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cT nT、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);(2)5-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 mR NA表达高于5-氮杂胞苷组(P<0.05)。正常对照组cT nT表达阴性,desmin及Cx-43蛋白表达很弱;实验组cT nT、desmin及Cx-43蛋白表达强于5-氮杂胞苷组、凝胶组,凝胶组cT nT、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。
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 CTn T, 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 mR NA expression was found in both 5-azacytidine and SMHs＋5-azacytidine groups, and moreover, the mR NA 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, cT nT expressed negatively, and desmin and Cx-43 expressed weakly. The expression of cT nT, 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.
出处
《中国组织工程研究》
CAS
北大核心
2017年第21期3299-3305,共7页
Chinese Journal of Tissue Engineering Research
基金
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~~
