發(fā)布時間：2018-06-14 06:56

  本文選題：單核細胞 + 轉(zhuǎn)分化��； 參考：《山東大學》2009年碩士論文

【摘要】： 研究背景:淋巴管新生通常發(fā)生在腫瘤、創(chuàng)傷、炎癥等許多疾病的病理生理過程中。以往的研究認為,淋巴管新生的途徑主要有二: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;外周血內(nèi)皮祖細胞遷移到局部組織,在VEGF-C的誘導下轉(zhuǎn)分化為淋巴管內(nèi)皮細胞。在腫瘤、創(chuàng)傷、炎癥等病理情況下,單核細胞通過血液循環(huán)遷移至局部組織轉(zhuǎn)化為巨噬細胞,通過產(chǎn)生VEGF-C,對淋巴管新生發(fā)揮重要作用。目前研究已發(fā)現(xiàn),單核細胞在VEGF等的作用下能夠向血管內(nèi)皮細胞轉(zhuǎn)分化,但能否在某些因子的誘導下轉(zhuǎn)分化為淋巴管內(nèi)皮細胞,尚未見體外研究證實。本研究選擇在炎癥等病理狀態(tài)下組織產(chǎn)生的能活化單核細胞的因子,如:細胞外基質(zhì)成分纖維連接蛋白(fibronectin,FN)、炎性因子脂多糖(lipopolysaccharide,LPS)和腫瘤壞死因子-α(tumor necrosis factor-α,TNF-α)以及血管內(nèi)皮生長因子C(vascular endothelial growm factor-C,VEGF-C)、白細胞介素-3(interleukin-3,IL-3)、白細胞介素-7(interleukin-7,IL-7)等,體外誘導單核細胞,檢測其淋巴管內(nèi)皮細胞標志物LYVE-1、Podoplanin、Prox1及內(nèi)皮細胞標志物vWF和eNOS的表達,觀察其向淋巴管內(nèi)皮細胞轉(zhuǎn)分化的可能性,為進一步研究淋巴管新生機理提供理論依據(jù)。 目的:淋巴管生成存在于腫瘤、炎癥等許多疾病的病理生理過程中。本研究旨在探討單核-巨噬細胞在炎癥等病理環(huán)境中轉(zhuǎn)分化成淋巴管內(nèi)皮細胞的可能性。 方法:取成人新鮮外周血,分離單核細胞,用內(nèi)皮細胞培養(yǎng)基ECM培養(yǎng),分別在FN鋪板或VEGF-C、TNF-α、LPS、IL-3、IL-7刺激下誘導培養(yǎng)24 h或6 d,用RT-PCR和免疫細胞化學法,檢測單核-巨噬細胞對淋巴管內(nèi)皮細胞特異性標志物LYVE-1、Podoplanin、Prox1以及內(nèi)皮細胞共同標志物vWF、eNOS的基因和蛋白表達。 結(jié)果:單核-巨噬細胞在未誘導組,LYVE-1呈陽性表達,Podoplanin、Prox1、vWF、eNOS均表達陰性;經(jīng)FN鋪板或VEGF-C、TNF-α、LPS、IL-3、IL-7誘導24h或6d后,單核-巨噬細胞Podoplanin、Prox-1表達陽性,vWF和eNOS表達仍呈陰性。 結(jié)論:FN、VEGF-C、TNF-α、LPS、IL-3、IL-7均能有效刺激單核-巨噬細胞表達淋巴管內(nèi)皮細胞標志物。
[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- 偽 (TNF- 偽), 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- 偽 TPS- 偽 TNF- 偽 LPS- 偽 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- 偽 -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- 偽 -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- 偽 and IL-3 / IL-7 in mononuclear macrophages.
【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R363

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本文編號：2016554