摘要
目的:观察氧化低密度脂蛋白(ox-LDL)损伤的血管内皮细胞(HUVEC)与人白血病单核细胞(THP-1)的黏附作用,以及通络药物(通心络超微粉溶液和人参皂苷Rb1)干预的影响。方法:采用ox-LDL建立血管内皮细胞损伤模型,将细胞分为正常对照组、模型组、通心络组和人参皂苷Rb1组。实验采用MTS比色法检测ox-LDL损伤的HUVEC的生存活性,用活细胞染色方法观察不同组HUVEC与THP-1细胞的黏附率,用酶联免疫吸附测定(ELISA)方法检测HUVEC培养上清中单核细胞趋化因子(MCP-1)、可溶性血管内皮细胞间黏附分子(s VCAM-1)、可溶性内皮细胞间黏附分子(sICAM-1)、E-选择素(E-selectin)的水平,用蛋白免疫印迹法(Western Blot)检测各组HUVEC条件培养基培养的单核细胞趋化因子受体2(CCR2)、极迟抗原4(VLA-4)、巨噬细胞分化抗原-1(Mac-1)的表达。结果:与正常对照组(100.00±1.31)%比较,模型组HUVEC生存活性降低为正常对照组的(75.57±1.02)%,通心络组和人参皂苷Rb1组HUVEC生存活性明显分别提高了(99.25±1.40)%、(99.48±2.15)%。与正常对照组比较,模型组黏附于HUVEC的THP-1细胞数量明显增多,与模型组比较,通心络组和人参皂苷Rb1组黏附于HUVEC细胞的THP-1细胞数量减少。通心络组和人参皂苷Rb1组HUVEC细胞培养上清中MCP-1、sVCAM-1、sICAM-1、E-selectin的水平和THP-1细胞上相应受体CCR2、VLA-4、Mac-1的表达较模型组降低。上述比较差异均有统计学意义(P<0.05)。结论:通心络和人参皂苷Rb1能够保护HUVEC,减少ox-LDL损伤的血管内皮细胞趋化、黏附分子的分泌及单核细胞上相应受体的表达,从而抑制单核细胞向受损血管内皮的趋化黏附。
Objective： To observe the effect of oxidative-low density lipoprotein（ox-LDL） injured human leukemia mononuclear cells（THP-1） adhesion to human umbilical vein endothelial cells（HUVECs） in vitro with the intervening function of dredging collateral drug, tongxinluo（TXL） and ginsenoside（Rb1）.Methods： Cell injury was induced by ox-LDL treatment. The cells were divided into 4 groups：① Normal control group,② Injury model group, the cells were cultured by ox-LDL,③ TXL group, the cells were cultured with both ox-LDL and TXL,（4） Rb1 group. HUVEC viability was measured by MTS assay, adherence rate of THP-1 cells to HUVECs was tested by vital cell staining. The contents of monocyte chemoat-tractant protein（MCP-1）, soluble vascular cell adhesion molecule（sV CAM-1）, soluble inter vascular cell adhesion molecule-1（sICAM-1） and E-selectin in HUVEC conditioned medium were detected by ELISA; protein expressions of CCR2, VLA4 and Mac-1 in THP-1 cells were examined by Western blot analysis.Results： Compared with Control group, HUVEC viability was decreased in Injury model group（100 ±1.31） % vs（75.57 ± 1.02） %, while increased in both TXL and Rb1 groups（99.25 ± 1.40） % and（99.48 ± 2.15） %; Injury model group showed elevated adherence rate of THP-1 cells to HUVECs, while the adherence rates were reduced in both TXL and Rb1 groups. Compared with Injury model group, TXL group and Rb1 group showed decreased levels of MCP-1, sV CAM-1, sI CAM-1 and E-selectin in HUVEC conditioned medium; decreased protein expressions of CCR2, VLA4 and Mac-1 in THP-1 cells.Conclusion： TXL and Rb1 could protect HUVECs, reduce ox-LDL injury induced vascular endothelial cell adhesion and decrease relevant receptor expression in monocytes; therefore, inhibit injured monocytes adherence to vascular endothelial cells.
出处
《中国循环杂志》
CSCD
北大核心
2016年第5期480-483,共4页
Chinese Circulation Journal
基金
国家重点基础研究发展计划973计划(2012CB518606)
