炎性因子诱导的自噬调控人胎盘MSCs对急性肺损伤治疗的影响

发布时间：2018-06-17 09:33

本文选题：炎性因子 + 间充质干细胞　；参考：《宁夏医科大学》2017年硕士论文

【摘要】：急性肺损伤(ALI)的发病机制十分复杂,现在没有特效的治疗方案,间充质干细胞(MSCs)作为具有自我更新潜能和多向分化潜能等多功能的前体细胞,给ALI的治疗带来一条新思路。间充质干细胞的细胞特性和功能具有一定的可塑性,即随所处微环境的不同而改变,致使其生物学功能在基础研究和临床应用中表现出复杂性和多样性的特点。间充质干细胞在急性肺损伤炎性环境下发挥什么样的细胞特性和功能仍有待研究。目的:研究在炎性环境下的MSCs发生自噬对急性肺损伤治疗的影响。方法:(1)从宁夏医科大学总医院获取健康胎盘,根据已建立的人胎盘来源间充质干细胞分离培养的方法,培养鉴定间充质干细胞。(2)应用IL-1β、IL-6、IFN-γ和TNF-α炎症因子处理人胎盘来源间充质干细胞,western blot检测自噬标记蛋白LC3I/II、Atg5和Atg7表达情况;同时观察同等处理条件下间充质干细胞的增殖情况以及自噬抑制剂3-MA处理后的增殖情况。(3)应用western blot检测凋亡标志因子Caspase3、PARP1。(4)利用博来霉素(BLM)制备小鼠急性肺损伤模型,尾静脉注射感染si-NC和si-Atg7慢病毒的人胎盘来源间充质干细胞,观察发生自噬和不发生自噬的间充质干细胞在小鼠急性肺损伤模型中的存活率;观察间充质干细胞干预后急性肺损伤小鼠的肺组织病理改变。结果:(1)我们成功分离、培养了人胎盘来源间充质干细胞。(2)炎症因子IL-1β、IL-6、IFN-γ和TNF-α可诱导间充质干细胞发生自噬;其中,IFN-γ和TNF-α诱导的自噬抑制间充质干细胞增殖;抑制IFN-γ和TNF-α诱导的自噬能够恢复间充质干细胞的增殖。(3)IFN-γ、TNF-α诱导间充质干细胞发生自噬可导致其凋亡。(4)在博来霉素(BLM)诱导的小鼠急性肺损伤模型中,si-Atg7组较si-NC组,间充质干细胞存活率更高,肺组织病理改善明显。结论:(1)炎性因子IL-1β、IL-6、IFN-γ和TNF-α可诱导间充质干细胞发生自噬。(2)IFN-γ和TNF-α可诱导间充质干细胞发生自噬并导致其凋亡,降低了间充质干细胞的存活率,影响其对急性肺损伤的治疗作用。
[Abstract]:The pathogenesis of acute lung injury (Ali) is very complicated. There is no special therapeutic scheme now. Mesenchymal stem cells (MSCs), as multifunctional progenitor cells with self-renewal potential and multi-differentiation potential, bring a new thought to the treatment of Ali. The characteristics and functions of mesenchymal stem cells have a certain degree of plasticity, that is, the biological functions of mesenchymal stem cells vary with the microenvironment, which leads to the complexity and diversity of their biological functions in basic research and clinical application. The characteristics and functions of mesenchymal stem cells in inflammatory environment of acute lung injury remain to be studied. Objective: to study the effect of autophagy of MSCs in inflammatory environment on the treatment of acute lung injury. Methods healthy placenta was obtained from the General Hospital of Ningxia Medical University, and the human placenta derived mesenchymal stem cells were isolated and cultured according to the established method of isolation and culture of human placenta derived mesenchymal stem cells. Cultured and identified mesenchymal stem cells (MSCs) were treated with IL-1 尾 IL-6 IFN- 纬 and TNF- 伪 inflammatory cytokines. Western blot was used to detect the expression of autophagy marker protein LC3I / III-Atg5 and Atg7 in human placenta derived mesenchymal stem cells. At the same time, the proliferation of mesenchymal stem cells under the same conditions and the proliferation after treatment with autophagy inhibitor 3-MA were observed. The mouse model of acute lung injury was established with bleomycin (BLM). Human placental mesenchymal stem cells (MSCs) infected with si-NC and si-Atg7 lentivirus were injected via tail vein to observe the survival rate of mesenchymal stem cells (MSCs) with or without autophagy in mice with acute lung injury. To observe the pathological changes of lung tissue in mice with acute lung injury after the intervention of mesenchymal stem cells. Results: we successfully isolated and cultured human placental derived mesenchymal stem cells. 2) the inflammatory cytokines IL-1 尾, IL-6, IFN- 纬 and TNF- 伪 could induce autophagy of mesenchymal stem cells, in which IFN- 纬 and TNF- 伪 induced autophagy inhibited the proliferation of mesenchymal stem cells. Inhibition of IFN- 纬 and TNF- 伪 induced autophagy could restore the proliferation of mesenchymal stem cells. The survival rate of mesenchymal stem cells was higher and the pathological changes of lung tissue were obvious. Conclusion Interleukin-1 尾 IL-6 IFN- 纬 and TNF- 伪 can induce autophagy and apoptosis of mesenchymal stem cells, decrease the survival rate of mesenchymal stem cells and affect their therapeutic effect on acute lung injury.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R563.8

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