纤维化(RICF)的研究十分有限,本研究拟运用生物信息学分析辐射诱导的外泌体miRNA通过辐射旁效应促进RICF发生的可能分子机制。心肌成纤维细胞(CFs)是RICF的主要效应细胞,用2 Gy X射线辐照CFs后超速离心提取辐射诱导的外泌体(X-exo)和未受照射的CFs外泌体(Exo),并进行外泌体电镜观察、NTA浓度检测和CD9、CD63、CD81等外泌体表面标志蛋白质的纳米流式鉴定;随后通过RNA-seq技术检测外泌体miRNA表达谱,筛选差异表达的miRNA,预测其潜在靶基因并进行富集分析。结果显示:与未受照射的CFs外泌体相比,X射线诱导的外泌体中表达上调的miRNAs共9个(|log_(2) Fold Change|>1,P<0.05),其中|log_(2) Fold Change|>2的有8个,表达下调的miRNAs共19个(|log_(2) Fold Change|>1,P<0.05),其中|log_(2) Fold Change|>2的有12个。使用targetScan、miRWalk、miRNADB预测差异miRNA的靶基因,并进行GO及KEGG富集分析。GO富集结果显示,差异表达的miRNA调控的靶基因主要参与蛋白质磷酸化、细胞信号转导等生物学过程,富集于细胞质和细胞膜等细胞组分,发挥蛋白激酶绑定和蛋白质结合等分子功能。KEGG富集结果显示,差异表达miRNAs的靶基因主要富集于PI3K-Akt、MAPK、mTOR、ECM-receptor interaction、cAMP、Wnt、TGF-β、Notch等相关通路。对本研究富集到的信号通路进行文献梳理,表明差异表达的外泌体miRNA可能通过调节靶基因及相关信号通路促进RICF的发生发展。因MAPK、PI3k-Akt、mTOR等信号通路的活化调节主要依靠关键分子的磷酸化,并非主要依靠转录水平的调控,所以未来在探讨“外泌体miRNA通过辐射旁效应参与RICF”的机制研究中,应该着重关注ECM、cAMP、TGF-β、Wnt、Notch、Ras、Rap1等信号通路。
In recent years,it has been found that microRNA(miRNA) carried by exosomes play an important role in the development of tissue and organ fibrosis.However,studies on the relationship between radiation-induced cardiac fibrosis(RICF) and exosomes and their miRNA are very limited.The purpose of this study is to analyze the possible role of radiation-induced exosome miRNA in the development of RICF by bioinformatics.Myocardial fibroblasts(CFs) are the main effector cells of RICF.The CFs were irradiated with 2 Gy X-rays,and radiation-induced exosomes(X-exo) and unirradiated exosomes of CFs(Exo) were extracted by overspeed centrifugation.Then,exosomes were observed and identified with its morphology,NTA concentration,and exosome surface marker proteins such as CD9,CD63,and CD81.Subsequently,RNA-seq technology was used to detect the miRNA expression profiles of Exo and X-exo,then the differentially expressed miRNAs were screened and their potential target genes and enrichment analysis were analyzed.The results showed that,compared with the control group(Exo),9 miRNAs were up-regulated in the X-exo(|log_(2 )Fold Change|>1,P2;19 miRNAs were down-regulated in the X-exo(|log_(2 )Fold Change|>1,P2.TargetScan,miRWalk and miRNADB were used to predict the target genes of differentially expressed miRNAs,and GO and KEGG enrichment analysis were performed.GO enrichment results showed that target genes regulated by differentially expressed miRNAs were mainly involved in biological processes such as protein phosphorylation and cell signal transduction.Those differentially expressed miRNAs were enriched in cytoplasm,cell membrane and other cellular components,playing molecular functions such as protein kinase binding and protein binding.KEGG enrichment results showed that target genes with differentially expressed miRNAs were mainly enriched in PI3K-Akt,MAPK,mTOR,ECM-receptor interaction,cAMP,Wnt,TGF-β and Notch related signaling pathways.Combing with literatures,it is suggested that differentially expressed exosomal miRNA may promote the development of RICF by regulating the target genes and related signaling pathways.Because the activation regulation of signaling pathways such as MAPK,PI3K/AKT,and mTOR are mainly dependent on the phosphorylation of key molecules,but not on the regulation of the transcriptional level.The detailed mechanism of "Exosomes miRNA mediate radiation bystander effects to promote RICF" will be investigated in future studies,and the signaling pathways such as ECM,cAMP,TGF-β,Wnt,Notch,Ras,and Rap1 should be the focus.