High glucose induces a metabolic memory in human periodontal ligament cells

doi:10.3969/j.issn.2095-4344.2017.04.007

Abstract

Abstract:

Abstract
BACKGROUND: Studies on high glucose exposure in human periodontal ligament cells usually focus on the biological behaviors, pathways and secretory factors, but whether the metabolic memory is involved is little known.
OBJECTIVE: To investigate the metabolic memory of high glucose exposure in human periodontal ligament cells.
METHODS: Human periodontal ligament cells were primarily cultured and identified. Cells at 5-8 passages were selected and randomized into four groups. Group A (controls): DMEM containing 5.5 mmol/L glucose for 8 days; group B (5-day memory group): DMEM containing 35 mmol/L glucose for 3 days and DMEM containing 5.5 mmol/L glucose for 5 days; group C (3-day memory group): DMEM containing 35 mmol/L glucose for 5 days and DMEM containing 5.5 mmol/L glucose for 3 days; group D (8-day high glucose group): DMEM containing 35 mmol/L glucose for 8 days. The cell proliferation was detected by cell counting kit-8, the cell apoptosis was determined by flow cytometry, and the levels of total proteins and alkaline phosphatase were investigated using ELISA.
RESULTS AND CONCLUSION: Compared with the control group, the cell proliferation in the other three groups was significantly reduced (P < 0.05), the number of apoptotic cells was significantly increased, while the levels of total proteins and alkaline phosphatase were significantly decreased (P < 0.05). These results suggest that high glucose causes persistent changes in human periodontal ligament cells by inhibiting cell viability, increasing the apoptosis and downregulating the levels of the total proteins and alkaline phosphatase

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Tissue Engineering, Apoptosis, Alkaline Phosphatase

CLC Number:

R318

Ren Wei-wei1, Li Shou-hong2, Xiong Jie1, Zhang Fan1, Guan Qin1. High glucose induces a metabolic memory in human periodontal ligament cells[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(4): 530-537.

Figures/Tables 2

2.1 人牙周膜细胞的形态及组织来源鉴定结果培养4- 7 d后即可以见到人牙周膜细胞从组织块周围游出。细胞伸展，形态为星形或长梭形。相互交织成网状。在无组织块的空白区域可见散在零星细胞。随着细胞密度增加，细胞形态为细长梭形，以组织块为中心排列成放射状。免疫组织化学染色显示抗波形丝蛋白多克隆抗体表达阳性，胞浆呈现棕黄色，抗角蛋白表达阴性(图1)。 2.2 各组人牙周膜细胞增殖的检测结果 35 mmol/L葡萄糖对人牙周膜细胞增殖有抑制作用。高糖刺激3 d转换为5.5 mmol/L正常葡萄糖培养5 d后人牙周膜细胞活力相比全程高糖组有所恢复(P < 0.05)，但与正常糖比较细胞活力仍有降低(P < 0.05)。35 mmol/L高糖刺激人牙周膜细胞5 d后用5.5 mmol/L正常葡萄糖浓度进行培养，时间为3 d，相比于正常葡萄糖组，细胞活力显著下降(P < 0.05)，与高糖刺激3 d组比较活力仍有降低(P < 0.05)(图2)。 2.3 高糖对人牙周膜细胞凋亡的影响 35 mmol/L葡萄糖对人牙周膜细胞凋亡有明显增强作用。高糖刺激3 d转换为5.5 mmol/L正常葡萄糖培养5 d后人牙周膜细胞凋亡细胞相比全程高糖组有所减少(P < 0.05)，但与正常糖比较仍有增加(P < 0.05)。35 mmol/L高糖刺激人牙周膜细胞 5 d后用5.5 mmol/L正常葡萄糖浓度进行培养，时间为3 d，相比于正常葡萄糖组，凋亡细胞显著增加(P < 0.05)，与高糖刺激3 d组比较凋亡细胞仍有增强(P < 0.05)(图3，4)。 2.4 高糖对人牙周膜细胞碱性磷酸酶活性的影响人牙周膜细胞在35 mmol/L时的碱性磷酸酶活性比5.5 mmol/L葡萄糖时明显降低抑制(P < 0.05)。高糖刺激3 d转换为 5.5 mmol/L正常葡萄糖培养5 d后人牙周膜细胞碱性磷酸酶活性相比全程高糖组有所回升(P < 0.05)，但与正常糖比较仍有差距(P < 0.05)。35 mmol/L高糖刺激人牙周膜细胞5 d转换为5.5 mmol/L正常葡萄糖培养3 d后，人牙周膜细胞碱性磷酸酶活性抑制情况相比相比全程高糖组无明显缓解(P < 0.05)(图5)。"

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