PAD4在颌骨生长发育调控中的作用初探

doi:10.13591/j.cnki.kqyx.2025.04.001

摘要/Abstract

摘要：

目的探究肽基精氨酸脱亚氨酶4(peptidyl arginine deiminase 4,PAD4)对颌骨间充质干细胞(mesenchymal stem cells isolated from oral bones,OMSC)分化及颅颌面发育的影响。方法通过免疫荧光技术检测PAD4在胚胎发育期小鼠颌骨中的表达,构建PAD4基因敲除(PAD4-KO)小鼠模型,通过micro CT比较PAD4-KO小鼠与野生型小鼠颌骨发育差异;通过CCK-8、Transwell等实验测定PAD4-KO与野生型(wild type,WT)小鼠OMSC增殖及迁移能力的差异,通过ALP实验测定了两组小鼠OMSC的成骨能力,并通过免疫荧光、PCR测定成骨相关基因的表达差异。结果 PAD4在小鼠E13.5期胚胎颌骨组织中表达,细胞水平上PAD4在OMSC细胞核及线粒体中表达。观察见PAD4-KO小鼠下颌骨发育迟缓,micro CT示PAD4-KO小鼠颅骨及颌骨发育迟缓、矿化不良。细胞实验示PAD4-KO小鼠OMSC细胞增殖及迁移能力下降,ALP实验结果示OMSC成骨矿化能力下降,免疫荧光和PCR结果示成骨相关基因表达水平降低。此外研究发现PAD4可能通过调节Runx2的转录影响OMSC的矿化。结论 PAD4在胚胎发育期颌骨中表达,其可能通过调节OMSC的分化功能,从而影响胚胎发育,导致颅颌面发育异常。

关键词: 肽基精氨酸脱亚氨酶4, 颌骨间充质干细胞, 颅颌面发育

Abstract:

Objective To investigate the effect of peptidyl arginine deiminase 4(PAD4) on the differentiation of mesenchymal stem cells isolated from oral bones(OMSC) and craniomaxillofacial development. Methods Immunofluorescence was used to detect the expression of PAD4 in the mandibular of mice E13.5 embryo. A peptidyl arginine deiminase 4 knockout(PAD4-KO) mouse model was constructed. Craniomaxillofacial development was investigated by micro-CT. CCK-8 assay and Transwell assay were used to detect the OMSC proliferation ability and migration ability of PAD4-KO and wild type (WT) mouse. ALP staining was used to detect the changes in OMSC osteogenic differentiation ability. The expression of osteogenesis-related genes was detected by immunofluorescence and PCR assay. Results PAD4 was highly expressed in the mandibular tissue of mouse embryos at E13.5. On the cellular level, PAD4 was expressed in the nucleus and mitochondria of OMSC. Compared to the WT mice,micro-CT showed that PAD4-KO mice had retrusive jaw and decreased mineralization. The proliferation and migration ability of OMSC in PAD4-KO mice were decreased. OMSCs lacking PAD4 had significantly decreased ALP staining level, and the expression levels of osteogenesis-related genes were decreased. In addition, it was found that PAD4 might affect OMSC mineralization by regulating Runx2 transcription. Conclusion PAD4 is expressed in the jaw during embryonic development. It might affect the embryonic development by regulating the proliferation and differentiation of OMSC, leading to craniomaxillofacial abnormalities

Key words: peptidyl arginine deiminase 4, mesenchymal stem cells isolated from oral bones, craniomaxillofacial development

中图分类号:

R783.5

闫行之, 蔡新誉, 陈思麦, 方韦文, 雷凡, 曹丹, 张阳. PAD4在颌骨生长发育调控中的作用初探[J]. 口腔医学, 2025, 45(4): 241-247.

YAN Xingzhi, CAI Xinyu, CHEN Simai, FANG Weiwen, LEI Fan, CAO Dan, ZHANG Yang. Preliminary study on the role of peptidyl arginine deiminase 4 in the regulation of maxillofacial development[J]. Stomatology, 2025, 45(4): 241-247.

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