中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (29): 4701-4706.doi: 10.3969/j.issn.2095-4344.1798

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

罗汉果皂苷V刺激成骨细胞增殖与分化

姚顺晗，廖  亮，覃家港，韦华成   

  1. 广西医科大学第一附属医院，广西壮族自治区南宁市  530021
  • 修回日期:2019-05-07 出版日期:2019-10-18 发布日期:2019-10-18
  • 通讯作者: 廖亮，硕士，副主任医师，广西医科大学第一附属医院，广西壮族自治区南宁市 530021
  • 作者简介:姚顺晗，男，1993年生，广西壮族自治区田林县人，汉族，广西医科大学在读硕士，主要从事骨质疏松症的预防与治疗研究。
  • 基金资助:

    广西自然科学基金青年基金一般项目(2016GXNSFBA380103)，项目负责人：廖亮

Mogroside V promotes bone formation by stimulating osteoblast proliferation and differentiation

Yao Shunhan, Liao Liang, Qin Jiagang, Wei Huacheng   

  1. First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Revised:2019-05-07 Online:2019-10-18 Published:2019-10-18
  • Contact: Liao Liang, Master, Associate chief physician, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • About author:Yao Shunhan, Master candidate, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Supported by:

    the Natural Science Foundation of Guangxi Zhuang Autonomous Region (Youth Project), No. 2016GXNSFBA380103 (to LL)

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摘要：

文章快速阅读：

文题释义：
罗汉果皂苷V：是罗汉果中主要化学成分之一，其具有甜度高、热量低、水溶性好等特点，可以用作甜味剂，尤其适合肥胖和高血糖患者。传统中医认为其具有清热润肺、利咽开音、滑肠通便的功效，而近年有研究表明其具有抗氧化、降血糖、抗癌等多种药效，但尚未见报道其对成骨细胞的作用。
成骨细胞：由间充质干细胞分化形成，其不仅直接参与骨形成，同时也参与调控破骨细胞功能而影响骨吸收。成骨细胞增殖、分化过程受遗传因素、激素水平及细胞调控因子的影响与调控，其增殖、分化缺陷是骨质疏松症发病的重要原因，有大量研究表明改善成骨细胞功能可治疗骨质疏松症。

 

摘要
背景：骨在一生中不断地进行重塑，骨稳态是成骨细胞的骨形成和破骨细胞的骨吸收之间的动态平衡。在细胞水平严格控制骨重建，对于维持骨稳态以响应结构和代谢需求是至关重要的。
目的：探讨罗汉果皂苷V对成骨细胞增殖、分化的影响。
方法：采用两步酶消化法提取大鼠成骨细胞，通过MTT实验检测罗汉果皂苷V的细胞毒性；不同质量浓度(0，6.25×10-3，1.25×10-2，2.5×10-2 g/L)罗汉果皂苷V对成骨细胞干预2，4，6 d后通过FDA/PI荧光染色观察成骨细胞活性，干预6 d后通过碱性磷酸酶染色、茜素红染色及qRT-PCR检测观察罗汉果皂苷V对成骨细胞分化的影响。
结果与结论：①MTT实验表明罗汉果皂苷V在6.25×10-3-2.5×10-2 g/L时显著促进成骨细胞增殖；②FDA/PI荧光染色显示6.25×10-3，1.25×10-2，2.5×10-2 g/L罗汉果皂苷V可提高成骨细胞活力、促进细胞增殖，尤其以1.25×10-2 g/L效果显著；③罗汉果皂苷V干预6 d后，碱性磷酸酶染色(定量)及茜素红染色实验均显示罗汉果皂苷V促进成骨细胞成骨矿化，特别是在1.25×10-2 g/L时最为明显；④qRT-PCR结果显示罗汉果皂苷V组的成骨分化相关基因RUNX2、BSP、OCN和COL1A1表达高于对照组，特别是罗汉果皂苷V在1.25× 10-2 g/L时表达最高；⑤结果表明，罗汉果皂苷V可刺激成骨细胞增殖、分化，在1.25×10-2 g/L时作用最为明显。


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: 0000-0001-8821-633X(姚顺晗)

关键词: 罗汉果皂苷V, 成骨细胞, 细胞增殖, 细胞分化, 骨形成, 骨质疏松, 广西自然科学基金

Abstract:

BACKGROUND: Bone remodeling is going on throughout the lifetime. Bone homeostasis is a dynamic balance between bone formation of osteoblasts and bone resorption of osteoclasts. Controlling of bone remodeling at the cellular level is critical to maintaining bone homeostasis in response to structural and metabolic demands.
OBJECTIVE: To explore the effect of mogroside V on the proliferation and differentiation of osteoblasts. 
METHODS: Rat osteoblasts were extracted by two-step enzymatic digestion, followed by MTT assay to detect the toxicity of mogroside V. The proliferation of osteoblasts was observed by FDA/PI fluorescence staining at 2, 4 and 6 days after the treatment with mogroside V (0, 6.25×10-3, 1.25×10-2, 2.5×10-2 g/L). After 6 days of treatment on osteoblasts, the effect of mogroside V on osteoblast differentiation was studied by alkaline phosphatase staining, alizarin red staining and qRT-PCR.
RESULTS AND CONCLUSION: MTT assay showed that the mogroside V at 6.25×10-3 to 2.5×10-2 g/L promoted osteoblast proliferation. FDA/PI fluorescence staining showed that mogroside V (6.25×10-3, 1.25×10-2, 2.5×10-2 g/L) could improve osteoblast activity and promote cell proliferation, especially when the concentration of mogroside V was 1.25×10-2 g/L. After 6 days of treatment, alkaline phosphatase staining and alizarin red staining showed that the mogroside V, especially at 1.25×10-2 g/L, promoted osteogenesis and mineralization. qRT-PCR results showed that the expressions of runt related transcription factor 2, bone sialoprotein, osteocalcin and alpha-1 type I collagen genes were higher in the mogroside V group than the blank group, especially when the concentration of mogroside V was 1.25×10-2 g/L. Therefore, mogroside V, especially at 1.25×10-2 g/L, promotes bone formation by stimulating osteoblast proliferation and differentiation.

Key words: mogroside V, osteoblasts, cell proliferation, cell differentiation, bone formation, osteoporosis, Guangxi Natural Science Foundation

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