中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (21): 3299-3305.doi: 10.3969/j.issn.2095-4344.2017.21.004

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

小分子水凝胶对骨髓间充质干细胞增殖、凋亡及向心肌细胞分化的影响

陈国钦1，黎锦亮1，宋明才1，区彩文2   

  1. 1广州市番禺区中心医院心内科，番禺区心血管病研究所，广东省广州市 511400；2南方医科大学，广东省广州市 510515
  • 修回日期:2017-02-16 出版日期:2017-07-28 发布日期:2017-08-02
  • 通讯作者: 区彩文，副研究员，博士生导师，南方医科大学，广东省广州市 510515
  • 作者简介:陈国钦，男，1978年生，广东省梅州市人，汉族，硕士，副主任医师，主要从事心肌干细胞研究。
  • 基金资助:

    国家自然科学基金青年项目(31400858)：姜黄素多肽小分子水凝胶的制备及性能研究；广东省自然科学基金(2014A030313707)：姜黄素小分子水凝胶的制备及对心肌缺血再灌注的保护作用

Effect of small molecule hydrogels on proliferation, apoptosis and myocardial differentiation of bone marrow mesenchymal stem cells

Chen Guo-qin1, Li Jin-liang1, Song Ming-cai1, Ou Cai-wen2   

  1. 1Department of Cardiology, Central Hospital of Panyu District, Cardiovascular Institution of Panyu District, Guangzhou 5111400, Guangdong Province, China; 2Southern Medical University, Guangzhou 510515, Guangdong Province, China
  • Revised:2017-02-16 Online:2017-07-28 Published:2017-08-02
  • Contact: Ou Cai-wen, Associate researcher, Doctoral supervisor, Southern Medical University, Guangzhou 510515, Guangdong Province, China
  • About author:Chen Guo-qin, Master, Associate chief physician, Department of Cardiology, Central Hospital of Panyu District, Cardiovascular Institution of Panyu District, Guangzhou 5111400, Guangdong Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 31400858; the Natural Science Foundation of Guangdong Province, No. 2014A030313707

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摘要：

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文题释义：
小分子水凝胶与骨髓间充质干细胞增殖：小分子水凝胶有助于细胞抵抗缺氧微环境，提高其存活能力，原因可能是小分子水凝胶提供的三维微环境机械刺激活化骨髓间充质干细胞表面受体，激活细胞内信号途径，直接促进骨髓间充质干细胞的存活。有报道显示，小分子水凝胶也可间接通过某些特异性配体与骨髓间充质干细胞表面受体结合，激活下游信号传导通路，发挥保护细胞抵抗缺血缺氧的作用。另外，小分子水凝胶还可能通过影响骨髓间充质干细胞的旁分泌，间接促进骨髓间充质干细胞的存活。

 

摘要
背景：前期研究研制的一种短肽小分子水凝胶，在改善局部微环境、携带生物活性物质及干预干细胞信号转导途径等方面较其他水凝胶有着更明显的优势。
目的：探讨小分子水凝胶对骨髓间充质干细胞增殖、凋亡、向心肌细胞分化等的影响。
方法：①取第9代SD大鼠骨髓间充质干细胞，分2组培养，实验组加入小分子水凝胶溶液，对照组常规培养，检测培养7 d内的细胞增殖与凋亡；将两组细胞分别置于缺氧环境中孵育12 h，检测细胞凋亡及凋亡蛋白Bcl-2、Bax、Caspase-3的表达；②取第9代骨髓间充质干细胞，分4组干预：5-氮杂胞苷组加入5-氮杂胞苷，凝胶组加入小分子水凝胶，实验组加入小分子水凝胶与5-氮杂胞苷，正常对照组加入L-DMEM基本培养基，诱导4周后，检测cTnT、desmin及Cx-43蛋白的表达，以及心肌早期转录因子NKX2.5、GATA-4的表达。
结果与结论：①与对照组相比，实验组骨髓间充质干细胞有更好的增殖能力及更低的凋亡数量；在缺氧环境中，实验组和对照组存活的细胞均有减少，实验组的凋亡或坏死细胞数目明显少于对照组(P < 0.05)；实验组细胞Bcl-2蛋白表达高于对照组(P < 0.01)，Bax、Caspase-3蛋白表达低于对照组(P < 0.01)；②5-氮杂胞苷组和实验组各样本中均出现明确的心肌早期分化基因，实验组NKx2.5和GATA-4 mRNA表达高于5-氮杂胞苷组(P < 0.05)。正常对照组cTnT表达阴性，desmin及Cx-43蛋白表达很弱；实验组cTnT、desmin及Cx-43蛋白表达强于5-氮杂胞苷组、凝胶组，凝胶组cTnT、desmin及Cx-43蛋白表达与5-氮杂胞苷组相似；③结果表明，小分子水凝胶作为培养基质，更有利于骨髓间充质干细胞的增殖，减少其凋亡并促进其向心肌细胞分化。

 

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: 0000-0001-8035-544X(区彩文)

关键词: 干细胞, 骨髓干细胞, 小分子水凝胶, 骨髓间充质干细胞, 增殖, 凋亡, 诱导分化, 国家自然科学基金

Abstract:

BACKGROUND: A short-peptide small molecule hydrogel (SMH) developed in the previous study has more obvious advantages than other hydrogels to improve local microenvironment, carry bioactive substances and interfere with stem cell signal transduction pathways.
OBJECTIVE: To explore the effect of SMHs on bone marrow mesenchymal stem cells (BMSCs) proliferation, apoptosis and differentiation into myocardial cells.
METHODS: (1) Passage 9 rat BMSCs in vitro were divided into control group and experimental group, followed by routine culture and culture in SMHs, respectively. At 7 days of culture, cell proliferation and apoptosis were detected. Cells in the two groups were exposed to anaerobic environment for 12 hours, and expression levels of Bcl-2, Bax and Caspase-3 in BMSCs were detected. (2) Passage 9 BMSCs were divided into four groups and then cultured in 5-azacytidine, SMHs, SMHs+5-azacytidine, and L-DMEM (normal control), respectively. After 4 weeks of induction, expression of CTnT, desmin and Cx-43 proteins was detected and expression levels of early cardiac transcription factors, NKX2.5 and GATA-4, were also measured.
RESULTS AND CONCLUSION: (1) Compared with the control group, better proliferation and lower apoptosis of BMSCs were found in the experimental group. Under anaerobic conditions, the number of survival cells was reduced in both groups, but less apoptosis or necrosis was found in the experimental group than the control group (P < 0.05). Moreover, the level of Bcl-2 was higher in the experimental group than the control group (P < 0.01), while the levels of Bax and Caspases-3 protiens were lower in the experimental group than the control group (P < 0.01). (2) NKx2.5 and GATA-4 mRNA expression was found in both 5-azacytidine and SMHs+5-azacytidine groups, and moreover, the mRNA levels of early cardiac transcription factors were significantly higher in the SMHs+5-azacytidine group than in the 5-azacytidine group (P < 0.05). In the normal control group, cTnT expressed negatively, and desmin and Cx-43 expressed weakly. The expression of cTnT, desmin and Cx-43 proteins was higher in the SMHs+5-azacytidine group than in the 5-azacytidine and SMHs groups, while there was no significant difference between the latter two groups. To conclude, SMHs as a culture medium is conducive to the proliferation of BMSCs, reduces cell apoptosis, and promotes myocardial differentiation of BMSCs.

 

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: Hydrogels, Mesenchymal Stem Cells, Cell Proliferation, Apoptosis, Tissue Engineering

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