Article(id=1203053368743781328, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1203053366290113441, articleNumber=null, orderNo=null, doi=10.11855/j.issn.0577-7402.2023.03.0275, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1644681600000, receivedDateStr=2022-02-13, revisedDate=null, revisedDateStr=null, acceptedDate=1659974400000, acceptedDateStr=2022-08-09, onlineDate=1764759874759, onlineDateStr=2025-12-03, pubDate=1679932800000, pubDateStr=2023-03-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1764759874759, onlineIssueDateStr=2025-12-03, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1764759874759, creator=13701087609, updateTime=1764759874759, updator=13701087609, issue=Issue{id=1203053366290113441, tenantId=1146029695717560320, journalId=1189873630562394117, year='2023', volume='48', issue='3', pageStart='245', pageEnd='366', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1764759874174, creator=13701087609, updateTime=1764810242575, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1203264626747220064, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1203053366290113441, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1203264626747220065, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1203053366290113441, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=275, endPage=282, ext={EN=ArticleExt(id=1203053369129657310, articleId=1203053368743781328, tenantId=1146029695717560320, journalId=1189873630562394117, language=EN, title=MiR-149-5p targeting FGF21 promotes hypoxia/reoxygenation injury of rat's cardiomyocytes, columnId=1190310110212751762, journalTitle=Medical Journal of Chinese People’s Liberation Army, columnName=Basic Research, runingTitle=null, highlight=null, articleAbstract=

Objective To investigate the targeting relationship of miR-149-5p and fibroblast growth factor 21 (FGF21), and the effect of miR-149-5p on hypoxia/reoxygenation (H/R) injury of rat`s cardiomyocytes. Methods Ten male C56BL/6J mice were used to construct the cardiac ischemia-reperfusion (I/R) models, and qRT-PCR was performed to detect the changes of miR-149-5p levels in the heart tissues of I/R mice. H9c2 cells cultured to logarithmic phase were divided into control group, H/R group, H/R+miR-149-5p inhibitor group, H/R+miR-149-5p NC group, H/R+miR-149-5p mimics group and H/R+miR-149-5p mimics+FGF21 group, and induced H/R model and transfection treatment. The activities of lactate dehydrogenase (LDH) and creatine kinase (CK) in H9c2 cardiomyocytes were detected by ELISA; Cell viability was detected by MTT assay; apoptosis was detected by flow cytometry; the level of miR-149-5p was detected by qRT-PCR; the expression levels of Bcl-2, Bax, cleaved caspase-3 and FGF21 proteins were detected by Western blotting; the targeting relationship between miR-149-5p and FGF21 were determined by dual luciferase assay. Results Compared with the sham operation group, the level of miR-149-5p in the heart tissue of I/R mice was significantly increased (P<0.05). Compared with those in control group, the apoptosis rate, LDH and CK activities and Bax, cleaved caspase-3 protein expression levels of H9c2 cells in H/R group were significantly increased (P<0.05), cell viability and FGF21, Bcl-2 protein expression levels decreased significantly (P<0.05). Compared with those in H/R group and H/R+miR-149-5p NC group, the apoptosis rate, LDH and CK activities and Bax, cleaved caspase-3 protein expression levels of H9c2 cells in H/R+miR-149-5p inhibitor group were significantly decreased (P<0.05), cell viability and FGF21, Bcl-2 protein expression levels were significantly increased (P<0.05). Compared with those in H/R group, the apoptosis rate, LDH and CK activities and Bax, cleaved caspase-3 protein expression levels of H9c2 cells in H/R+miR-149-5p mimics group were significantly increased (P<0.05), cell viability and FGF21, Bcl-2 protein expression levels decreased significantly (P<0.05). Compared with those in H/R+miR-149-5p mimics group, the apoptosis rate, and LDH and CK activities of H9c2 cells in miR-149-5p mimics+FGF21 group were significantly decreased (P<0.05), cell viability was significantly increased (P<0.05). The result of dual luciferase assay indicated the targeted regulatory relationship between miR-149-5p and FGF21. Conclusions MiR-149-5p is up-regulated significantly in the myocardial tissue of I/R mice, and negatively regulate the level of FGF21, leads to decreased cell viability and increased apoptosis of H/R-treated rat's cardiomyocytes.

, correspAuthors=Ying Zhang, authorNote=null, correspAuthorsNote=, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Xian-Xun Jiang, Kai Zhang, Ying Zhang), CN=ArticleExt(id=1203053372535431276, articleId=1203053368743781328, tenantId=1146029695717560320, journalId=1189873630562394117, language=CN, title=miR-149-5p靶向FGF21对大鼠心肌细胞的缺氧/复氧损伤的促进作用, columnId=1190310110472798614, journalTitle=解放军医学杂志, columnName=基础研究, runingTitle=null, highlight=null, articleAbstract=

目的 探究miR-149-5p与成纤维细胞生长因子21（FGF21）的靶向关系，及其对大鼠心肌细胞缺氧/复氧（H/R）损伤的影响。方法 将10只C56BL/6J雄性小鼠构建缺血再灌注（I/R）模型，qRT-PCR法检测I/R小鼠心脏组织中miR-149-5p表达水平的变化。将培养至生长对数期的H9c2大鼠心肌细胞分为对照组、H/R组、H/R+miR-149-5p inhibitor组、H/R+miR-149-5p NC组、H/R+miR-149-5p mimics组、H/R+miR-149-5p mimics+FGF21组，并诱导H/R模型及转染处理。ELISA法检测H9c2细胞中乳酸脱氢酶（LDH）和肌酸激酶（CK）的活性；MTT法检测细胞活力；流式细胞术检测细胞凋亡情况；qRT-PCR检测细胞中miR-149-5p的表达水平；Western blotting检测Bcl-2、Bax、cleaved caspase-3、FGF21蛋白的表达水平；双荧光素酶测定miR-149-5p与FGF21的靶向关系。结果 与假手术组比较，I/R小鼠心脏组织中miR-149-5p表达水平明显升高（P<0.05）。与对照组比较，H/R组H9c2细胞凋亡率、LDH和CK活性及Bax、cleaved caspase-3蛋白表达水平均明显升高（P<0.05），细胞活力及FGF21、Bcl-2蛋白表达水平明显降低（P<0.05）；与H/R组和H/R+miR-149-5p NC组比较，H/R+miR-149-5p inhibitor组细胞凋亡率、LDH和CK活性及Bax、cleaved caspase-3蛋白表达水平均明显降低（P<0.05），细胞活力及FGF21、Bcl-2蛋白表达水平均明显升高（P<0.05）；与H/R组比较，H/R+miR-149-5p mimics组细胞凋亡率、LDH和CK活性及Bax、cleaved caspase-3蛋白表达水平均明显升高（P<0.05），细胞活力及FGF21、Bcl-2蛋白表达水平均明显降低（P<0.05）。与H/R+miR-149-5p mimics组比较，H/R+miR-149-5p mimics+FGF21组细胞凋亡率及LDH和CK活性均明显降低（P<0.05），细胞活力明显升高（P<0.05）。双荧光素酶检测结果显示miR-149-5p与FGF21基因有靶向调控关系。结论 miR-149-5p在I/R小鼠心肌组织中明显上调，可负向调控FGF21水平，导致H/R处理的大鼠心肌细胞活力下降、凋亡增加。

, correspAuthors=张鹰, authorNote=null, correspAuthorsNote=, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=WfWFaMrjKxPu7GRPJCrZKQ==, magXml=Q8iPh5I2WQs5dpwIQVWPCw==, pdfUrl=null, pdf=YXFj/3ayYBD6qgE5OLIH+w==, pdfFileSize=1876680, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=cEiyRDWAQ5POPgS66vSGYw==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=9RS8OjI3cs31KneHWthrsw==, mapNumber=null, authorCompany=null, fund=null, authors=, authorsList=蒋先训, 张凯, 张鹰)}, authors=[Author(id=1203264515560415253, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, orderNo=0, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1203264515631718424, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, authorId=1203264515560415253, language=EN, stringName=Xian-Xun Jiang, firstName=Xian-Xun, middleName=null, lastName=Jiang, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=Department of Critical Care Medicine, the Second Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang, Hunan 421000, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1203264515703021593, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, authorId=1203264515560415253, language=CN, stringName=蒋先训, firstName=先训, middleName=null, lastName=蒋, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=南华大学衡阳医学院附属第二医院重症医学科，湖南衡阳 421000, bio={"content":"

蒋先训，硕士研究生，主治医师，主要从事心脏重症方面的基础研究

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蒋先训，硕士研究生，主治医师，主要从事心脏重症方面的基础研究

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I/R. 缺血再灌注；与假手术组比较，(1)P<0.05

, figureFileSmall=LxL+LGJ/cJvHnAh8H/feyw==, figureFileBig=IkeGMi1FQaI98eC67wq/nA==, tableContent=null), ArticleFig(id=1203264517087141934, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=EN, label=Fig. 2, caption=Effect of miR-149-5p on cell viability of H9c2 rat's cardiomyocytes, figureFileSmall=Ia1MacRdZx1LLS+hfUkwnw==, figureFileBig=rHwJRMQ618JBE+vjHhpoWA==, tableContent=null), ArticleFig(id=1203264517145862191, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=CN, label=图2, caption=miR-149-5p对H9c2大鼠心肌细胞活力的影响

H/R. 缺氧/复氧；与对照组比较，(1)P<0.05；与H/R组比较，(2)P<0.05；与H/R+miR-149-5p NC组比较，(3)P<0.05

, figureFileSmall=Ia1MacRdZx1LLS+hfUkwnw==, figureFileBig=rHwJRMQ618JBE+vjHhpoWA==, tableContent=null), ArticleFig(id=1203264517204582448, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=EN, label=Fig. 3, caption=Effect of miR-149-5p on activity of LDH and CK in H9c2 rat's cardiomyocytes, figureFileSmall=WfPis1wD63UhPdb+OU/lFA==, figureFileBig=HtM5niAPvXfQMYQTnZxWsg==, tableContent=null), ArticleFig(id=1203264517263302705, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=CN, label=图3, caption=miR-149-5p对H9c2大鼠心肌细胞中LDH和CK活性的影响

H/R. 缺氧/复氧；LDH. 乳酸脱氢酶；CK. 肌酸激酶；与对照组比较，(1)P<0.05；与H/R组比较，(2)P<0.05；与H/R+miR-149-5p NC组比较，(3)P<0.05

, figureFileSmall=WfPis1wD63UhPdb+OU/lFA==, figureFileBig=HtM5niAPvXfQMYQTnZxWsg==, tableContent=null), ArticleFig(id=1203264517322022962, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=EN, label=Fig. 4, caption=Effect of miR-149-5p on apoptosis of H9c2 rat's cardiomyocytes, figureFileSmall=4RDyO460XLuLVob54446Cw==, figureFileBig=2gSRgLm69XeV9QpppnE1dQ==, tableContent=null), ArticleFig(id=1203264517405909043, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=CN, label=图4, caption=miR-149-5p对H9c2大鼠心肌细胞凋亡的影响

H/R. 缺氧/复氧；与对照组比较，(1)P<0.05；与H/R组相比，(2)P<0.05；与H/R+miR-149-5p NC组比较，(3)P<0.05

, figureFileSmall=4RDyO460XLuLVob54446Cw==, figureFileBig=2gSRgLm69XeV9QpppnE1dQ==, tableContent=null), ArticleFig(id=1203264517489795124, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=EN, label=Fig. 5, caption=Effect of miR-149-5p on expression of apoptosis-related proteins in H9c2 rat's cardiomyocytes (Western blotting), figureFileSmall=36S8uRm/Ddodpn6t1vOhzg==, figureFileBig=EPVsI0YZI/yNYvakrb7Hsw==, tableContent=null), ArticleFig(id=1203264517569486901, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=CN, label=图5, caption=miR-149-5p对H9c2大鼠心肌细胞凋亡相关蛋白表达的影响

H/R. 缺氧/复氧；与对照组比较，(1)P<0.05；与H/R组相比，(2)P<0.05；与H/R+miR-149-5p NC组比较，(3)P<0.05

, figureFileSmall=36S8uRm/Ddodpn6t1vOhzg==, figureFileBig=EPVsI0YZI/yNYvakrb7Hsw==, tableContent=null), ArticleFig(id=1203264517670150198, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=EN, label=Fig. 6, caption=Prediction of binding sites on FGF21 3' UTR with miR-149-5p and the results of dual-luciferase reporter assay, figureFileSmall=a755wxS3+AphKa/DHtNCVw==, figureFileBig=ZZdSb0BLTSM8YmR6mx0VeQ==, tableContent=null), ArticleFig(id=1203264517745647671, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=CN, label=图6, caption=miR-149-5p与FGF21 3' UTR的预测结合位点及双荧光素酶检测结果

WT. 野生型；MUT. 突变型；UTR. 非翻译区；A. 生物信息学预测的结合位点；B. 荧光素酶检测结果；与miR-149-5p NC+FGF21 WT组比较，(1)P<0.05

, figureFileSmall=a755wxS3+AphKa/DHtNCVw==, figureFileBig=ZZdSb0BLTSM8YmR6mx0VeQ==, tableContent=null), ArticleFig(id=1203264517808562232, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=EN, label=Fig. 7, caption=Effect of miR-149-5p on expression of FGF21 in H9c2 rat's cardiomyocytes (Western blotting), figureFileSmall=9OQ7+qn47kkvS9j17yqr5Q==, figureFileBig=RoKxpQ1t2n03cjpt8zNftQ==, tableContent=null), ArticleFig(id=1203264517867282489, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=CN, label=图7, caption=miR-149-5p对H9c2大鼠心肌细胞FGF21蛋白表达的影响

H/R. 缺氧/复氧；与对照组比较，(1)P<0.05；与H/R组比较，(2)P<0.05；与H/R+miR-149-5p NC组比较，(3)P<0.05

, figureFileSmall=9OQ7+qn47kkvS9j17yqr5Q==, figureFileBig=RoKxpQ1t2n03cjpt8zNftQ==, tableContent=null), ArticleFig(id=1203264517934391354, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=EN, label=Fig. 8, caption=Effect of miR-149-5p on activity of LDH and CK in H9c2 rat's cardiomyocytes through FGF21, figureFileSmall=qEKuqvDSI8leUGxKN2HzfQ==, figureFileBig=hGTZlXV1aFl1adGON99T6A==, tableContent=null), ArticleFig(id=1203264519054270523, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=CN, label=图8, caption=miR-149-5p通过FGF21对H9c2大鼠心肌细胞中LDH和CK活性的影响

H/R. 缺氧/复氧；LDH. 乳酸脱氢酶；CK. 肌酸激酶；与H/R+miR-149-5p mimics组比较，(1)P<0.05

, figureFileSmall=qEKuqvDSI8leUGxKN2HzfQ==, figureFileBig=hGTZlXV1aFl1adGON99T6A==, tableContent=null), ArticleFig(id=1203264519133962300, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=EN, label=Tab. 1, caption=

Primer sequences for qRT-PCR

, figureFileSmall=null, figureFileBig=null, tableContent=

基因	引物序列
miR-149-5p	正义：5'-TCTGGCTCCGTGTCTTCACT-3'
	反义：5'-TCTGGCTCCGTGTCTTCACTCCC-3'
U6	正义：5'-TGGAACGCTTCACGAATTTGCG-3'
	反义：5'-GGAACGATACAGAGAAGATTAGC-3'

), ArticleFig(id=1203264519209459773, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1203053368743781328, language=CN, label=表1, caption=

qRT-PCR引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=

基因	引物序列
miR-149-5p	正义：5'-TCTGGCTCCGTGTCTTCACT-3'
	反义：5'-TCTGGCTCCGTGTCTTCACTCCC-3'
U6	正义：5'-TGGAACGCTTCACGAATTTGCG-3'
	反义：5'-GGAACGATACAGAGAAGATTAGC-3'

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