褪黑素对人牙周膜细胞成骨分化能力的影响

摘要/Abstract

摘要： 目的探讨褪黑素对人牙周膜细胞（hPDLCs）成骨分化能力的影响。方法原代培养法成功提取hPDLCs后，加入不同浓度的褪黑素作为实验组，空白对照组不加褪黑素。MTT法检测不同浓度褪黑素对细胞增殖的影响，相关试剂盒检测碱性磷酸酶（ALP）的活性和表达后，采用茜素红S染色、Real-time PCR和Western Blot分别研究褪黑素对hPDLCs的矿化结节、成骨相关mRNA以及蛋白表达的影响。结果与对照组相比，褪黑素组hPDLCs的增殖活力略有抑制，其中50 μmol/L和100 μmol/L褪黑素组抑制最为显著（P＜0.01）；褪黑素组均表现为ALP活性和表达上升，矿化结节形成增多且呈浓度依赖性（P＜0.01）；Real-time PCR和Western Blot结果显示褪黑素组成骨相关mRNA和蛋白表达均有上升（P＜0.01）。结论褪黑素可促进hPDLCs的成骨分化，具有作为牙周再生治疗药物的潜能。

关键词: 褪黑素, 牙周炎, 成骨分化, 人牙周膜细胞

Abstract: Objective　 To evaluate the effects of melatonin on the function of osteogenic differentiation of human periodontal ligament cells (hPDLCs).　Methods　 Melatonin was added into hPDLCs as experimental group. The blank control group was not added with melatonin. The effect of different concentrations of melatonin on proliferation of hPDLCs was assessed through MTT assay. Alkaline phosphatase (ALP) activity assay, ALP staining, Alizarin red S staining, Real-time PCR and Western Blot were used to observe the ALP activity, formation of mineral nodules, mRNA and protein expression of hPDLCs, respectively.　Results　 Compared to the control group, the proliferation of hPDLCs in the melatonin groups decreased. Melatonin was shown to enhance the ALP activity and expression of hPDLCs. The melatonin groups were observed more mineral nodules compared to the control group. Real-time PCR and Western Blot results showed that the mRNA and protein expression of Col-I, Runx2 and OSX increased by melatonin in different concentration.　Conclusions　 Melatonin has a promoting effect on the osteogenic differentiation of hPDLCs, and it has further research on the value of periodontal regenerative therapy.

Key words: Melatonin, Periodontitis, Osteogenesis, Human periodontal ligament cells

中图分类号:

R349.5

张梦涵何智琪邓辉胡荣党. 褪黑素对人牙周膜细胞成骨分化能力的影响[J]. 口腔医学, 2021, 41(3): 198-203.

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