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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ECM培养,分别在FN铺板或VEGF-C、TNF-α、LPS刺激下诱导培养24 h,用RT-PCR和免疫荧光细胞化学方法检测单核-巨噬细胞对淋巴管内皮特异性标志物LYVE-1、Podoplanin、Prox1及内皮细胞共同标志物vWF、eNOS的基因和蛋白表达。结果在未诱导组,单核-巨噬细胞LYVE-1呈阳性表达,Podoplanin、Prox1、vWF、eNOS表达呈阴性;在FN铺板或VEGF-C、TNF-α、LPS诱导组,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均能有效刺激单核-巨噬细胞表达淋巴管内皮标志物。
Purpose Lymphangiogenesis exists in the pathophysiology of many diseases such as cancer and inflammation. This study aimed to investigate the possibility of monocyte-macrophage transdifferentiation into lymphatic endothelial cells in an inflammatory environment. Methods Monocytes were isolated from fresh adult human peripheral blood and cultured in ECM. The cells were cultured for 24 h under the stimulation of FN or VEGF-C, TNF-α and LPS, respectively. The mononuclear cells were detected by RT-PCR and immunofluorescence cytochemistry The gene and protein expression of macrophages on lymphatic endothelial specific markers LYVE-1, Podoplanin, Prox1 and endothelial marker vWF, eNOS. Results The positive expression of LYVE-1 in mononuclear macrophages and the expression of Podoplanin, Prox1, vWF and eNOS in non-induced group were negative. In FN-plated or VEGF-C group, TNF- Podoplanin, a positive expression of Prox-1, and a negative expression of vWF and eNOS. Conclusion FN, VEGF-C, TNF-α and LPS can effectively stimulate the expression of lymphatic endothelial markers in monocytes-macrophages.