补肾宁心方对单核-内皮细胞在动脉粥样硬化中相互作用的影响

Effects of Bu-shen-ning-xin recipe on interaction of endothelial cells to monocytes in atherosclerosis

目的 探讨补肾宁心方对动脉粥样硬化中单核 -内皮细胞相互作用的影响。方法 以培养人脐静脉内皮细胞 (HUVECs)作为靶细胞 ,在内皮细胞培养基中加入氧化型低密度脂蛋白 (ox- LDL)并在试验体系中加入灌服补肾宁心方的兔血清 ,收集内皮细胞条件培养基 ,作用于人单核细胞系 U937细胞 ,检测内皮细胞条件培养基中 MCP- 1的含量及内皮细胞表面粘附分子、细胞间粘附分子 - 1 (ICAM- 1 )、血管细胞粘附分子 - 1(VCAM- 1 )的表达 ,同时测定 U937细胞 CCR2、LFA- 1以及 VL A- 4 m RNA水平。结果 ox- LDL显著升高内皮细胞条件培养基中 MCP- 1的含量 ,明显促进内皮细胞表面 ICAM- 1、VCAM- 1的表达 ,且在内皮细胞条件培养基作用下单核 U937细胞 CCR2、L FA- 1以及 VL A- 4 m RNA水平明显升高。如在试验体系中加入灌服补肾宁心方的动物血清 ,则 MCP- 1含量明显下降 ,内皮细胞表面 ICAM- 1、VCAM- 1的表达显著下调 (P<0 .0 1 ) ,且在条件培养基作用下单核 U937细胞 CCR2、LFA- 1以及 VLA- 4 m RNA水平明显下调。结论 补肾宁心方含药血清可能通过抑制内皮细胞分泌趋化因子、下调内皮细胞表面粘附分子的表达 ,抑制内皮细胞对单核细胞的趋化及相互粘附 ,从而发挥对血管内皮细胞的保护作用。

Objective To investigate effective mechanisms of the Bu-shen-ning-xin recipe on interaction of endothelial cells and monocytes in human atherosclerosis. Methods Using the cultured human umbilical vein endothelial cells (HUVECs) as target cells, the oxidized low density lipoprotein (ox-LDL) was added to the endothelial cell culture to prepare the injury model of human endothelial cell. The sera of rabbits having been afforded with the Bu-shen-ning-xin decoction was added to the experimental architecture, the conditioned medium of HUVECs (EC-CM) was collected and was used to act on U937(monocyte-like cell line). The content of monocyte chemoattractant protein(MCP-1) secreted by HUVECs was determined and the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) was measured. CCR2、LFA-1 and VLA-4 mRNA transcription of U937 were detected at the same time. Results Treatment of HUVECs with ox-LDL significantly increased the level of MCP-1 secreted by HUVECs and induced the expression of ICAM-1 and VCAM-1 of HUVECs. Under the action of EC-CM the mRNA expression of CCR2、LFA-1 and VLA-4 of U937 increased significantly. If serum of the animal having Bu-shen-ning-xin decoction was added to the experimental architecture, the level of MCP-1 decreased and the expression of ICAM-1 and VCAM-1 of HUVECs reduced significantly( P <0.01). The mRNA expressions of CCR2、LFA-1 and VLA-4 of U937 decreased significantly if treated with the EC-CM after having been incubated in sera of the animal having Bu-shen-ning-xin decoction . Conclusions The sera of the animal having Bu-shen-ning-xin decoction appears to have the protective effect on the vascular endothelial cell probably by the way of inhibiting the secretion of MCP-1, down-regulating the expression of ICAM-1 and VCAM-1 of endothelial cells and repressing the recruitment and adherence of monocytes to the endothelial cells.