Asghari Mona, Nasoohi Nikoo, Hodjat Mahshid
Department of Biochemistry and Biophysics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, Iran.
Dent Med Probl. 2021 Jan-Mar;58(1):39-46. doi: 10.17219/dmp/130090.
Diabetes is one of the most common metabolic diseases that disrupt the functioning of different body organs, including oral tissue. Some diabetic complications remain even after the control of the hyperglycemic condition. The adverse effect of hyperglycemia on the pulp structure and function has been shown previously.
The purpose of this study was to investigate the effect of the hyperglycemic state on the aging of pulp cells and evaluate the role of Wnt signaling as the underlying mechanism of this process.
The isolated pulp cells were cultured in the Minimum Essential Medium (MEM)-alpha for 7, 14 and 21 days under the influence of glucose at concentrations of 5 mM, 20 mM and 30 mM. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate cell viability, the beta-galactosidase test was applied to assess cellular senescence and gene expression was measured with quantitative real-time polymerase chain reaction (qRT-PCR).
The results of this study showed that cell proliferation decreased following exposure to 20 and 30 mM glucose, which was accompanied by an increased number of senescent cells and an increased p21 expression. There was a significant increase in beta-catenin and Wnt1 expression in response to high glucose. Treatment with beta-catenin inducer enhanced cellular aging in the hyperglycemic state, while betacatenin inhibitor decreased the senescence response.
The present study further confirmed the effect of the high-glucose condition on pulp cell aging and suggests a role for beta-catenin in the induction of cellular aging. Targeting the key regulators of cellular aging in pulp tissue might lead to the development of new therapies for the prevention and treatment of endodontic complications in diabetic patients.
糖尿病是最常见的代谢性疾病之一，会干扰包括口腔组织在内的不同身体器官的功能。即使在高血糖状况得到控制后，一些糖尿病并发症仍然存在。高血糖对牙髓结构和功能的不良影响此前已有报道。
本研究旨在探讨高血糖状态对牙髓细胞衰老的影响，并评估Wnt信号通路作为该过程潜在机制的作用。
将分离的牙髓细胞在含有5 mM、20 mM和30 mM葡萄糖的影响下，于最低基本培养基（MEM）-α中培养7天（天）、14天和21天。采用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）法评估细胞活力，应用β-半乳糖苷酶试验评估细胞衰老，并通过定量实时聚合酶链反应（qRT-PCR）测量基因表达。
本研究结果表明，暴露于20 mM和30 mM葡萄糖后细胞增殖减少，同时衰老细胞数量增加且p21表达增加。高糖刺激后β-连环蛋白和Wnt1表达显著增加。用β-连环蛋白诱导剂处理可增强高血糖状态下的细胞衰老，而β-连环蛋白抑制剂可降低衰老反应。
本研究进一步证实了高糖状况对牙髓细胞衰老的影响，并提示β-连环蛋白在诱导细胞衰老中起作用。针对牙髓组织中细胞衰老的关键调节因子可能会开发出预防和治疗糖尿病患者牙髓并发症的新疗法。
