摘要
成纤维细胞生长因子20(fibroblast growth factor 20,FGF20)在心肌肥大中具有保护作用,但其作用机制尚未明确。因此,该研究旨在探究FGF20改善心肌细胞肥大的分子作用机制。该研究使用10μmol/L异丙肾上腺素(isoproterenol,ISO)刺激原代心肌细胞NRCMs 48 h,构建心肌细胞肥大模型,并同时对NRCMs分别给予100 ng/mL重组蛋白FGF20以及50 ng/mL重组蛋白骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)共处理48 h,并借助转录组测序分析FGF20处理前后的差异表达基因(differential expression genes,DEGs),再对其进行GO和KEGG富集分析以及PPI网络构建。使用机器学习算法LASSO和Random Forest筛选得出核心差异基因(Core-DEGs),并对其进行ROC分析。最后,使用蛋白质免疫印迹以及RT-qPCR验证核心差异基因的表达,TUNEL染色实验检测心肌细胞凋亡情况。结果表明,FGF20可抑制ISO诱导的心肌肥大指标ANP、BNP表达上调,证实FGF20具有心肌细胞肥大保护作用。转录组测序分析发现,与ISO组相比,FGF20处理后共鉴定出42个差异表达基因。GO和KEGG富集分析结果表明,差异表达基因主要富集在多个免疫调控以及凋亡信号通路。2种机器学习算法联合筛选后发现Bmp4为Core-DEGs,且其AUC为1。实验结果表明,FGF20可抑制ISO诱导的BMP4表达上调以及心肌细胞凋亡;而BMP4激活后FGF20心肌细胞肥大保护作用被明显削弱,并伴随ANP、BNP蛋白水平上调以及心肌细胞凋亡增多。总之,FGF20通过抑制BMP4缓解心肌细胞凋亡进而改善心肌细胞肥大。
FGF20(fibroblast growth factor 20)has a protective role in cardiac hypertrophy,but its molecu-lar mechanism still unclear.Therefore,this study is aimed to explore the molecular mechanisms of FGF20 protecting against cardiomyocyte hypertrophy.NRCMs(neonatal rat cardiomyocytes)were stimulated with 10μmol/L ISO(iso-proterenol)for 48 h to establish a cardiomyocyte hypertrophy model,and then co-treated with 100 ng/mL recom-binant protein FGF20 or 50 ng/mL recombinant protein BMP4(bone morphogenetic protein 4)for 48 h.Transcrip-tome was performed to identify DEGs(differential expression genes)between ISO group and ISO+FGF20 group.DEGs were subjected to GO,KEGG enrichment analysis and PPI(protein-protein interaction)network construc-tion.Machine learning algorithms were employed to identify Core-DEGs(core differentially expressed genes),fol-lowed by ROC(receiver operating characteristic)analysis.Finally,the expression of Core-DEGs was validated by Western blot and RT-qPCR analysis,and TUNEL staining was employed to detect cardiomyocyte apoptosis.The re-sult showed that FGF20 alleviated ISO-induced cardiomyocyte hypertrophy,evidenced by decreased levels of ANP and BNP proteins.Transcriptome analysis identified 42 DEGs after FGF20 treatment compared to ISO stimulation.GO and KEGG enrichment analyses revealed that 42 DEGs were associated with multiple immune regulation and apoptosis signaling pathways.Notably,Bmp4 was identified as the Core-DEG by two classic machine learning al-gorithms(LASSO and Random Forest)and its AUC value was 1.Furthermore,FGF20 inhibited BMP4 expression and cardiomyocyte apoptosis induced by ISO.However,activation of BMP4 counteracted the protection of FGF20 against myocardial hypertrophy,resulting in increased levels of ANP and BNP proteins and cardiomyocyte apopto-sis.In summary,FGF20 exerts a protective effect on cardiomyocyte hypertrophy by suppressing apoptosis through the inhibition of BMP4.
作者
梅淋
王旭
陈慧楠
周旋
陈云杰
MEI Lin;WANG Xu;CHEN Huinan;ZHOU Xuan;CHEN Yunjie(Department of Pharmacy,Xiamen Medical College,Xiamen 361000,China;School of Pharmaceutical Science,Wenzhou Medical University,Wenzhou 325035,China;Department of Pharmacy,the First Affiliated Hospital of Ningbo University,Ningbo 315010,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2024年第7期1335-1346,共12页
Chinese Journal of Cell Biology
基金
浙江省自然科学基金(批准号:LQ24H020001)
国家自然科学基金(批准号:82170365)
厦门市自然科学基金(批准号:3502Z202372061)
宁波市自然科学基金(批准号:2022J223)资助的课题。
