儿童脱落乳牙牙髓干细胞（SHED）的再生活性受 TGF-β1 的调控，这种调控与 ALK5/Smad2、TAK1、p38 和 MEK/ERK 信号通路有关。
Regulation of the regenerative activity of dental pulp stem cells from exfoliated deciduous teeth (SHED) of children by TGF-β1 is associated with ALK5/Smad2, TAK1, p38 and MEK/ERK signaling.
机构信息
Department of Dentistry, National Taiwan University Hospital, and School of Dentistry, National Taiwan University Medical College, Taipei, Taiwan.
Chang Gung University of Science and Technology, Kwei-Shan, Taoyuan, Taiwan.
出版信息
Aging (Albany NY). 2020 Nov 4;12(21):21253-21272. doi: 10.18632/aging.103848.
Transforming growth factor-β1 (TGF-β1) regulates wound healing/regeneration and aging processes. Dental pulp stem cells from human exfoliated deciduous teeth (SHED) are cell sources for treatment of age-related disorders. We studied the effect of TGF-β1 on SHED and related signaling. SHED were treated with TGF-β1 with/without pretreatment/co-incubation by SB431542, U0126, 5Z-7-oxozeaenol or SB203580. Sircol collagen assay, 3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) assay, RT-PCR, western blotting and PathScan phospho-ELISA were used to measure the effects. We found that SHED expressed ALK1, ALK3, ALK5, TGF-RII, betaglycan and endoglin mRNA. TGF-β1 stimulated p-Smad2, p-TAK1, p-ERK, p-p38 and cyclooxygenase-2 (COX-2) protein expression. It enhanced proliferation and collagen content of SHED that were attenuated by SB431542, 5Z-7-oxozeaenol and SB203580, but not U0126. TGF-β1 (0.5-1 ng/ml) stimulated ALP of SHED, whereas 5-10 ng/ml TGF-β1 suppressed ALP. SB431542 reversed the effects of TGF-β1. However, 5Z-7-oxozeaenol, SB203580 and U0126 only reversed the stimulatory effect of TGF-β1 on ALP. Four inhibitors attenuated TGF-β1-induced COX-2 expression. TGF-β1-stimulated TIMP-1 and N-cadherin was inhibited by SB431542 and 5Z-7-oxozeaenol. These results indicate that TGF-β1 affects SHED by differential regulation of ALK5/Smad2/3, TAK1, p38 and MEK/ERK. TGF-β1 and SHED could potentially be used for tissue engineering/regeneration and treatment of age-related diseases.
转化生长因子-β1（TGF-β1）调节伤口愈合/再生和衰老过程。人乳牙脱落牙髓干细胞（SHED）是治疗与年龄相关疾病的细胞来源。我们研究了 TGF-β1 对 SHED 及相关信号的影响。用 TGF-β1 处理 SHED，并用 SB431542、U0126、5Z-7-氧杂豆甾醇或 SB203580 预处理/共孵育。用 Sircol 胶原测定法、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐（MTT）测定法、碱性磷酸酶（ALP）测定法、RT-PCR、western blot 和 PathScan 磷酸化 ELISA 来测量效果。我们发现 SHED 表达 ALK1、ALK3、ALK5、TGF-RII、β糖蛋白和内格林 mRNA。TGF-β1 刺激 p-Smad2、p-TAK1、p-ERK、p-p38 和环氧化酶-2（COX-2）蛋白表达。它增强了 SHED 的增殖和胶原含量，而 SB431542、5Z-7-氧杂豆甾醇和 SB203580 则减弱了这一作用，但 U0126 则没有。TGF-β1（0.5-1ng/ml）刺激 SHED 的 ALP，而 5-10ng/ml TGF-β1 则抑制 ALP。SB431542 逆转了 TGF-β1 的作用。然而，5Z-7-氧杂豆甾醇、SB203580 和 U0126 仅逆转了 TGF-β1 对 ALP 的刺激作用。四种抑制剂减弱了 TGF-β1 诱导的 COX-2 表达。TGF-β1 刺激 TIMP-1 和 N-钙粘蛋白的表达被 SB431542 和 5Z-7-氧杂豆甾醇抑制。这些结果表明，TGF-β1 通过对 ALK5/Smad2/3、TAK1、p38 和 MEK/ERK 的不同调节来影响 SHED。TGF-β1 和 SHED 可能被用于组织工程/再生和治疗与年龄相关的疾病。
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