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

发布时间：2018-03-29 17:07

本文选题：水凝胶　切入点：间质干细胞　出处：《中国组织工程研究》2017年21期

【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(P0.05);实验组细胞Bcl-2蛋白表达高于对照组(P0.01),Bax、Caspase-3蛋白表达低于对照组(P0.01);(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(P0.05)。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。
[Abstract]:Background: a small molecular hydrogel with short peptides was developed in previous studies to improve the local microenvironment. Carrying bioactive substances and interfering with signal transduction pathway of stem cells have more obvious advantages than other hydrogels. Objective: to investigate the proliferation and apoptosis of bone marrow mesenchymal stem cells by small molecule hydrogel. Methods Sprague-Dawley rat bone marrow mesenchymal stem cells (BMSCs) of the 9th generation were cultured in two groups. The experimental group was cultured with small molecular hydrogel solution and the control group was cultured routinely. The proliferation and apoptosis of the cells were detected within 7 days of culture. The two groups were incubated in anoxic environment for 12 hours respectively. Apoptosis and the expression of apoptotic protein Bcl-2nBaxCaspase-3 were detected. Bone marrow mesenchymal stem cells (BMSCs) of the 9th generation were collected. The cells were divided into 4 groups and treated with the addition of 5-azacytidine to the control group. Small molecule hydrogels were added to the gel group, small molecule hydrogels and 5-azacytidine were added to the experimental group, and the normal control group was added to the basic L-DMEM medium. After 4 weeks of induction, the expression of ct nTtdesmin and Cx-43 protein were detected. Results and conclusion compared with control group, bone marrow mesenchymal stem cells in experimental group had better proliferative ability and lower apoptotic quantity. The number of surviving cells decreased in both experimental and control groups. The number of apoptotic or necrotic cells in the experimental group was significantly lower than that in the control group (P 0.05), the expression of Bcl-2 protein in the experimental group was higher than that in the control group (P 0.01) and the expression of Caspase-3 was lower than that in the control group (P 0.01). The expression of NKx2.5 and GATA-4 mRNA in the experimental group was higher than that in the 5-azacytidine group (P 0.05). The expression of cTnT and Cx-43 protein in the normal control group was very weak, and the expression of ct nTnTnttdesmin and Cx-43 protein in the experimental group was stronger than that in the 5-azacytidine group, and the expression of Cx-43 protein in the gel group was higher than that in the control group. The results showed that the small molecular hydrogel was more favorable to the proliferation of bone marrow mesenchymal stem cells, reduced its apoptosis and promoted its differentiation into cardiomyocytes.
【作者单位】：广州市番禺区中心医院心内科番禺区心血管病研究所;南方医科大学;
【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~~
【分类号】：R318

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