口腔医学研究

口腔医学研究 ›› 2025, Vol. 41 ›› Issue (12): 1074-1080.DOI: 10.13701/j.cnki.kqyxyj.2025.12.011

• 口腔生物学研究 • 上一篇    下一篇

TGF-β1/Smad7对大鼠牙胚上皮样细胞光蛋白聚糖mRNA表达的影响

崔颖颖1,2, 孙轶群2, 张斌1,3, 刘贞2, 田剑刚3, 黄瑞哲1,3*   

  1. 1.西安交通大学口腔医院 陕西省颅颌面精准医学研究重点实验室 陕西 西安 710004;
    2.滨州医学院附属医院儿童口腔科 山东 滨州 256600;
    3.西安交通大学口腔医院口腔预防保健中心 陕西 西安 710004
  • 收稿日期:2025-03-17 出版日期:2025-12-28 发布日期:2025-12-23
  • 通讯作者: *黄瑞哲,E-mail:huangrzh@mail.xjtu.edu.cn
  • 作者简介:崔颖颖(1986~ ),女,山东利津人,硕士,主治医师,研究方向:儿童口腔相关疾病的临床研究。
  • 基金资助:
    陕西省自然科学基础研究计划项目(编号:2023-JC-QN-0980)

Effect of TGF-β1/Smad7 on Lumican mRNA Expression in Tooth Germ Epithelioid Cells of Rats

CUI Yingying1,2, SUN Yiqun2, ZHANG Bin1,3, LIU Zhen2, TIAN Jiangang3, HUANG Ruizhe1,3*   

  1. 1. Key Lab of Craniomaxillofacial Precision Medicine Research in Shaanxi Province, Hospital of Stomatology Xi'an Jiaotong University, Xi'an 710004,China;
    2. Department of Pediatric Dentistry, Hospital Affiliated to Binzhou Medical University, Binzhou 256600, China;
    3. Oral Preventive Health Care Center, Hospital of Stomatology Xi'an Jiaotong University, Xi'an 710004, China
  • Received:2025-03-17 Online:2025-12-28 Published:2025-12-23

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摘要： 目的: 通过原代细胞培养探究转化生长因子(transforming growth factor-β1,TGF-β1)/Smad7信号通路对光蛋白聚糖表达的影响。方法: 对SD大鼠牙胚上皮样细胞进行体外分离培养、纯化、鉴定。选择生长良好的体外培养SD大鼠牙胚细胞作为研究对象,采用实时荧光定量逆转录聚合酶链反应检测细胞中Smad7基因mRNA表达情况。探究电转染RNA干扰对Smad7基因表达的影响。给予不同浓度TGF-β1(0、0.1和10 ng/mL)对细胞光蛋白聚糖mRNA表达的影响。结果: 酶消化法对SD大鼠牙胚上皮样细胞进行原代培养,显微镜下显示细胞形态一致,呈圆形或椭圆形,单核且胞浆丰富。免疫组织化学染色结果显示,上皮样细胞中角蛋白14染色呈阳性,波形丝蛋白染色呈阴性,证明所得细胞为上皮样细胞。实时荧光定量聚合酶链反应证明SD大鼠牙胚上皮样细胞中Smad7基因表达,但低于GAPDH表达。电穿孔转染结果显示,干扰片段pRNAi-Smad1949干扰后,Samd7表达降低。与对照组相比,10 ng/mL的TGF-β1处理细胞中光蛋白聚糖mRNA表达明显升高(P<0.05),表明TGF-β1增加光蛋白聚基因mRNA表达。pRNAi-Smad1949转染抑制Samd7后,光蛋白聚糖mRNA表达升高。结论: TGF-β1通过激活Samd7信号,上调牙胚上皮细胞中光蛋白聚糖mRNA表达,且具有剂量依赖性。这可能影响上皮-间质相互作用,进而影响牙齿的形成与分化。

关键词: 转化生长因子β, Smad7, 光蛋白聚糖

Abstract: Objective: To investigate the effect of the transforming growth factor-β1 (TGF-β1)/Smad7 signaling pathway on lumican expression in primary cultured cells. Methods: Epithelioid cells from the tooth germs of SD rats were isolated, cultured, purified, and identified in vitro. Well-grown cells were selected for study. Smad7 mRNA expression was detected using quantitative real-time reverse transcription PCR (qRT-PCR). RNA interference via electroporation was employed to explore its impact on Smad7 expression. Cells were treated with varying concentrations of TGF-β1 (0, 0.1, and 10 ng/mL) to assess its effect on lumican mRNA expression. Results: Primary culture of SD rat tooth germ epithelioid cells was established via enzymatic digestion. Microscopic observation revealed cells with homogeneous morphology, exhibiting round or oval shapes, single nuclei, and abundant cytoplasm. Immunohistochemical staining confirmed the epithelial origin of the cells: cytokeratin 14 (epithelial marker) was positively expressed, while vimentin (mesenchymal marker) was negative. qRT-PCR demonstrated detectable Smad7 mRNA expression in these cells, though at lower levels compared to GAPDH. Electroporation-based transfection with the pRNAi-Smad1949 interference fragment significantly downregulated Smad7 expression. Compared to the control group, treatment with 10 ng/mL TGF-β1 markedly increased lumican mRNA expression (P<0.05), indicating TGF-β1 enhanced lumican transcription. Intriguingly, suppression of Smad7 via pRNAi-Smad1949 transfection also led to elevated lumican mRNA expression. Conclusion: TGF-β1 upregulates lumican mRNA expression in tooth germ epithelioid cells by activating Smad7 signaling, with a dose-dependent effect. This mechanism may influence epithelial-mesenchymal interactions, thereby affecting tooth formation and differentiation.

Key words: transforming growth factor-β, Smad7, lumican