人牙周膜细胞与高糖损伤的代谢记忆效应  

High glucose induces a metabolic memory in human periodontal ligament cells

作　　者：任伟伟[1] 李守宏[2] 熊洁[1] 张帆[1] 管琴[1] Ren Wei-wei Li Shou-hong Xiong Jie Zhang Fan Guan Qin(Department of Prevention Dentistry, Dongfeng Stomatological Hospital, Hubei University of Medicine, Shiyan 442001, Hubei Province, China Department of Oral and Maxillofacial Surgery, Dongfeng Stomatological Hospital, Hubei University of Medicine, Shiyan 442001, Hubei Province, China)

机构地区：[1]湖北医药学院附属东风口腔医院预防保健科,湖北省十堰市442001 [2]湖北医药学院附属东风口腔医院颌面外科,湖北省十堰市442001

出　　处：《中国组织工程研究》2017年第4期532-537,共6页Chinese Journal of Tissue Engineering Research

基　　金：湖北省教育厅青年人才项目(Q20162108);十堰市科技局计划项目(15y50)~~

摘　　要：背景:目前高糖影响人牙周膜细胞相关研究多集中在细胞生物学行为、相关通路、因子分泌研究等方面,但对于人牙周膜细胞对高糖影响是否存在"代谢记忆"效应的报道较少。目的:观察人牙周膜细胞对高糖损伤是否存在记忆效应。方法:原代培养并鉴定人牙周膜细胞,实验采用5-8代细胞,划分为对照组、5 d记忆组、3 d记忆组、8 d高糖组:对照组葡萄糖浓度为5.5 mmol/L DMEM培养液培养8 d;5 d记忆组用35 mmol/L DMEM培养液刺激3 d后5.5 mmol/L DMEM培养液培养5 d;3 d记忆组35 mmol/L DMEM培养液刺激5 d后5.5 mmol/L DMEM培养液培养3 d;8 d高糖组葡萄糖浓度35 mmol/L DMEM培养液进行8 d的刺激。细胞增殖通过CCK-8法进行测定,细胞凋亡通过流式细胞仪进行检测,采用酶标仪检测总蛋白的合成及碱性磷酸酶水平变化。结果与结论:与对照组比较,其他3组中的细胞增殖速度明显降低(P<0.05)。细胞凋亡明显升高,总蛋白合成以及碱性磷酸酶分泌水平,所有高糖组总蛋白合成以及碱性磷酸酶分泌水平则明显下降(P<0.05)。结果说明,人牙周膜细胞在高糖环境中产生的损伤具有记忆效应,以细胞活力长时间下降、凋亡增加、总蛋白合成、碱性磷酸酶水平持续下调为主要表现。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; groupB（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

关 键 词：组织工程 细胞凋亡 碱性磷酸酶 组织构建 高糖 人牙周膜细胞 代谢记忆 增殖 凋亡 流式细胞术 总蛋白合成 细胞生物学活性 

分 类 号：R318[医药卫生—生物医学工程]