中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (13): 1992-1997.doi: 10.3969/j.issn.2095-4344.2017.13.005

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

维生素C促进高糖环境下人脂肪干细胞的增殖及相关机制

李江锋，丁世超，戚亚伟，李  瑾，曾国芳，赖  巧，刘  莉，张培华   

  1. 广东医科大学附属医院整形外科研究所，广东省湛江市  524001
  • 修回日期:2017-02-12 出版日期:2017-05-08 发布日期:2017-06-09
  • 通讯作者: 张培华，博士，主任医师，硕士生导师，广东医科大学附属医院整形外科研究所，湛江市 524001；刘莉，博士，助理研究员，广东医科大学附属医院整形外科研究所，广东省湛江市 524001
  • 作者简介:李江锋，男，1991年生，广东省陆丰市人，汉族，广东医科大学在读硕士，主要从事脂肪干细胞的基础与临床研究。
  • 基金资助:

    2015年国家自然科学基金面上项目(81570260)；2014年广东省自然科学基金(2014A030313535)；2016广东省科技计划项目(2016A020214018)；2014湛江市财政资金科技专项经费资助项目(2014C01022)；2015广东医科大学博士基金(BJ201510)；2015年广东医科大学附属医院博士启动基金(BJ201505)

Vitamin C treatment promotes cell proliferation of human adipose-derived stem cells under high glucose conditions

Li Jiang-feng, Ding Shi-chao, Qi Ya-wei, Li Jin, Zeng Guo-fang, Lai Qiao, Liu Li, Zhang Pei-hua   

  1. Institute of Plastic Surgery, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, Guangdong Province, China
  • Revised:2017-02-12 Online:2017-05-08 Published:2017-06-09
  • Contact: Zhang Pei-hua, M.D., Chief physician, Master’s supervisor, Institute of Plastic Surgery, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, Guangdong Province, China；Liu Li, M.D., Assistant researcher, Institute of Plastic Surgery, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, Guangdong Province, China
  • About author:Li Jiang-feng, Studying for master’s degree, Institute of Plastic Surgery, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, Guangdong Province, China
  • Supported by:

    the National Natural Science Foundation of China in 2015, No. 81570260; the Natural Science Foundation of Guangdong Province in 2014, No. 2014A030313535; the Science and Technology Plan of Guangdong Province in 2016, No. 2016A020214018; the Special Financial Funds for Science and Technology in Zhanjiang in 2014, No. 2014C01022; the Doctor Fund of Guangdong Medical University in 2015, No. BJ201510; the Doctoral Initial Fund of the Affiliated Hospital of Guangdong Medical University in 2015, No. BJ201505

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291

被引次数

4

摘要：

文章快速阅读：

文题释义：
干细胞治疗糖尿病溃疡创面：间充质干细胞不仅可分化为组织修复所需要的内皮细胞，还可分泌各种细胞因子。来源方便且获取容易的脂肪干细胞移植可明显改善伤口愈合，研究表明，将种植在脱细胞真皮基质的脂肪干细胞移植到切除伤口模型糖尿病大鼠，明显提高了毛细血管密度及血管新生，加速了伤口愈合。移植骨髓来源的间充质干细胞能通过促进组织再生，显著改善糖尿病大鼠背侧皮肤缺损的恢复。
维生素C的功效：维生素C除了具有抗氧化作用外，还具有促进细胞增殖的重要作用，对干细胞的生长、增殖和分化都有影响；据报道，在培养基中加入合适剂量的维生素C，能够增加体细胞中诱导多能干细胞的产生。但是至今为止，维生素C对高糖环境下人脂肪干细胞的增殖作用及相关分子机制目前尚缺乏研究。

 

摘要
背景：研究证实，维生素C除了具有抗氧化作用外，还具有促进细胞增殖的重要作用，但至今为止，有关维生素C对高糖环境下人脂肪干细胞的增殖作用及相关分子机制尚不明确。
目的：研究维生素C促进高糖环境下人脂肪干细胞增殖的作用及相关分子机制。
方法：取高塘环境中培养的第3代人脂肪干细胞，分别以0，100，150，200，250，300 μmol/L维生素C处理，以低糖环境中培养的细胞为对照，MTT法筛选最佳维生素C干预浓度及处理时间，进行Western Blot检测。EdU检测：将高糖环境中培养的人脂肪干细胞分4组培养，空白对照组不进行任何干预，维生素C组加入最佳浓度维生素C，LY294002组加入PI3K/AKT信号通路抑制剂LY294002与最佳浓度维生素C，U0126组加入ERK/MAPK信号通路抑制剂U0126与最佳浓度维生素C，以低糖环境中培养的细胞为对照，干预48 h后，检测EdU标记阳性细胞数。
结果与结论：①CCK-8法检测：200 μmol/L维生素C促进高糖环境下人脂肪干细胞的增殖作用最强，所以此为最佳干预浓度，最佳处理时间为48 h；②Western Blot检测：对照组、0 μmol/L维生素C处理组、200 μmol/L维生素C处理组p-AKT蛋白表达无差异，200 μmol/L维生素C处理组p-ERK蛋白表达高于0 μmol/L维生素C处理组(P < 0.01)；③EdU检测：维生素C组、LY294002组、对照组EdU标记阳性细胞数高于空白对照组(P < 0.01)，维生素C组EdU标记阳性细胞数高于U0126组(P < 0.01)；④结果表明：维生素C通过ERK/MAPK信号通路促进高糖环境下人脂肪干细胞的增殖。

 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-8951-0992(李江锋)

关键词: 干细胞, 脂肪干细胞, 维生素C, 糖尿病难愈创面, 高糖环境, 增殖, ERK/MAPK信号通路, 国家自然科学基金

Abstract:

BACKGROUND: Although vitamin C has an anti-oxidation role and can promote cell proliferation, there is a lack of research about the promoting effect of vitamin C on the proliferation of adipose-derived stem cells under high glucose conditions and the related molecular mechanisms.
OBJECTIVE: To explore the promoting effect of vitamin C on the proliferation adipose-derived stem cells treated by the high glucose and the related molecular mechanisms. 
METHODS: Passage 3 human adipose-derived stem cells were cultured under high glucose conditions and then treated with different concentrations of vitamin C (0, 100, 150, 200, 250, 300 μmol/L). Cells cultured under low glucose conditions acted as controls. The expression levels of p-ERK and p-AKT proteins were detected by western blot. MTT method was used to choose the optimal concentration and time of vitamin C for all the subsequent tests. Human adipose-derived stem cells cultured under high glucose conditions were divided into four groups, and cells in blank control group had no treatment. Cells in the other three groups were treated with the optimal concentration of vitamin C (vitamin C group), LY294002+the optimal concentration of vitamin C (LY294002 group), or U0126+the optimal concentration of vitamin C (U0126 group) for 48 hours. EdU staining assay was used to detect the cell proliferation of human adipose-derived stem cells. 
RESULTS AND CONCLUSION: (1) Cell counting kit detection: We found that high glucose reduced the proliferation of human adipose-derived stem cells, and vitamin C promoted the proliferation of these cells. The best concentration of vitamin C was 200 μmol/L and the optimal effect time was 48 hours. (2) Western blot detection: Compared with the 0 μmol/L vitamin C group, the level of p-ERK in the 200 μmol/L vitamin C group was upregulated significantly (P < 0.01), while no significant expression change in p-AKT protein was found in control, 0 and 200 μmol/L vitamin C groups. (3) EdU test: the number of EdU positive cells was significantly higher in the vitamin C, LY294002, and control groups compared with the blank control group (P < 0.01). Moreover, compared with the vitamin C group, the EdU positive cells in the U0126 group were decreased significantly in number (P < 0.01). In conclusion, the ERK/MAPK signaling pathway is involved in the promotion effect of vitamin C on the proliferation of human adipose-derived stem cells under high glucose conditions.

 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Ascorbic Acid, Cell Proliferation, Tissue Engineering

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