TAZ在张应力促进人牙周膜细胞骨向分化中的作用

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摘要目的：探索PDZ结合基序转录共激活因子（TAZ）参与张应力促进间充质干细胞（MSCs）骨向分化的调控作用及分子机制。方法：原代培养人牙周膜细胞（hPDLCs）。利用Flexcell系统对hPDLCs体外加载12%，0.5 Hz张应力，实时荧光定量PCR及Western blot检测hPDLCs中TAZ及成骨标志物核心结合因子α1（Cbfα1）、I型胶原（COL I）、骨钙素（OCN）、锌指结构转录因子（OSX）的表达变化；蛋白质免疫共沉淀检测TAZ与Cbfα1的结合情况。结果：加载12%，0.5 Hz张应力24 h及48 h时显著促进hPDLCs的TAZ、Cbfα1、COL I、OSX、OCN mRNA及蛋白表达水平（P＜0.05）。蛋白质免疫共沉淀证实TAZ与Cbfα1的结合量显著增加。hPDLCs转染TAZ siRNA 72 h，加载12%，0.5 Hz张应力24 h，TAZ蛋白水平及张应力所促进的细胞成骨蛋白表达下降。结论：张应力作用下TAZ通过上调Cbfα1的表达与转录活性诱导hPDLCs骨向分化。

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	陆素文
	胡碧波
	邓辉

关键词 ： TAZ, 张应力, 核心结合因子&alpha, 1, 人牙周膜细胞, 骨向分化

Abstract：Objective: To explore the role of PDZ binding transcriptional activator of transcription (TAZ) in tensile stress-induced cytoskeleton changes of human periodontal ligament cells (hPDLCs) to promote osteogenic differentiation. Methods: hPDLCs were obtained from PDL tissues in intact extracted third molars. Flexcell system was applied to construct the tensile stress loading model in vitro. Changes in TAZ and osteoblastic markers Cbfα1, collagen I (COL I), osteocalcin (OCN) and osterix (OSX) in hPDLCs were detected by Western blot and PCR. And the connection between TAZ and Cbfα1 was detected by co-immunoprecipitation. TAZ siRNA transfection was used to determine the role of TAZ in bone differentiation induced by tensile stress. Results: TAZ, Cbfα1, COL I, OSX and OCN mRNA and protein levels increased after 0.5 Hz and 12% tensile stress was applied to hPDLCs for 24 h and 48 h. There was a much more obvious connection between TAZ and Cbfα1 under tension for 24 h, compared with the control group. TAZ decreased after TAZ siRNA transfection in hPDLCs. Western blot results indicated that Cbfα1, COL I, OSX and OCN increased by tension were down-regulated after TAZ siRNA interference. Conclusion: TAZ induces osteogenic differentiation of human periodontal ligament cells by up-regulating Cbfα1 expression and transcriptional activity during tensile stress.

Key words： TAZ tensile stress core binding factor α1 human periodontal ligament cells osteogenic differentiation

收稿日期: 2019-04-29

基金资助:浙江省自然科学基金资助项目（LY17H140009）；浙江省医药卫生科技计划项目（LGF19H140001）。

通讯作者: 邓辉，主任医师，硕士生导师，Email：dh0726@163.com。

作者简介: 陆素文（1984-），女，浙江宁波人，在职硕士生。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2019/V49/I9/634

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