摘要
目的研究党参多糖对低浓度过氧化氢(H2O2)刺激下EA.hy926细胞的保护作用。方法采用体外培养EA.hy926细胞,利用H_2O_2小剂量长时间刺激进行造模,并选用维生素E作为阳性对照组。采用MTT法检测细胞的增殖情况,光学显微镜观察细胞形态学的改变,SA-β-半乳糖苷酶染色法检测细胞的衰老水平,流式细胞术检测细胞周期,比色法检测细胞细胞培养上清中一氧化氮含量及细胞内的超氧化物歧化酶、丙二醛的含量,ELISA检测细胞培养基中内皮素-1、诱导型一氧化氮合酶、内皮型一氧化氮合酶的水平,DCFH-DA检测细胞内活性氧的水平。结果党参多糖各浓度组、维生素E组与模型组比较细胞的形态均有不同程度改善。与模型组比较,党参多糖各浓度组、维生素E组细胞的存活率升高,细胞G1期比例下降,SA-β-半乳糖苷酶染色的细胞蓝染率下降,超氧化物歧化酶、一氧化氮、内皮型一氧化氮合酶的水平升高,丙二醛、诱导型一氧化氮合酶、内皮素-1、活性氧水平下降。结论党参多糖可能通过提高EA.hy926细胞的抗氧化能力、调控内皮素-1与一氧化氮的动态平衡发挥其对氧化损伤的保护作用。
Objective To study the protective effect of Codonopsis pilosula polysaccharides on EA.hy926 cells stimulated by low concentrations of hydrogen peroxide（H2O2）.Methods EA.hy926 cells were cultivated in vitro and H2O2 was used for chronic stimulation to establish a model before vitamin E was used as the positive control group.The MTT method was used to detect the inhibition rate of cell proliferation,cell morphology was observed under an optical microscope,SA-beta galactosidase staining method was used to detect cells for the state of senescence and cell cycles were detected by flow cytometry.In addition,colorimetry assay was used to detect the content of nitric oxide in the cell culture medium and the content of SOD and MDA in cells.Enzyme-linked immunosorbent assay was used to detect the cell culture medium for the levels of endothelin-1,inducible nitric oxide synthase and endothelial nitric oxide synthase while DCFH-DA fluorescent chromogenic method was used to determine the levels of intracellular reactive oxygen species.Results The cell morphology of Codonopsis pilosula polysaccharides in each concentration group and vitamin E group was improved to different degrees compared with the model group.Compared with the model group,the survival rate of cells was increased,the percentage of G1 phase was decreased,the rate of beta galactosidase staining of cells was decreased,SOD levels were increased and levels of MDA were decreased in all the other groups.The levels of i NOS and ET-1 were decreased,the levels of e NOS and NO were increased,and the level of ROS was decreased in those groups.Conclusion Codonopsis pilosula polysaccharides can protect the EA.hy926 cells from oxidative damage by enhancing the antioxidant activity and regulating the dynamic balance between ET-1 and NO.
作者
王慧云
李义波
谢荣丹
张帆
WANG Hui-yun;LI Yi-bo;XIE Rong-dan;ZHANG Fan(Key Laboratory of Molecular Medicine for Major Diseases and Prevention with Traditional Chinese Medicine of Gansu Province,Gansu University of Chinese Medicine,Lanzhou 730000,China)
出处
《解放军药学学报》
CAS
CSCD
2018年第3期195-199,204,共6页
Pharmaceutical Journal of Chinese People's Liberation Army
基金
甘肃省中医药管理局科研项目
No.GZK-2013-2
