【摘要】 目的检测切除小鼠荷瘤淋巴结后肺组织中与白细胞跨内皮迁移相关的趋化因子和黏附分子的表达,同时观察网状纤维改变以及波形蛋白(vimentin)及基质金属蛋白酶9(MMP9)表达情况,揭示它们在肺内预转移微环境形成中的变化及作用。方法将B16F10黑素瘤细胞接种至小鼠髂下淋巴结, 20只小鼠被平均分为手术切除荷瘤淋巴结和未切除荷瘤淋巴结组(对照组)。15 d后手术切除荷瘤淋巴结;切除3 d后,收集小鼠肺组织。应用镀银染色观察肺网状纤维变化;采用Western blot法检测CC趋化因子配体4(CCL4)、细胞间黏附分子1(ICAM-1)、血管细胞黏附分子1(VCAM-1)、 vimentin及MMP9的表达。结果与对照组相比,切除荷瘤淋巴结组肺组织内MMP9、 vimentin表达明显增高;网状纤维断裂明显,其面积百分比降低, CCL4、 ICAM-1、 VCAM-1表达均显著增加。结论切除小鼠荷瘤淋巴结促使肺组织内ICAM-1、 VCAM-1、 CCL4、 MMP9和vimentin的蛋白表达显著增加,有利于炎细胞黏附、跨越血管内皮进而形成炎性微环境。更多还原

【Abstract】 Objective To detect the expression of chemokine and adhesion molecules related to leukocytes’ transendothelial migration, meanwhile, to investigate changes of reticular fibers and the expression of vimentin and matrix metalloproteinase-9(MMP9) in lung tissues after surgical removal of mouse tumor-bearing lymph node, revealing their changes and roles in the formation of pre-metastatic microenvironment in the lung. Methods B16F10 melanoma cells were inoculated into mouse subiliac lymph node(SiLN). Twenty mice were equally divided into groups with or without(as a control group) tumor-bearing SiLN removal. Fifteen days later, tumor-bearing lymph node was surgically removed; 3 days after resection, mouse lung tissues were collected. The change of reticular fibers in lung tissues was observed by silver impregnation staining. The expression of C-C motif chemokine ligand 4(CCL4), intercellular adhesion molecule-1(ICAM-1), vascular cell adhesion molecule-1(VCAM-1), vimentin and MMP9 in lung tissues was detected by Western blotting. Results Compared with control group, expression of MMP9 and vimentin increased significantly in the lung tissues of SiLN removal group; reticular fibers were obviously fractured and its per area was reduced. Moreover, expression of CCL4, ICAM-1 and VCAM-1 also significantly increased. Conclusion Expression of CCL4, ICAM-1, VCAM-1, MMP9 and vimentin in mouse lung tissues is promoted after surgical removal of tumor-bearing lymph node, contributing to inflammatory cells’ adhesion to and extravasation across vascular endothelium and further resulting in the formation of inflammatory microenvironment.更多还原