单核细胞向淋巴管内皮细胞的诱导分化

发布时间：2018-06-14 06:56

本文选题：单核细胞 + 转分化　；参考：《山东大学》2009年硕士论文

【摘要】： 研究背景:淋巴管新生通常发生在肿瘤、创伤、炎症等许多疾病的病理生理过程中。以往的研究认为,淋巴管新生的途径主要有二:原有的淋巴管在VEGF-C的作用下以出芽方式形成新的淋巴管;外周血内皮祖细胞迁移到局部组织,在VEGF-C的诱导下转分化为淋巴管内皮细胞。在肿瘤、创伤、炎症等病理情况下,单核细胞通过血液循环迁移至局部组织转化为巨噬细胞,通过产生VEGF-C,对淋巴管新生发挥重要作用。目前研究已发现,单核细胞在VEGF等的作用下能够向血管内皮细胞转分化,但能否在某些因子的诱导下转分化为淋巴管内皮细胞,尚未见体外研究证实。本研究选择在炎症等病理状态下组织产生的能活化单核细胞的因子,如:细胞外基质成分纤维连接蛋白(fibronectin,FN)、炎性因子脂多糖(lipopolysaccharide,LPS)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)以及血管内皮生长因子C(vascular endothelial growm factor-C,VEGF-C)、白细胞介素-3(interleukin-3,IL-3)、白细胞介素-7(interleukin-7,IL-7)等,体外诱导单核细胞,检测其淋巴管内皮细胞标志物LYVE-1、Podoplanin、Prox1及内皮细胞标志物vWF和eNOS的表达,观察其向淋巴管内皮细胞转分化的可能性,为进一步研究淋巴管新生机理提供理论依据。目的:淋巴管生成存在于肿瘤、炎症等许多疾病的病理生理过程中。本研究旨在探讨单核-巨噬细胞在炎症等病理环境中转分化成淋巴管内皮细胞的可能性。方法:取成人新鲜外周血,分离单核细胞,用内皮细胞培养基ECM培养,分别在FN铺板或VEGF-C、TNF-α、LPS、IL-3、IL-7刺激下诱导培养24 h或6 d,用RT-PCR和免疫细胞化学法,检测单核-巨噬细胞对淋巴管内皮细胞特异性标志物LYVE-1、Podoplanin、Prox1以及内皮细胞共同标志物vWF、eNOS的基因和蛋白表达。结果:单核-巨噬细胞在未诱导组,LYVE-1呈阳性表达,Podoplanin、Prox1、vWF、eNOS均表达阴性;经FN铺板或VEGF-C、TNF-α、LPS、IL-3、IL-7诱导24h或6d后,单核-巨噬细胞Podoplanin、Prox-1表达阳性,vWF和eNOS表达仍呈阴性。结论:FN、VEGF-C、TNF-α、LPS、IL-3、IL-7均能有效刺激单核-巨噬细胞表达淋巴管内皮细胞标志物。
[Abstract]:Background: lymphangiogenesis usually occurs in the pathophysiological processes of many diseases, such as tumor, trauma, inflammation and so on. Previous studies have shown that there are two main ways of lymphatic regeneration: the formation of new lymphatic vessels by budding of the original lymphatic vessels under the action of VEGF-C, and the migration of endothelial progenitor cells from peripheral blood to local tissues. After induction of VEGF-C, the cells were transformed into lymphatic endothelial cells (LECs). In tumor, trauma, inflammation and other pathological conditions, mononuclear cells migrate to the local tissue through blood circulation to convert into macrophages, which play an important role in lymphangiogenesis by producing VEGF-C. At present, it has been found that monocytes can differentiate into vascular endothelial cells under the action of VEGF, but whether they can be transformed into lymphatic endothelial cells induced by some factors has not been confirmed in vitro. In this study, cytokines produced by tissues in inflammatory and other pathological conditions were selected to activate monocytes. Such as fibronectin, lipopolysaccharide (LPSs) and tumor necrosis factor- 伪 (TNF- 伪), vascular endothelial growth factor (endothelial growm), interleukin-3interleukin-3 (IL-3), interleukin-7 (IL-7), et al. The expression of LYVE-1 Podoplanin Prox1 and endothelial markers vWF and Enos were detected, and the possibility of their transdifferentiation into lymphatic endothelial cells was observed, which provides a theoretical basis for further study on the mechanism of lymphatic angiogenesis. Objective: lymphangiogenesis exists in the pathophysiological processes of many diseases, such as tumor and inflammation. The aim of this study was to investigate the possibility of monocyte-macrophage transdifferentiation into lymphatic endothelial cells (LECs) in inflammatory and other pathological environments. Methods: monocytes were isolated from adult fresh peripheral blood and cultured on endothelial cell culture medium (ECM). Cultured 24 h or 6 d after stimulation with FN or VEGF-CnTNF- 伪 TPS- 伪 TNF- 伪 LPS- 伪 IL-3 IL-3 IL-7, RT-PCR and immunocytochemistry were used. The gene and protein expression of monocyte-macrophage specific marker LYVE-1 Podoplanin Prox1 and endothelial cell co-marker vWFFEnos were detected. Results: the positive expression of LYVE-1 in monocyte-macrophages was negative in the uninduced group, and the positive expression of VWF and Enos in monocyte-macrophages was still negative 24 or 6 days after being induced by FN or VEGF-CnTNF- 伪 -LPS-IL-3IL-7, and the positive expression of Podoplanintoprox-1 and Enos in monocyte-macrophages was still negative at 24 or 6 days after induction by FN or VEGF-CnTNF- 伪 -LPS-IL-3IL-7, the positive expression of Podoplanintoprox-1 in mononuclear macrophages was still negative. Conclusion the expression of lymphatic endothelial cell markers in mononuclear macrophages can be effectively stimulated by VEGF-Con TNF- 伪 and IL-3 / IL-7 in mononuclear macrophages.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R363

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