脂肪源间充质干细胞对小鼠急性肾损伤后干细胞因子及其受体表达的影响

发布时间：2018-03-22 17:14

本文选题：脂肪源间充质干细胞　切入点：c-KIT　出处：《新乡医学院》2014年硕士论文　论文类型：学位论文

【摘要】：背景急性肾损伤(Acute kidney injury, AKI)是一种常见危急重症。多种原因可造成AK_I,如重症感染及药物等。随着诊疗技术水平的提高,AKI的病死率有所下降,但是在合并AKI重症病人的病死率仍达65%。目前AKI的治疗以肾脏替代治疗为主。近年脂肪源间充质干细胞(adipose-derived mesenchymal stem cells, ADMSCs)治疗AKI成为肾脏病工作者研究热点之一。研究表明ADMSCs具有治疗AKI的作用,且其来源丰富、易于分离培养、免疫原性低、体外扩增快等优点。但其具体的治疗机制仍不确切。目的 1.通过观察肾损伤标志物,来判断ADMSCs对急性肾损伤的治疗作用。 2.通过测定相关因子来分析ADMSCs治疗小鼠AKI可能的机制,以期为临床应用ADMSCs治疗AKI提供有力的实验依据。方法 1.30只雄性KM小鼠随机分成3组,正常对照(normal control)组、模型组(model group)和治疗组(treatment group),每组10只小鼠。 2.正常组不做任何干预措施,模型组、治疗组分别腹腔注射顺铂,治疗组于注射顺铂12小时后经尾静脉注射ADMSCs悬液5×105/0.5mL,模型组于注射顺铂12小时后经尾静脉注射等剂量的生理盐水。 3.注射顺铂7天后处死小鼠,心脏取血,测定血清尿素氮(BUN)和血肌酐(Scr)值来评价肾功能情况；通过光学显微镜观察小鼠肾组织病理形态改变；免疫组织化学(immunohistochemistry, IHC)法检测中性粒细胞明胶酶相关脂质运载蛋白(NGAL)、干细胞因子受体(c-KIT)、及干细胞因子(SCF)。结果 1.与模型组相比,治疗组BUN、Scr和NGAL等肾损伤标志物明显下降,及肾小管坏死(acute tubular necrosis, ATN)评分显著降低(P0.05)亦明显下降。 2.与模型组相比,治疗组肾组织中c-KIT、SCF表达明显增加；与正常组相比,模型组肾组织中c-KIT、SCF表达亦有所增加(P0.05)。结论 1.外源性ADMSCs可以有效促进受损肾小管上皮细胞修复的作用； 2.在AKI模型中,外源性ADMSCs可能通过旁分泌或其自身分泌细胞因子SCF及c-KIT的方式,并且激活SCF/c-KIT信号传导通路从而抑制肾小管上皮细胞凋亡并促进残余的肾小管上皮细胞分化、增殖,进而起到保护肾脏作用。
[Abstract]:Background Acute kidney injury (AKI) is a common type of acute acute renal injury. It can be caused by a variety of causes, such as severe infection and drugs, etc. With the improvement of diagnostic and treatment techniques, the mortality of AKI has decreased. However, the mortality rate of severe patients with AKI is still 65%. At present, renal replacement therapy is the main treatment of AKI. Recently, adipose-derived mesenchymal stem cells (ADMSCs) treatment of adipose-derived mesenchymal stem cells (ADMSCs) has become one of the hotspots in the study of renal disease workers. Ming ADMSCs has the effect of treating AKI. Its source is rich, easy to isolate and culture, low immunogenicity, rapid expansion in vitro, but its specific treatment mechanism is still not accurate. Purpose. 1. To evaluate the therapeutic effect of ADMSCs on acute renal injury by observing the markers of renal injury. 2. The possible mechanism of ADMSCs in the treatment of AKI in mice was analyzed by measuring the relevant factors, in order to provide an effective experimental basis for the clinical application of ADMSCs in the treatment of AKI. Method. 1.Thirty male km mice were randomly divided into 3 groups, normal control group, model group (n = 10) and treatment group (n = 10). 2.The normal group did not do any intervention, the model group and the treatment group were intraperitoneally injected with cisplatin. In the treatment group, 5 脳 105% 0.5 mL ADMSCs suspension was injected through the tail vein 12 hours after the injection of cisplatin, while in the model group, the same dose of normal saline was injected through the tail vein 12 hours after the injection of cisplatin. 3. The mice were sacrificed 7 days after injection of cisplatin, the blood was taken from the heart, the serum urea nitrogen (bun) and serum creatinine (SCR) were measured to evaluate the renal function, and the pathological changes of renal tissue were observed by optical microscope. Immunohistochemical method was used to detect neutrophil gelatinase-associated lipid carrier protein (NGALN), stem cell factor receptor c-KITT, and stem cell factor (SCF). Results. 1. Compared with the model group, the renal injury markers such as bun SCR and NGAL in the treatment group were significantly decreased, and the acute tubular necrosis (ATN) score in the treatment group was significantly lower than that in the control group (P 0.05). 2.Compared with the model group, the expression of c-KIT-SCF in the renal tissue of the treatment group was significantly increased, and the expression of c-KIT-SCF in the renal tissue of the model group was also increased compared with the normal group. Conclusion. 1. Exogenous ADMSCs can effectively promote the repair of damaged renal tubular epithelial cells; 2. In AKI model, exogenous ADMSCs may inhibit the apoptosis of renal tubular epithelial cells and promote the differentiation of residual tubular epithelial cells by paracrine or autocrine cytokines SCF and c-KIT, and activate the SCF/c-KIT signal transduction pathway. Proliferation, which in turn plays a role in protecting the kidney.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692.5

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