基于肝窦内皮细胞损伤评价黄芪来源成分的防护作用  

Evaluation the protective effects of the compounds from astragalus based on liver sinusoid endothelial cell injury

作　　者：赵志敏 黄恺 孙鑫[2] 沈丽[2] 刘成海[1,2,3] ZHAO Zhi-Min HUANG Kai SUN Xin LIU Cheng-Hai et al(Shanghai Key Laboratory of Traditional Chinese Clinical Medi- cine （ Shanghai ,201203 ） China)

机构地区：[1]上海市中医临床重点实验室,上海201203 [2]上海中医药大学附属曙光医院肝病研究所 [3]上海高校中医内科学E-研究院

出　　处：《中西医结合肝病杂志》2017年第3期155-157,I0004,I0005,共5页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：国家自然科学基金(No.81473404);中国博士后基金

摘　　要：目的:观察黄芪来源成分对氯化钴诱导SK-HEP-1细胞缺氧损伤的防护作用。方法:8001μM氯化钴作用于SK-HEP-1细胞24小时,诱导细胞缺氧损伤,不同浓度的药物成分作用于SK-HEP-1细胞,MTT法检测细胞活力,免疫荧光染色检测细胞vWF表达,高内涵图像分析系统观察细胞膜通透性、细胞核形态及溶酶体变化,荧光探针法检测NO、NOS水平。结果:与正常组比较,模型组细胞活力显著下降,vWF表达明显增加,细胞溶酶体平均荧光强度明显减弱,NO、NOS水平显著下降(P<0.05或P<0.01);药物处理后,细胞活力明显改善(P<0.01),其中黄芪甲苷和黄芪皂苷Ⅱ组vWF表达显著下降,溶酶体和NO水平显著提高(P<0.05);毛蕊异黄酮和毛蕊异黄酮-7-O-b-D-葡萄糖苷组vWF表达、溶酶体显著改善(P<0.05);芒柄花素和芒柄花素-7-O-b-D-葡萄糖苷组细胞NO、NOS水平显著提高,芒柄花素组细胞溶酶体荧光显著增强(P<0.05)。结论:黄芪来源的7种成分均有防护氯化钴诱导的SK-HEP-1细胞缺氧损伤的作用,但其作用基础存在细胞生物学差异。Objective：To observe the effects of the components from Astragalus on protecting SK-HEP-1 cells from hypoxia injury induced by cobaltous chloride （COC12）. Methods：SK-HEP-1 cell hypoxia injury was induced by 800μM CoC12 for 24 hours, and the cells were trea- ted with different concentrations of components. Cell viability was measured by MTr assay, vWF was detected by immunofluorescence stai- ning, the membrane permeability, nuclear morphology and lysosomal changes were observed by high content screen （HCS） system, NO and NOS were detected by fluorescence probe method. Results： Compared with the control group, 800μM CoCl2 decreased cell viability extreme- ly, the expression of vWF increased significantly, the mean fluorescence intensity of lysosomes, NO and NOS decreased significantly （ P 〈 0. 05 or P 〈 0.01 ）. After treatment, cell viability was significantly improved in different groups （P 〈 O. 01 ）. The expression of vWF was de- creased while lysosomes and NO levels increased significantly in Astragaloside Ⅳ and Astragaloside Ⅱ groups （ P 〈 0.05 ） ; in Calycosin and Genistein-7-O-b-D-glucoside groups, the expression of vWF was decreased but lysosome was improved significantly （ P 〈 0.05 ） ; NO and NOS levels in Formononetin and Ononin groups increased significantly, lysosomes in formononetin group were also significantly enhanced （P 〈 0.05 ）. Conclusion： The 7 compounds from astragalus have protective effects on SK-HEP-1 cells hypoxia injury induced by COC12, but there are some differences in cell biology.

关 键 词：黄芪来源成分 肝窦内皮细胞 缺氧损伤 防护作用 

分 类 号：R285[医药卫生—中药学]