内皮素受体A对牙周膜干细胞骨向分化能力影响的研究

›› 2020, Vol. 11 ›› Issue (3): 161-163.

内皮素受体A对牙周膜干细胞骨向分化能力影响的研究

梁莉¹,许亦权¹,杨楠²,王辰¹

1. 解放军总医院第八医学中心口腔科
2. 解放军第309医院口腔科

收稿日期:2020-05-06 修回日期:2020-07-08 出版日期:2020-09-25 发布日期:2020-09-30
通讯作者: 梁莉 E-mail:lily1981125@126.com
基金资助:
国家自然科学基金

Effects of endothelin receptor A on osteoblastic differentiation function of periodontal ligament stem cells

Received:2020-05-06 Revised:2020-07-08 Online:2020-09-25 Published:2020-09-30

摘要/Abstract

摘要： 目的：研究内皮素受体A（ETRA）对牙周膜干细胞（PDLSCs）成骨分化能力的影响。方法：构建ETRA小干扰RNA（siRNA）载体，转染PDLSCs后，采用实时定量RT-PCR及Western?Blot检测转染前后ETRA的表达情况。用100?nmol/L内皮素-1（ET-1）刺激siETRA转染和未转染的的PDLSCs?14?d，碱性磷酸酶（ALP）染色观察PDLSCs成骨能力的变化，实时定量RT-PCR及Western?Blot检测PDLSCs?Runt相关转录因子2（RUNX2）和骨钙素（OCN）的表达情况。结果：ETRA干扰载体可有效地抑制PDLSCs中ETRA表达。ET-1刺激后，未转染组PDLSCs?ALP染色增强，RUNX2和OCN表达升高（P＜0.05），但对siETRA稳定转染组ALP染色、RUNX2和OCN表达无显著影响（P＞0.05）。结论：ET-1可能通过ETRA促进PDLSCs的骨向分化能力。

关键词: 内皮素受体A, 牙周膜干细胞, 成骨分化, 小干扰 RNA

Abstract: Objective:?To investigate the effects of endothelin receptor A（ETRA）on osteogenic differentiation function of periodontal ligament stem cells (PDLSCs). Methods:?The small interfering RNA (siRNA) technique was employed to inhibit ETRA expression in PDLSCs and cells were cultured with ET-1 (100?nmol/L) for 14 days. The osteoblastic differentiation function of PDLSCs was observed by alkaline phosphatase staining. The expression of RUNX2 and OCN mRNAs and proteins in PDLSCs were detected by real-time RT-PCR and Western Blot, respectively. Results:?ETRA interference vector can effectively inhibit the expression of ETRA in PDLSCs. ET-1 significantly enhanced the expression of RUNX2 and OCN of PDLSCs（P＜0.05). However, ET-1 had no effect on the expression of RUNX2 and OCN of PDLSCs stably transfected by siETRA（P＞0.05）. Conclusions: These results indicate that endothelin-1 may stimulate the osteogenic differentiation of PDLSCs via ETRA.

Key words: endothelin receptor A, periodontal ligament stem cells, small interfering RNA

梁莉许亦权杨楠王辰. 内皮素受体A对牙周膜干细胞骨向分化能力影响的研究[J]. 口腔生物医学, 2020, 11(3): 161-163.

参考文献 14

[1]	Seo BM, Miura M, Gronthos S, et al.Investigation of multipotent postnatal stem cells from human periodontal ligament[J].Lancet, 2004, 364(9429):149-155
[2]	Tomokiyo A, Wada N, Maeda H.Periodontal Ligament Stem Cells: Regenerative Potency in Periodontium[J].Stem Cells Dev, 2019, 28(15):974-985
[3]	Oshima M, Tsuji T.Whole Tooth Regeneration as a Future Dental Treatment[J].Adv Exp Med Biol, 2015, 881:255-269
[4]	梁莉，余继锋，周威，等.内皮素-对牙周膜干细胞成骨分化能力的影响[J].解放军医学院学报, 2016, 37(9):988-991
[5]	Liang L, Zhou W, Yang N, et al.ET-1 promotes differentiation of periodontal ligament stem cells into osteoblaststhrough ETR,MAPK and Wntβ-catenin signaling pathways under inflammatory microenvironment[J].Mediators Inflamm., 2016, 2016:8467849-
[6]	Yanagisawa M, Kurihara H, Kimura S, et al.A novel potent vasoconstrictor peptide produced by vascular endothelial cells[J].Nature, 1988, 332(6163):411-415
[7]	Levin E R.Endothelins[J].N Engl J Med, 1995, 333(6):356-363
[8]	Clines GA, Mohammad KS, Grunda JM et al.Regulation of Postnatal Trabecular Bone Formation by the Osteoblast Endothelin A Receptor[J].J Bone Miner Res, 2011, 26（10）:2523-2536
[9]	Lee MS, Wang J, Yuan H et al.Endothelin-1 differentially directs lineage specification of adipose- and bone marrow-derived mesenchymal stem cells[J].FASEB J, 2019, 33(1):996-1007
[10]	Tsukahara H, Sekine K, Miura M, et al.Vasoactive and natriuretic mediators in umbilical cord blood: a report of our observation and review of the literature[J].Early Hum Dev, 2002, 69(1-2):57-64
[11]	Yuan W, Li ZX, Zhao CL, et al.Attenuation of the degenerative effects of endothelin-1 on cartilaginous end plate cells by the endothelin receptor antagonist BQ-123 via the Wntβ-catenin signaling pathway[J].Spine J, 2018, 18(9):1669-1677
[12]	Hu LW, Wang X, Jiang XQ, et al.In vivo and in vitro study of osteogenic potency of endothelin-1 on bone marrow-derived mesenchymal stem cells[J].Exp Cell Res, 2017, 357(1):25-32
[13]	Han H.RNA Interference to Knock Down Gene Expression[J].Methods in molecular biology, 2018, 1706:293-302
[14]	Bodak M, Cirera-Salinas D, Luitz J, et al.The Role of RNA Interference in Stem Cell Biology: Beyond the Mutant Phenotypes[J].J Mol Biol, 2017, 429(10):1532-1543

[1]	范瑞瑞李欣聪刘玉孙卫斌郭婷. 聚己内酯/Ⅰ型胶原/氟磷灰石复合支架的制备及生物相容性研究[J]. , 2019, 10(1): 12-16.
[2]	石安源鲍东昱童昕秦海燕王斌. SB431542对人牙龈间充质干细胞体外成骨分化的影响[J]. , 2019, 10(1): 6-11.
[3]	杨昊清范志朋. SFRP2对脂肪干细胞成骨分化的作用研究[J]. , 2018, 9(3): 120-124.
[4]	王红魏福兰王春玲. miR-21研究进展[J]. , 2018, 9(2): 106-109.
[5]	周五超甘烟紫习伟宏. 多孔TiO2修饰钛植体对骨髓间充质干细胞粘附和成骨分化的影响[J]. , 2018, 9(2): 74-78.
[6]	王海锋幸丹刘亚男刘思思宁美芝李文峰曹钰. 过表达BCOR-short基因抑制根尖牙乳头干细胞成骨分化功能[J]. , 2018, 9(1): 12-16.
[7]	张文方明鲍东昱丁晓晨秦海燕童昕. DAPT对牙周膜干细胞体外增殖及成骨分化的影响[J]. , 2018, 9(1): 7-11.
[8]	梁晓伟李阳飞李琥陈文静. 羟基红花黄色素A对人骨髓间充质干细胞成骨分化的影响[J]. , 2017, 8(2): 90-94.
[9]	傅昱瑾曹灵俞艳金琳徐云龙汪延秋于金华. let-7c对大鼠骨髓间充质干细胞增殖及成牙/成骨分化能力的影响[J]. , 2017, 8(2): 65-74.
[10]	谭学莲黄定明张岚. 叉头框蛋白O转录因子调节骨平衡[J]. , 2016, 7(1): 42-45.
[11]	陈小燕丁寅金岩. TNF-α通过NF-κB信号通路对人牙周膜干细胞成骨分化调控作用的研究[J]. , 2015, 6(3): 124-128.
[12]	孔祥伟叶瑞东刘文佳丁寅金岩. 炎症微环境中经典Wnt 信号通路对牙周膜干细胞成骨分化的调控作用[J]. , 2015, 6(3): 129-136.
[13]	袁林孙晋杨征毅曹依娜潘广嗣钱钧何恩亮王涵. 乙酰基转移酶KAT2A调控牙周膜干细胞成骨分化的研究[J]. , 2015, 6(3): 155-159.
[14]	束丽红曹灵闫明李俊俊张光东于金华. 不同发育阶段的人牙周膜干细胞增殖能力和成牙/成骨能力的比较研究[J]. , 2013, 4(2): 65-69.
[15]	徐炜王明国马锋杨世茂林夏莲于彦领张新新. 富血小板纤维蛋白对兔脂肪干细胞成骨分化的影响[J]. , 2013, 4(1): 19-22.