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SOX9与Smad4在不同角膜基质细胞中的表达及相互作用

陈冬梅 华耘 刘佳琳 赵新荣

眼科新进展2026,Vol.46Issue(4):269-274,6.
眼科新进展2026,Vol.46Issue(4):269-274,6.DOI:10.13389/j.cnki.rao.2026.0047

SOX9与Smad4在不同角膜基质细胞中的表达及相互作用

Expression and interaction of SRY-box transcription factor 9 and Smad family member 4 in different keratocytes

陈冬梅 1华耘 2刘佳琳 1赵新荣1

作者信息

  • 1. 832008 新疆维吾尔自治区石河子市,石河子大学第一附属医院
  • 2. 610041 四川省成都市,四川大学华西医院
  • 折叠

摘要

Abstract

Objective To investigate the differential expression and interaction of SRY-box transcription factor 9(SOX9)and Smad family member 4(Smad4)in resting-state keratocytes(KCs),corneal fibroblastic cells(CFs),and cor-neal myofibroblasts(CMFs),and to provide experimental evidence for the mechanism and targeted therapy of corneal fi-brotic disorders.Methods Bioinformatic methods were used to analyze the GSE287915 dataset in the Gene Expression Omnibus database,screen for differentially expressed genes,and focus on SOX9.Quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR)was performed to verify the expression of SOX9 in the mouse KCs(m-KCs),mouse CFs(m-CFs),and mouse CMFs(m-CMFs).The Gene Transcription Regulation Database(JASPAR)was used to predict the potential binding sites between SOX9 and Smad4.qRT-PCR was applied to detect the expression levels of Smad4 in the said three types of cells,and finally,co-immunoprecipitation(Co-IP)was used to verify the interaction between SOX9 and Smad4 proteins.Results Bioinformatic analysis showed that compared with KCs,SOX9 was significantly highly expressed in CFs and CMFs(both P<0.05).The qRT-PCR detection results revealed that the expression levels of Smad4 and SOX9 followed the trend:m-KCs group<m-CFs group<m-CMFs group,with statistically significant differences among the three groups(all P<0.001).The JASPAR database predicted that there were potential binding sites between SOX9 and the Smad4 promoter region.Co-IP results demonstrated that when SOX9 and Smad4 antibodies were used as the capture antibodies,the binding amount of Smad4 and SOX9 in the m-CMFs group was significantly higher than that in the m-CFs group and the m-KCs group(both P<0.05),while no obvious binding was detected in the m-KCs group.Conclusion The expression lev-els of SOX9 and Smad4 in in vitro cultured m-KCs,m-CFs,and m-CMFs are elevated with the degree of cellular activation and fibrosis progression.Moreover,a specific interaction between them has been identified in fibrotic phenotype m-CFs and m-CMFs induced in vitro.The upregulation of their expression and their protein-protein interaction may contribute to the fi-brosisn-related activation of keratocytes in vitro,providing an experimental basis for further mechanism studies on corneal fibrotic diseases.

关键词

角膜基质细胞/角膜成纤维细胞/角膜肌成纤维细胞/性别决定区Y相关高迁移率族盒蛋白9/Smad家族成员4/相互作用

Key words

keratocytes/corneal fibroblasts/corneal myofibroblasts/SRY-box transcription factor 9/Smad family member 4/interaction

分类

医药卫生

引用本文复制引用

陈冬梅,华耘,刘佳琳,赵新荣..SOX9与Smad4在不同角膜基质细胞中的表达及相互作用[J].眼科新进展,2026,46(4):269-274,6.

基金项目

国家自然科学基金项目(编号:82060171) (编号:82060171)

眼科新进展

1003-5141

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