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目的:观察益气聪明汤及其拆方对氧化型低密度脂蛋白(ox-LDL)损伤血管内皮细胞(ECV-304)的保护作用并初探其作用机制。方法:以体外培养的人脐静脉内皮细胞株ECV-304为研究对象,以浓度为100μg/ml的ox-LDL制造氧化损伤细胞模型,然后分别加入益气聪明汤或其拆方的药物鼠血清进行培养,最后通过法检测细胞存活率、细胞裂解液中MDA含量、细胞上清中LDH活性,及检测调亡相关基因bcl-2、bax的mRNA表达来观察各药方对模型细胞上述各项指标变化的影响。结果:在100μg/ml的ox-LDL作用24小时后,内皮细胞存活率下降、细胞裂解液中MDA含量上升、细胞上清液中LDH活性上升,bcl-2mRNA表达下调而baxmRNA表达上调,内皮细胞增殖被明显抑制而其凋亡增加。益气聪明汤及其拆方的药物鼠血清能不同程度地使模型细胞LDH活性和MDA含量降低、bcl-2mRNA表达上调而baxmRNA表达下调,使得内皮细胞存活率由明显提高。结论:益气聪明汤及其拆方可通过程度不同地拮抗细胞氧化损伤,提高细胞生长活力并抑制其凋亡,发挥对血管内皮细胞的保护作用。
Objective: To observe the protective effect of Yiqi smart decoction and its disassembled prescriptions on oxidized low-density lipoprotein (ox-LDL)-damaged vascular endothelial cells (ECV-304) and to explore its mechanism of action. METHODS: Human umbilical vein endothelial cell line ECV-304 cultured in vitro was used as the research object. The oxidized damaged cell model was prepared with ox-LDL at a concentration of 100 μg/ml. After the culture, the survival rate of the cells, the MDA content in the cell lysate, the LDH activity in the cell supernatant, and the mRNA expression of the apoptosis-related genes bcl-2 and bax were measured by the method to observe the above-mentioned various indicators of the model cells. Impact of change. RESULTS: After exposure to 100 μg/ml ox-LDL for 24 hours, the viability of endothelial cells decreased, the MDA content in the cell lysates increased, the LDH activity in the cell supernatant increased, the expression of bcl-2 mRNA was down-regulated and the bax mRNA expression was up-regulated. Proliferation was significantly inhibited and its apoptosis increased. The Yiqi smart decoction and its deconstructed drug mouse serum can reduce the LDH activity and MDA content of the model cells, increase the expression of bcl-2 mRNA and downregulate the expression of bax mRNA in different degrees, making the survival rate of endothelial cells significantly improve. Conclusion: Yiqi smart decoction and its decomposing prescriptions can antagonize the oxidative damage of cells by different degrees, increase cell viability and inhibit its apoptosis, and exert its protective effect on vascular endothelial cells.