大鼠骨髓间充质干细胞的培养和转染及其对肠瘘瘘口的影响

发布时间：2018-02-03 07:03

本文关键词： HO-1基因肠瘘细胞移植骨髓间充质干细胞　出处：《山东大学》2010年硕士论文　论文类型：学位论文

【摘要】： 目的：肠瘘是一种严重的术后并发症且死亡率高。主要表现为：瘘口形成与肠内容物漏出、感染、营养不良、水电解质和酸碱平衡紊乱以及多器官功能障碍等。营养支持、抗感染、减少肠液分泌、封堵瘘管、维持内环境稳定、促进瘘管愈合以及选择性手术治疗等综合措施,可以提高治愈率。血红素加氧酶-Ⅰ(HO-1)也称为热休克蛋白32,是血红素降解过程中的限速酶,它可以降解衰老或破损红细胞释放出的血红素,生成胆绿素、胆红素、一氧化碳(CO)和铁蛋白。大量事实证明,HO-1在保持细胞稳态方面能起到极其重要的作用,可以减轻由免疫介导、氧化应激、炎性细胞浸润导致的组织损伤。腺相关病毒(adeno-associated virus,AAV)是安全有效的基因载体,且有外源基因可以长期稳定表达,分裂期细胞和静止期细胞均能被感染等优点,还能将基因转人非周期(noncyling)肿瘤细胞。骨髓间充质干细胞属于多潜能干细胞,分化潜能更广,具有自我更新能力及多向分化能力。能在合适的分化条件下不仅可以分化为成骨细胞、脂肪细胞、软骨细胞、肌肉细胞等中胚层细胞,而且可以分化为外胚层和内胚层的细胞。它成为多种组织修复的种子细胞与基因治疗的良好载体。在严重的组织缺血和损伤的病例中可观察到骨髓间充质干细胞迁徙至损伤部位,并分泌生物活性因子,促进修复与再生。骨髓间充质干细胞可以通过骨髓迁徙或体外扩增后直接或全身注射到达损伤部位,抑制疤痕与凋亡,促进血管生成及祖细胞分裂分化,有利于损伤的愈合。本文探讨转染HO-1基因的骨髓间充质干细胞移植对大鼠肠瘘模型的影响。方法：骨髓间充质干细胞的分离与培养。骨髓间充质干细胞通过腺相关病毒介导转染H0-1基因。以Wistar大鼠建立肠瘘模型,实验动物随机分为实验组(术后24小时造瘘口注射1×106个转染目的基因的骨髓干细胞),,对照组(术后24小时造瘘口注射生理盐水100ul)。移植70天后观察瘘口愈合情况。结果：实验组瘘口愈合明显快于对照组。结论：转染H0-1基因的骨髓间充质干细胞移植有利于大鼠肠瘘模型瘘口的愈合。
[Abstract]:Objective: intestinal fistula is a serious postoperative complication with high mortality. The main manifestations are fistula formation, leakage of intestinal contents, infection and malnutrition. Water, electrolyte, acid-base balance disorder and multiple organ dysfunction. Nutritional support, anti-infection, reduce intestinal secretion, close fistula, maintain the stability of the internal environment. Heme oxygenase-I (HO-1), also known as heat shock protein 32 (HSP32), is a rate-limiting enzyme in heme degradation. It degrades heme released by aging or damaged red blood cells and produces bilirubin, carbon monoxide (CO) and ferritin. HO-1 can play an extremely important role in maintaining the homeostasis of cells and can alleviate the immune-mediated oxidative stress. Adeno-associated virus (AAVV) is a safe and effective gene vector. Moreover, it has the advantages of long-term stable expression of exogenous genes and infection of both mitotic and stationary cells. Bone marrow mesenchymal stem cells (BMSCs) are multipotent stem cells and have a wider differentiation potential. It can not only differentiate into osteoblasts, adipocytes, chondrocytes, muscle cells and other mesoderm cells, but also can differentiate into osteoblasts, adipocytes, chondrocytes, muscle cells and so on. And it can differentiate into ectodermal and endoderm cells. It is a good carrier of seed cells and gene therapy for multiple tissue repair. Bone marrow mesenchymal stem cells can be observed in severe tissue ischemia and injury cases. Migrate to the site of the injury. Bone marrow mesenchymal stem cells (BMSCs) can be directly or completely injected into the injured site after bone marrow migration or expansion in vitro to inhibit scar and apoptosis. The effects of bone marrow mesenchymal stem cell transplantation (BMSCs) transfected with HO-1 gene on intestinal fistula in rats were studied. Methods: bone marrow mesenchymal stem cells were isolated and cultured. Bone marrow mesenchymal stem cells were transfected with H0-1 gene mediated by adeno-associated virus. Intestinal fistula model was established in Wistar rats. The experimental animals were randomly divided into experimental group (1 脳 106 bone marrow stem cells transfected with target gene) were injected 24 hours after operation. The control group (24 hours after operation) was injected with normal saline 100 ulus. The healing of fistula was observed 70 days after transplantation. Results: the healing of fistula in the experimental group was faster than that in the control group. Conclusion: transplantation of bone marrow mesenchymal stem cells transfected with H0-1 gene is beneficial to the healing of fistula in rat intestinal fistula model.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

【参考文献】