BMP2和BMP4对牙釉质基质蛋白酶基因表达的调控

›› 2020, Vol. 40 ›› Issue (9): 773-777.

• 基础研究 • 下一篇

BMP2和BMP4对牙釉质基质蛋白酶基因表达的调控

刘向晖¹,耿硕硕¹,吴迪¹,李莹¹,罗晓娜¹,许莹莹¹,袁浩泽¹,王秀梅²,谢晓华¹

1. 哈尔滨医科大学附属第二医院
2. 哈尔滨市哈尔滨医科大学附属第二医院口腔内科

收稿日期:2020-03-03 修回日期:2020-04-04 出版日期:2020-09-28 发布日期:2020-10-07
通讯作者: 谢晓华 E-mail:xiexiaohua_79@126.com
基金资助:
国家自然科学基金;黑龙江省自然基金重点项目

Regulation of protease gene expression in enamel matrix by BMP2 and BMP4

Received:2020-03-03 Revised:2020-04-04 Online:2020-09-28 Published:2020-10-07

摘要/Abstract

摘要： 目的　研究BMP2和BMP4对小鼠ALC（ameloblast-lineage cell）内牙釉质基质蛋白酶中基质金属蛋白酶20（matrix metalloproteinase 20，MMP20）和激肽释放酶4（kallikrein 4，KLK4）表达中的调控作用。方法　在ALC细胞系中加入BMP2和BMP4重组蛋白，利用免疫印迹试验和荧光定量PCR检测BMP通路信号分子和牙釉质基质蛋白酶的表达变化。同期，在另一组ALC细胞系中加入BMP通路抑制剂（Dorsomorphin，Dor）,检测牙釉质基质蛋白酶的表达情况。结果　在BMP2和BMP4蛋白的作用下，ALC细胞中BMP通路的经典通路（Smad1/5/8）和非经典通路（p38）的表达显著增强，同时MMP20和KLK4的基因表达也有明显上调。同期添加Dorsomorphin的ALC细胞中BMP通路的经典通路的表达明显下降，而非经典通路的表达变化不大，同时MMP20和KLK4的基因表达也有明显的下调趋势。在另一组实验中添加BMP抑制剂的处理组则表现出MMP20和KLK4蛋白表达的下调，而使用BMP2和BMP4对抑制剂组进行处理的挽救组则表现出MMP20和KLK4蛋白水平的明显恢复。结论　BMP2和BMP4可以通过影响BMP信号经典通路对牙釉质形成过程中牙釉质基质蛋白酶的表达具有调控作用。

关键词: BMP2, BMP4, MMP20, KLK4

Abstract: Abstract: Objective To investigate the effects of BMP2 and BMP4 on the expression of matrix metalloproteinase 20 (MMP20) and kallikrein 4 (KLK4) in ALC (ameloblast-lineage cell). Methods Recombinant proteins of BMP2 and BMP4 were added to ALC cell lines, and changes in BMP pathway signaling and enamel matrix protease expression were detected by Western blot and real-time PCR. At the same time, another group of cells was treated with a BMP pathway inhibitor（Dorsomorphin, Dor）to detect the expression of enamel matrix proteases. Results Under the action of BMP2 and BMP4 proteins, the expression of classical pathway（Smad1/5/8）and non-classical pathway（p38）of BMP pathway in ALC cells was significantly enhanced, and the expression of MMP20 and KLK4 genes was also significantly up-regulated. In the same period, the expression of the classical pathway of BMP pathway in ALC cells with Dorsomorphin was significantly decreased, while the expression of the non-classical pathway was not significantly changed, and the expression of MMP20 and KLK4 genes also showed a significant down-regulation trend. In another set of experiments, the treatment group with BMP inhibitor showed down-regulation of MMP20 and KLK4 protein expression, while the rescue group treated with BMP2 and BMP4 showed significant recovery of MMP20 and KLK4 protein levels. Conclusions BMP2 and BMP4 can regulate the expression of enamel matrix protease during tooth enamel formation by influencing the BMP classical pathway.

Key words: bone morphogenetic protein2, bone morphogenetic protein4, metalloproteinase 20, kallikrein 4

中图分类号:

Q753

刘向晖耿硕硕吴迪李莹罗晓娜许莹莹袁浩泽王秀梅谢晓华. BMP2和BMP4对牙釉质基质蛋白酶基因表达的调控[J]. 口腔医学, 2020, 40(9): 773-777.

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BMP2和BMP4对牙釉质基质蛋白酶基因表达的调控

Regulation of protease gene expression in enamel matrix by BMP2 and BMP4

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