本文選題：炎性因子 + 間充質(zhì)干細(xì)胞�。� 參考：《寧夏醫(yī)科大學(xué)》2017年碩士論文

【摘要】：急性肺損傷(ALI)的發(fā)病機(jī)制十分復(fù)雜,現(xiàn)在沒(méi)有特效的治療方案,間充質(zhì)干細(xì)胞(MSCs)作為具有自我更新潛能和多向分化潛能等多功能的前體細(xì)胞,給ALI的治療帶來(lái)一條新思路。間充質(zhì)干細(xì)胞的細(xì)胞特性和功能具有一定的可塑性,即隨所處微環(huán)境的不同而改變,致使其生物學(xué)功能在基礎(chǔ)研究和臨床應(yīng)用中表現(xiàn)出復(fù)雜性和多樣性的特點(diǎn)。間充質(zhì)干細(xì)胞在急性肺損傷炎性環(huán)境下發(fā)揮什么樣的細(xì)胞特性和功能仍有待研究。目的:研究在炎性環(huán)境下的MSCs發(fā)生自噬對(duì)急性肺損傷治療的影響。方法:(1)從寧夏醫(yī)科大學(xué)總醫(yī)院獲取健康胎盤,根據(jù)已建立的人胎盤來(lái)源間充質(zhì)干細(xì)胞分離培養(yǎng)的方法,培養(yǎng)鑒定間充質(zhì)干細(xì)胞。(2)應(yīng)用IL-1β、IL-6、IFN-γ和TNF-α炎癥因子處理人胎盤來(lái)源間充質(zhì)干細(xì)胞,western blot檢測(cè)自噬標(biāo)記蛋白LC3I/II、Atg5和Atg7表達(dá)情況;同時(shí)觀察同等處理?xiàng)l件下間充質(zhì)干細(xì)胞的增殖情況以及自噬抑制劑3-MA處理后的增殖情況。(3)應(yīng)用western blot檢測(cè)凋亡標(biāo)志因子Caspase3、PARP1。(4)利用博來(lái)霉素(BLM)制備小鼠急性肺損傷模型,尾靜脈注射感染si-NC和si-Atg7慢病毒的人胎盤來(lái)源間充質(zhì)干細(xì)胞,觀察發(fā)生自噬和不發(fā)生自噬的間充質(zhì)干細(xì)胞在小鼠急性肺損傷模型中的存活率;觀察間充質(zhì)干細(xì)胞干預(yù)后急性肺損傷小鼠的肺組織病理改變。結(jié)果:(1)我們成功分離、培養(yǎng)了人胎盤來(lái)源間充質(zhì)干細(xì)胞。(2)炎癥因子IL-1β、IL-6、IFN-γ和TNF-α可誘導(dǎo)間充質(zhì)干細(xì)胞發(fā)生自噬;其中,IFN-γ和TNF-α誘導(dǎo)的自噬抑制間充質(zhì)干細(xì)胞增殖;抑制IFN-γ和TNF-α誘導(dǎo)的自噬能夠恢復(fù)間充質(zhì)干細(xì)胞的增殖。(3)IFN-γ、TNF-α誘導(dǎo)間充質(zhì)干細(xì)胞發(fā)生自噬可導(dǎo)致其凋亡。(4)在博來(lái)霉素(BLM)誘導(dǎo)的小鼠急性肺損傷模型中,si-Atg7組較si-NC組,間充質(zhì)干細(xì)胞存活率更高,肺組織病理改善明顯。結(jié)論:(1)炎性因子IL-1β、IL-6、IFN-γ和TNF-α可誘導(dǎo)間充質(zhì)干細(xì)胞發(fā)生自噬。(2)IFN-γ和TNF-α可誘導(dǎo)間充質(zhì)干細(xì)胞發(fā)生自噬并導(dǎo)致其凋亡,降低了間充質(zhì)干細(xì)胞的存活率,影響其對(duì)急性肺損傷的治療作用。
[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- 緯 and TNF- 偽 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- 緯 and TNF- 偽 could induce autophagy of mesenchymal stem cells, in which IFN- 緯 and TNF- 偽 induced autophagy inhibited the proliferation of mesenchymal stem cells. Inhibition of IFN- 緯 and TNF- 偽 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- 緯 and TNF- 偽 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.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R563.8

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相關(guān)期刊論文 前2條

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2 Dobroslav Kyurkchiev;Ivan Bochev;Ekaterina Ivanova-Todorova;Milena Mourdjeva;Tsvetelina Oreshkova;Kalina Belemezova;Stanimir Kyurkchiev;;Secretion of immunoregulatory cytokines by mesenchymal stem cells[J];World Journal of Stem Cells;2014年05期

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本文編號(hào)：2030565