黄芪多糖对细胞增殖及血管内皮细胞与白细胞粘附作用的影响

Effects of astragalus polysacharin on fibroblast proliferation and adhesion between HUVECs and white cells

目的观察黄芪多糖对成纤维细胞与血管内皮细胞增殖效应及对血管内皮细胞与白细胞粘附作用的影响。方法采用MTT法观察体外培养的人正常成纤维细胞和慢性创面的创缘成纤维细胞的增殖和人脐静脉血管内皮细胞(HUVEC)增殖;采用虎红法、荧光免疫组化法在TNF刺激的HUVEC模型上观察黄芪多糖对人外周血中性粒细胞(PMN)、单核细胞TPH-1与HUVEC粘附及HUVEC表面粘附分子的表达。结果在244-156mg/L范围内,黄芪多糖作用下创缘成纤维细胞增殖率明显高于对照组(P<001);244-39mg/L黄芪多糖作用下人正常皮肤的成纤维细胞增殖率明显高于对照组(分别P<001,P<005);在975-156mg/L浓度范围内对HUVEC未表现出明显的增殖作用。在4h时25mg/L黄芪多糖组PMN与HUVEC的粘附量明显低于TNF组(P<005);25mg/L和100mg/L黄芪多糖组TPH-1与HUVEC的粘附量明显低于TNF组(分别P<001和P<005);作用12h时,25-100mg/L黄芪多糖组HUVEC与PMN粘附量明显低于TNF组(分别P<005,P<001和P<001);50mg/L和100mg/L也使TPH-1与HUVEC粘附量明显低于TNF组(分别P<001和P<005)。与TNF组比较,在25-100mg/L黄芪多糖作用4h能明显降低VCAM-1、ICAM-1的荧光强度(分别P<001和P<005)。作用12h,与TNF组比,50mg/L黄芪多糖组CD44的荧光强度明显低于TNF组(P<005)。

AIM: To investigate the effects of Astragalus polysacharin(APS) on human fibroblast and human umbilical vein endothelia cell (HUVEC) proliferation, as well as its acts on adhesion between white cells and HUVECs. METHODS: Human fibroblasts from distal and proximal skin away the ulcer were cultured as normal fibroblasts(NF) and wounded fibroblasts(WF). MTT assay was used for detecting cell proliferation, Rose Bengal staining and fluorescence immunohistology assay were used for examining the adhesion of human polymorpho-nuclear cell(PMN) and TPH-1 to HUVECs. RESULTS: 2 44-156 mg/L APS promoted WF proliferation, and 2 44-39 mg/L APS also promoted NF proliferation, but it did not show any proliferating effect on HUVECs. APS inhibited the adhesion of PMN or TPH-1 to HUVECs induced by tumor necrosis factor(TNF). At 25-100 mg/L, it also inhibited both VCAM-1 and ICAM-1 expression in HUVECs induced by TNF. Treatment with APS for 12 h also inhibited CD44 expression in HUVECs. CONCLUSION: APS shows mitogenic activity on both human normal and wounded fibroblasts. It also exerts anti-inflammation effects by inhibiting adhesion molecule expression and adhesion of white cells to HUVECs. [