TGF―β1调控ERKMAPK信号通路对人牙髓细胞增殖和分化能力影响研究.docVIP

TGF―β1调控ERKMAPK信号通路对人牙髓细胞增殖和分化能力影响研究 　　[摘要]目的：研究TGF-β1?ρ浪柘赴?增殖、分化和ERK/MAPK信号通路的影响。方法：采用Western blot检测TGF-β1在急性牙髓组织、慢性牙髓组织和正常牙髓组织中的表达，然后采用Ⅰ型胶原酶消化分离培养牙髓细胞，不同浓度的TGF-β1处理牙髓细胞7d，CCK-8法检测细胞增殖。采用TGF-β1（5.0μg/L）和U0126（10μmol/L）分别单独或联合作用于牙髓细胞7d，CKK-8法检测细胞增殖；在第1、3、5和7天检测碱性磷酸化酶活性；第7天时Western blot检测ERK/MAPK信号通路中的关键蛋白PPAR-γ和ELK-1水平。结果：TGF-β1在牙髓炎组织中低表达，且TGF-β1浓度在0.1～5.0?g/L时，从作用第3天开始能够显著促进牙髓细胞的增殖（P0.05），具有时间和剂量依赖效应。U0126能够抑制牙髓细胞的增殖和碱性磷酸化酶的活性，具有时间依赖性。TGF-β1能够促进PPAR-γ和ELK-1蛋白的表达，而U0126能够降低TGF-β1对PPAR-γ和ELK-1蛋白表达的促进作用。结论：TGF-β1可以促进牙髓细胞的增殖和分化，且具有时间和浓度依赖效应，其作用机制可能与ERK/MAPK信号通路相关。 　　[关键词]牙髓细胞；增殖；分化；碱性磷酸化酶；信号通路 　　[中图分类号]R781.3 [文献标志码]A [文章编号]1008-6455（2018）04-0094-04 　　Effect of TGF-β1 on the Proliferation and Differentiation of Human Dental Pulp Cell Based on ERK/MAPK Signaling Pathway 　　XU Wan-tian，CHEN Yun，LIN Nan，TANG Yu-xiang，YAN Xiang 　　（The Third Clinic Department，Nanjing Stomatological Hospital Medical School of Nanjing University，Nanjing 210008，Jiangsu，China） 　　Abstract： Objective To study the effects of TGF-β1 on the proliferation， differentiation and ERK/MAPK signaling pathway of human dental pulp cells. Methods We used Western blot to detect TGF-β1 expression in acute and chronic dental pulp tissue organization and normal dental pulp tissue. Primary human dental pulp cells were isolated and cultured by typeⅠcollagenase digestion. Human dental pulp cells were treated by different concentrations of TGF-β1 for 7d. Used CCK-8 method to detect cell proliferation. Human dental pulp cells were treated by TGF-β1 （5.0μg/L） and U0126 （10μmol/L） separately or jointly for 7d and cell proliferation was detected by CKK-8 method. The activity of alkaline phosphorylase was detected in 1d， 3d， 5d and 7d. The levels of key proteins PPAR-γ and ELK-1 in the ERK/MAPK signaling pathway were detected by Western blot. Results TGF-β1 was low expression in the pulpitis tissue， and TGF-β1 concentration at 0.1-5.0μg/L， the proliferation of pulp cells was significantly promoted from the first 3d