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胰岛素样生长因子-1 在高糖条件下促进人牙髓细胞的增殖和成牙本质细胞分化。

Insulin-like growth factor-1 promotes the proliferation and odontoblastic differentiation of human dental pulp cells under high glucose conditions.

机构信息

Department of Endodontics, School of Stomatology, China Medical University, Shenyang, Liaoning 110002, P.R. China.

Department of Periodontics, School of Stomatology, China Medical University, Shenyang, Liaoning 110002, P.R. China.

出版信息

Int J Mol Med. 2017 Oct;40(4):1253-1260. doi: 10.3892/ijmm.2017.3117. Epub 2017 Sep 1.

Abstract

Insulin-like growth factor-1 (IGF-1) promotes human dental pulp stem cell proliferation and osteogenic differentiation. However, the effects of IGF-1 on the proliferation, apoptosis and odontoblastic differentiation (mineralization) of dental pulp cells (DPCs) under high glucose (GLU) conditions remain unclear. In this study, isolated primary human DPCs were treated with various concentrations of high GLU. Cell proliferation and apoptosis were determined by Cell Counting Kit-8 and Annexin V-FITC/PI assays, respectively. The cells were cultured in odontoblastic induction medium containing various concentrations of high GLU. Odontoblastic differentiation was determined by alkaline phosphatase (ALP) activity assay. Mineralization formation was evaluated by von Kossa staining. The expression levels of IGF family members were measured by western blot analysis and RT-qPCR during proliferation and differentiation. The cells were then exposed to 25 mM GLU and various concentrations of IGF-1. Cell proliferation, apoptosis, ALP activity, mineralization formation and the levels of mineralization-related proteins were then evaluated. Our results revealed that high GLU significantly inhibited cell proliferation and promoted cell apoptosis. GLU (25 and 50 mM) markedly reduced ALP activity and mineralization on days 7 and 14 after differentiation. The levels of IGF family members were markedly decreased by high GLU during proliferation and differentiation. However, IGF-1 significantly reversed the effects of high GLU on cell proliferation and apoptosis. Additionally, IGF-1 markedly restored the reduction of ALP activity and mineralization induced by high GLU. Our findings thus indicate that IGF-1 attenuates the high GLU-induced inhibition of DPC proliferation, differentiation and mineralization.

摘要

胰岛素样生长因子-1(IGF-1)可促进人牙髓干细胞增殖和成骨分化。然而,IGF-1 对高糖(GLU)条件下牙髓细胞(DPCs)增殖、凋亡和成牙本质分化(矿化)的影响尚不清楚。在本研究中,用不同浓度的高 GLU 处理分离的原代人牙髓细胞。通过 Cell Counting Kit-8 和 Annexin V-FITC/PI 检测分别测定细胞增殖和凋亡。将细胞在含有不同浓度高 GLU 的成牙本质诱导培养基中培养。碱性磷酸酶(ALP)活性测定法测定成牙本质分化。von Kossa 染色评价矿化形成。在增殖和分化过程中,通过 Western blot 分析和 RT-qPCR 测定 IGF 家族成员的表达水平。然后将细胞暴露于 25 mM GLU 和不同浓度的 IGF-1 中。然后评估细胞增殖、凋亡、ALP 活性、矿化形成和矿化相关蛋白的水平。我们的结果表明,高 GLU 显著抑制细胞增殖并促进细胞凋亡。GLU(25 和 50 mM)在分化后第 7 天和第 14 天显著降低了 ALP 活性和矿化。在增殖和分化过程中,高 GLU 显著降低了 IGF 家族成员的水平。然而,IGF-1 显著逆转了高 GLU 对细胞增殖和凋亡的影响。此外,IGF-1 显著恢复了高 GLU 诱导的 ALP 活性和矿化减少。我们的研究结果表明,IGF-1 可减轻高 GLU 诱导的 DPC 增殖、分化和矿化抑制。

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