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高糖对牙髓干细胞的影响、信号通路及应用研究

The investigation of effects, signal pathways, and applications of high glucose on dental pulp stem cells.

作者信息

Lee Shih-Yu, Li I-Hsun, Tsai Wei-Cheng, Ho Ming-Hua, Li Chung-Hsing

机构信息

Graduate Institute of Aerospace and Undersea Medicine, National Defense Medical Center, Taipei, Taiwan.

School of Pharmacy, National Defense Medical Center, Taipei, Taiwan.

出版信息

J Dent Sci. 2025 Jul;20(3):1731-1738. doi: 10.1016/j.jds.2025.03.038. Epub 2025 Apr 12.

Abstract

BACKGROUND/PURPOSE: Dental pulp stem cells (DPSCs) are among the most widely used dental-derived mesenchymal stem cells (MSCs), and their applications have involved various regions. The glucose metabolism plays a key role in cell function and the current literature presents conflicting evidence regarding the influence of glucose on MSCs' properties. This study evaluated the impact of high glucose (HG) on DPSCs.

MATERIALS AND METHODS

DPSCs were stimulated with indicated concentrations of glucose. Cell viability was assessed using a cell counting kit, while apoptosis and autophagy were analyzed via western blot. MSCs immunophenotypic properties were determined by flow cytometry. Osteogenic, adipogenic, and neurogenic differentiation potential were evaluated using western blot, Alizarin red staining, oil red-O staining, and morphological analysis.

RESULTS

HG exposure led to a significant decrease in cell viability, with increased apoptosis and autophagy, as indicated by increased levels of cleaved caspase-3, cleaved poly (ADP-ribose) polymerase (PARP), and an elevated microtubule-associated protein 1 light chain 3 beta (LC3B)-II/LC3B-I ratio. However, the immunophenotypic characteristics of DPSCs remained unchanged. DPSCs also demonstrated enhanced osteogenic, adipogenic, and neurogenic differentiation potential by expressing Alizarin red and oil red-O staining, neural-like cell morphology, and several differentiation-related proteins after HG culture stimulation.

CONCLUSION

The present study demonstrated that while HG slightly impairs DPSC viability, it promotes osteogenic, adipogenic, and neurogenic differentiation. Providing valuable insights into the mechanisms by which HG influences various differentiation pathways in DPSCs and establishes a foundation for potential clinical applications of DPSCs in regenerative medicine for diabetic patients.

摘要

背景/目的:牙髓干细胞(DPSCs)是应用最为广泛的牙源性间充质干细胞(MSCs)之一,其应用涉及多个领域。葡萄糖代谢在细胞功能中起关键作用,而目前的文献对于葡萄糖对MSCs特性的影响存在相互矛盾的证据。本研究评估了高糖(HG)对DPSCs的影响。

材料与方法

用指定浓度的葡萄糖刺激DPSCs。使用细胞计数试剂盒评估细胞活力,通过蛋白质免疫印迹法分析细胞凋亡和自噬。通过流式细胞术测定MSCs的免疫表型特性。使用蛋白质免疫印迹法、茜素红染色、油红O染色和形态学分析评估成骨、成脂和神经分化潜能。

结果

HG暴露导致细胞活力显著降低,细胞凋亡和自噬增加,表现为裂解的半胱天冬酶-3、裂解的聚(ADP-核糖)聚合酶(PARP)水平升高以及微管相关蛋白1轻链3β(LC3B)-II/LC3B-I比值升高。然而,DPSCs的免疫表型特征保持不变。在HG培养刺激后,DPSCs通过茜素红和油红O染色、神经样细胞形态以及几种分化相关蛋白的表达,还表现出增强的成骨、成脂和神经分化潜能。

结论

本研究表明,虽然HG会轻微损害DPSC活力,但它能促进成骨、成脂和神经分化。这为HG影响DPSCs中各种分化途径的机制提供了有价值的见解,并为DPSCs在糖尿病患者再生医学中的潜在临床应用奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/986c/12254765/4e80ea27662c/gr1.jpg

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