不同浓度牡蛎蛋白肽对大鼠骨髓间充质干细胞增殖和分化能力的影响

doi:10.19438/j.cjoms.2021.01.002

中国口腔颌面外科杂志 ›› 2021, Vol. 19 ›› Issue (1): 6-11.doi: 10.19438/j.cjoms.2021.01.002

不同浓度牡蛎蛋白肽对大鼠骨髓间充质干细胞增殖和分化能力的影响

王毅炜¹, 李晨琳², 蒋倩^2,*, 徐光宙^2,*

1.淮安市口腔医院口腔内科,江苏淮安 223300;
2.上海交通大学医学院附属第九人民医院·口腔医学院口腔外科,国家口腔疾病临床医学研究中心, 上海市口腔医学重点实验室,上海市口腔医学研究所,上海 200011

收稿日期:2020-03-06 修回日期:2020-06-11 出版日期:2021-01-20 发布日期:2021-02-19
通讯作者: 蒋倩,E-mail: jiangqian0720@hotmail.com;徐光宙,E-mail: xgzmy@163.com。^*共同通信作者
作者简介:王毅炜（1988-）,男,主治医师,E-mail: wangyiwei218@163.com
基金资助:
国家自然科学基金（31800816）; 浦东新区卫生和计划生育委员会卫生计生科研项目（PW2018D-08）; 上海申康医院发展中心临床科技创新项目-市级医院新兴前沿技术联合攻关项目（SHDC12019X01）; 上海交通大学医学院附属第九人民医院创客基金（CK2018002）

Effects of oyster protein peptide with different concentrations on the proliferation and differentiation of bone marrow mesenchymal stem cells

WANG Yi-wei¹, LI Chen-lin², JIANG Qian², XU Guang-zhou²

1. Department of Oral Medicine, Huaian Stomatological Hospital. Huaian 223300, Jiangsu Province;
2. Department of Oral Surgery, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology. Shanghai 200011, China

Received:2020-03-06 Revised:2020-06-11 Online:2021-01-20 Published:2021-02-19

摘要/Abstract

摘要： 目的: 探讨不同浓度牡蛎蛋白肽对骨髓间充质干细胞（bone marrow mesenchymal stem cells, BMSCs）成骨分化的影响。方法: 选择具备稳定传代能力的第3 代大鼠BMSCs,采用MTT试剂盒,检测细胞在0、10、20、50 mg/L 不同牡蛎蛋白肽浓度干预培养24 h、3 d、7 d后的细胞活力。然后将BMSCs 分为普通培养基组（DMEM）和成骨诱导培养基组（ODM）,根据牡蛎蛋白肽的不同浓度,将实验组分为4组,依次为0、10、20、50 mg/L,利用碱性磷酸酶（ALP）染色法和ALP半定量检测法,研究不同浓度牡蛎蛋白肽对BMSCs 培养10 d 早期成骨分化的影响。应用茜素红染色法,半定量检测不同浓度牡蛎蛋白肽对BMSCs 培养21 d 后成骨矿化的影响,应用RT-PCR检测成骨相关基因的表达情况。采用SPSS 20.0 软件包对数据进行统计学分析。结果: MTT结果表明,经不同浓度药物干预处理后,细胞活力旺盛,无明显异常;但牡蛎蛋白肽药物浓度为50 mg/L 时,细胞死亡数有上升趋势。ALP半定量检测结果显示,DMEM和ODM 组诱导分化第10 天时,20 mg/L 浓度组的ALP 活性显著高于对照组（P<0.05）,50 mg/L 浓度组的ALP活性与对照组无显著差异。茜素红半定量检测结果显示,ODM 组中20 mg/L 实验组的A值显著高于对照组（P<0.05）。与对照组及50 mg/L组相比,10、20 mg/L组RUNX-2和OCN的表达显著上升（P<0.05）。结论: 高于50 mg/L 浓度的牡蛎蛋白肽对BMSCs 的成骨分化表现出明显抑制作用,而低浓度牡蛎蛋白肽,尤其是20 mg/L 的牡蛎蛋白肽对BMSCs 成骨分化及细胞生长有促进作用。

关键词: 牡蛎蛋白肽, 干细胞, 成骨分化, 中药

Abstract: PURPOSE: To investigate the effect of different concentrations of oyster protein peptide on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). METHODS: BMSCs of the third generation from rats with stable subculture ability were selected, and the cell viability was detected by MTT kit at different concentrations of 0, 10, 20 and 50 mg/L oyster protein peptide for 24 hours, 3 days and 7 days. BMSCs were divided into two groups: cultured with DMEM or ODM. According to the different concentrations of oyster protein peptide, the experimental groups were divided into four subgroups, which were 0, 10, 20 and 50 mg/L, respectively. The effects of different concentrations of oyster protein peptide on the early osteogenic differentiation of BMSCs were evaluated by ALP staining and ALP semi-quantitative detection. Alizarin red staining was used to detect the effect of different concentrations of oyster protein peptide on the osteogenesis and mineralization of BMSCs after 21 days of treatment. RT-PCR was used to defect the expression of bone formation-related genes. SPSS 20.0 software package was used to analyze the data. RESULTS: The results of MTT showed that cell activity was vigorous and there was no obvious abnormality after intervention with different concentrations of drugs, but the number of cell death increased when the concentration of oyster protein peptide was 50 mg/L. The results of ALP semi-quantitative analysis showed that ALP activity of DMEM and ODM group was significantly higher than that of the control group (P<0.05) on the 10th day of differentiation induction, but there was no significant difference between 50 mg/L group and the control group. Alizarin red semi-quantitative analysis showed that the A value of 20 mg/L in ODM group was significantly higher than that of the control group(P<0.05). The expression of RUNX-2 and OCN in 10 mg/L and 20 mg/L groups were significantly higher than that in control goup and 50 mg/L group(P<0.05). CONCLUSIONS: Oyster protein peptide with a concentration higher than 50 mg/L can significantly inhibit osteogenic differentiation of BMSCs, while oyster protein peptide with a low concentration, especially oyster protein peptide with a concentration of 20 mg/L, can promote osteogenic differentiation and cell growth of BMSCs.

Key words: Oyster protein peptide, Stem cell, Osteogenic differentiation, Traditional Chinese medicine

中图分类号:

王毅炜, 李晨琳, 蒋倩, 徐光宙. 不同浓度牡蛎蛋白肽对大鼠骨髓间充质干细胞增殖和分化能力的影响[J]. 中国口腔颌面外科杂志, 2021, 19(1): 6-11.

WANG Yi-wei, LI Chen-lin, JIANG Qian, XU Guang-zhou. Effects of oyster protein peptide with different concentrations on the proliferation and differentiation of bone marrow mesenchymal stem cells[J]. China J Oral Maxillofac Surg, 2021, 19(1): 6-11.

参考文献

[1] Hench LL, Thompson I Twenty-first century challenges for biomaterials[J]. J R Soc Interface, 2010, 7(Suppl 4): S379-S391.
[2] Naderi H, Matin MM, Bahrami AR.Review paper: critical issues in tissue engineering: biomaterials, cell sources, angiogenesis, and drug delivery systems[J]. J Biomater Appl, 2011, 26(4): 383-417.
[3] 薛晶,俞艳,王利娟,等.中药黄芩苷对牙髓干细胞增殖及成牙成骨分化能力的影响[J].口腔生物医学,2015, 6(1): 15-18.
[4] 葛保健,王业华,郭含军,等.依普黄酮对去卵巢大鼠骨质疏松的防治作用[J].中国新药与临床杂志,2010, 29(2): 94--98.
[5] 林海生,曹文红,章超桦,等.牡蛎蛋白酶解物的制备及其抗氧化活性研究[J]. 食品工业科技, 2013, 34(16): 163-168.
[6] 王长伟,李八方,宋文山,等.牡蛎蛋白肽的制备及促进生长发育功能[J]. 安徽农业科学, 2019, 47(17): 161-164.
[7] Wang X, Zeng D, Weng W, et al.Alendronate delivery on amino . modified mesoporous bioactive glass scaffolds to enhance bone regeneration in osteoporosis rats []. Artif Cells Nanomed . Biotechnol, 2018, 46(Sup2): 171-181.
[8] 陈秋月. 骨髓间充质干细胞与牙髓细胞共培养对细胞增殖、矿化及IL-1β分泌影响的研究[D].广州:南方医科大学, 2015.
[9] 郑奥,曹玲燕,刘阳,等.石墨烯/PLGA复合支架对骨髓间充质干细胞增殖和分化能力的影响[J].口腔生物医学,2018,9(2): 58-62.
[10] Lu H, Kraut D, Gerstenfeld LC, et al. Diabetes interferes with the bone formation by affecting the expression of transcription factors that regulate osteoblast differentiation[J]. Endocrinol, 2003, 144(1): 346--352.
[11] Cognee KG, Doeleman HM, Lalanne P, et al.Cooperative interactions between nano-antennas in a high-Q cavity for unidirectional light sources[J]. Light Sci Appl, 2019, 8: 115-115.

[1]	廖寅秀, 张茂林, 邹多宏. PDGFD对人牙髓干细胞迁移及成牙本质向分化的作用[J]. 中国口腔颌面外科杂志, 2024, 22(5): 417-423.
[2]	袁润林, 李亚明. BMP-2联合雷奈酸锶对大鼠骨髓间充质干细胞成骨分化的影响[J]. 中国口腔颌面外科杂志, 2024, 22(5): 430-434.
[3]	王周阳, 凌霄, 施燕妮, 王磊, 秦兴军. ANO5非移码插入突变型GDD家系临床分析及其诱导多能干细胞的构建[J]. 中国口腔颌面外科杂志, 2024, 22(1): 16-22.
[4]	王周阳，凌霄，施燕妮，王磊，秦兴军. ANO5非移码插入突变型GDD家系临床分析及其诱导多能干细胞的构建[J]. 中国口腔颌面外科杂志, 2024, 22(1): 36-41.
[5]	林惠, 高云飞, 刘晶, 马旭辉. 槐角提取物对颌骨骨髓间充质干细胞成骨向分化的影响[J]. 中国口腔颌面外科杂志, 2024, 22(1): 73-77.
[6]	庞超远, 田文艳. 骨髓移植术后冠周炎致口腔软组织感染1例报告[J]. 中国口腔颌面外科杂志, 2023, 21(4): 421-424.
[7]	蒋胜杰, 梅鹏, 周皓楠, 潘兆澄, 高源, 蔡鸣. 三维打印含铜钛合金对骨髓间充质干细胞成骨分化的影响[J]. 中国口腔颌面外科杂志, 2022, 20(4): 334-339.
[8]	柴盈, 刘芷扬, 刘玥旻, 陈敏洁, 杨驰. 人牙髓干细胞来源外泌体促进周围神经损伤修复的实验研究[J]. 中国口腔颌面外科杂志, 2022, 20(2): 105-110.
[9]	谢昕儒, 冉雪会, 徐伟峰, 刘秀明, 张翔凯, 曹玲燕, 张善勇. 小分子鹰嘴豆芽素A对人牙髓干细胞成骨分化的影响[J]. 中国口腔颌面外科杂志, 2021, 19(3): 205-212.
[10]	支音, 郁爽, 沈洪洲, 司家文, 袁媛, 史俊, 刘昌胜. PEGS/β-TCP屏障膜在促进大鼠颅骨缺损骨组织再生中的作用[J]. 中国口腔颌面外科杂志, 2020, 18(3): 219-225.
[11]	刘国新, 王伟伟, 麦潋曦, 焦九阳, 谢舒乐, 林钊宇. 顺铂诱导舌鳞癌细胞发生上皮-间充质转化及获得肿瘤干细胞样特性的实验研究[J]. 中国口腔颌面外科杂志, 2020, 18(1): 10-16.
[12]	张意闻, 罗远, 郭华艳, 张迎娣, 缪国俊, 黄远亮. 淫羊藿次苷对犬颌骨间充质干细胞增殖与成骨分化的影响[J]. 中国口腔颌面外科杂志, 2019, 17(4): 304-310.
[13]	卢陈佩, 李洪亮, 欧阳宁鹃, 林雨恒, 司家文, 沈国芳. MicroRNA-103-3p对小鼠前成骨细胞MC3T3-E1成骨分化早期的影响[J]. 中国口腔颌面外科杂志, 2018, 16(4): 302-309.
[14]	何倩婷, 汤磊乐, 孙菁菁, 卢志远, 王安训. Bmi1对舌鳞癌侧群细胞迁移、侵袭和增殖能力的调控作用及机制探讨[J]. 中国口腔颌面外科杂志, 2018, 16(2): 102-107.
[15]	张广德，靳霞，李荣亮，杨世茂，郭延伟，房殿吉. BMP-9/EPO双基因共转染ADSCs复合nHAC/PLA支架修复兔下颌骨缺损的实验研究[J]. 中国口腔颌面外科杂志, 2017, 15(3): 202-208.

不同浓度牡蛎蛋白肽对大鼠骨髓间充质干细胞增殖和分化能力的影响

Effects of oyster protein peptide with different concentrations on the proliferation and differentiation of bone marrow mesenchymal stem cells

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价