汗腺样细胞复合羊皮脱细胞真皮基质对创面愈合影响的研究

发布时间：2018-03-14 13:09

  本文选题：汗腺样细胞　切入点：骨髓间充质干细胞　出处：《内蒙古大学》2017年硕士论文　论文类型：学位论文

【摘要】：皮肤是机体最大的器官,作为机体最表层,皮肤像一道屏障覆于机体的表面,起到保持皮肤水分,维持正常生理功能的作用。当皮肤受到外界损伤或者疾病等因素的影响而造成缺损时,会对生命体造成严重的危害,甚至危及生命。研究发现,细胞治疗有助于皮肤损伤的修复。本研究主要探讨由大鼠骨髓间充质干细胞(Rat Bone marrow mesenchymal stem cells,rBM-MSCs)诱导的汗腺样细胞(sweat gland-like cells)与羊脱细胞真皮基质(acellular dermal matrix,ADM)复合后对创面愈合的影响。首先,本研究以大鼠骨髓为材料来源,采用全骨髓贴壁法培养获得目的细胞。通过形态、增殖能力和表面标志的鉴定显示出所得细胞为rBM-MSCs。其次,通过将rBM-MSCs与大鼠热休克汗腺细胞(sweat glands cells,SGCs)共培养的方法进行细胞诱导。RT-PCR和细胞免疫荧光检测显示:诱导所得细胞为汗腺样细胞。随后,采用酶消化法制备出羊ADM,通过MTS法检测细胞毒性和HE染色检测其脱细胞程度显示:ADM脱细胞程度较为彻底,且没有细胞毒性。最后,分别将复合有汗腺样细胞的ADM和未复合汗腺样细胞的ADM覆盖于裸鼠损伤创面上。通过对创面愈合率、参与创面愈合的蛋白VEGF和Collagen-Ⅰ的表达情况、愈合创面细胞凋亡相关基因Bax/Bcl-2的相对表达量的变化的测定显示:汗腺样细胞复合羊ADM抑制了愈合创面组织的细胞凋亡并且加快了创面的愈合速率。综上所述,由BM-MSCs诱导分化的汗腺样细胞与羊ADM复合后抑制了愈合创面组织的细胞凋亡并且加快了创面的愈合速率。本研究初步探讨了大鼠汗腺样细胞复合羊ADM对于创面愈合的影响,为后续深入研究其作用机制提供了一定的依据。
[Abstract]:The skin is the largest organ of the body. As the most superficial part of the body, the skin is like a barrier covering the surface of the body, which helps to maintain the moisture of the skin. The function of maintaining normal physiological function. When the skin is damaged by external injury or disease and other factors, it will cause serious harm to the life body, and even endanger the life. The purpose of this study was to investigate the effects of Bone marrow mesenchymal stem cells (rBM-MSCs) induced by rat bone marrow mesenchymal stem cells (Bone marrow mesenchymal stem cells rBM-MSCs) on wound healing after the combination of sweat gland-like cells and sheep acellular dermal matrix adm. In this study, the target cells were obtained from rat bone marrow by whole bone marrow adherent method. The morphology, proliferative ability and surface markers showed that the cells were rBM-MSCs. By co-culture of rBM-MSCs and sweat glands cells SGCsof rat sweat gland cells under heat shock, the results of cell induction. RT-PCR and immunofluorescence analysis showed that the cells induced were sweat gland like cells. Sheep ADMs were prepared by enzyme digestion. The cytotoxicity was detected by MTS method and the degree of acellular by HE staining showed that the acellular degree of ADM was relatively thorough and had no cytotoxicity. The ADM and ADM of the combined sweat gland like cells and the uncomplex sweat gland like cells were respectively covered on the injured wound of nude mice. The expression of protein VEGF and Collagen- 鈪，

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