柿叶黄酮对肿瘤坏死因子α诱导大鼠血管平滑肌细胞凋亡信号调节激酶1表达的影响  

Effects of Flavone from Leaves of Diospyros kaki on Expression of Apoptosis Signal-Regulating Kinase 1 and Rat Vascular Smooth Muscle Cells Proliferation by Tumor Necrosis Factor α in Vitro

作　　者：欧阳平[1] 张彬[1] 贝伟剑[2] 赖文岩[1] 彭文烈[2] 许顶立[1] 

机构地区：[1]第一军医大学附属南方医院心血管内科,广东广州510515 [2]中山大学生命科学学院生物化学系,广东广州510275

出　　处：《中药材》2007年第7期819-822,共4页Journal of Chinese Medicinal Materials

基　　金：南方医科大学南方医院院长基金资助项目

摘　　要：目的:观察柿叶黄酮对肿瘤坏死因子α(TNF-α)诱导的大鼠血管平滑肌细胞凋亡信号调节激酶1的影响。方法:体外培养SD大鼠血管平滑肌细胞,应用柿叶黄酮与TNF-α共同进行作用并设立对照进行比较,采用非放射性的MTS/PES法确定血管平滑肌细胞的增殖状态。利用Western blotting蛋白免疫印迹法测定血管平滑肌细胞凋亡信号调节激酶1蛋白的表达情况。结果:各组的细胞增殖率分别为对照组0.817±0.074;TNF-α20 ng/ml组1.865±0.093;柿叶黄酮50μg/ml组0.905±0.044;TNF-α20 ng/ml联用柿叶黄酮50μg/ml组1.247±0.061。统计分析显示TNF-α20 ng/ml能明显刺激血管平滑肌细胞的增殖。柿叶黄酮单独作用对血管平滑肌细胞细胞增殖率无影响;柿叶黄酮对TNF-α诱导血管平滑肌细胞增于有明显的抑制作用,TNF-α对血管平滑肌细胞的凋亡信号调节激酶1蛋白的表达有显著的增强作用,这种作用部分可以被柿叶黄酮所抑制。结论:柿叶黄酮能明显抑制由TNF-α诱导的大鼠血管平滑肌细胞凋亡信号调节激酶1蛋白的表达。Objective ： To observe the effects of flavone from leaves of Diospyros kaki on expression of apoptosis signal-reguating kinase 1 （ASK1） and rat vascular smooth muscle cells （VSMCs） proliferation by tumor necrosis factor α in vitro. Methods： Rat aortic VSMCs were cultured in vitro and treated with tumor necrosis factor α（TNF-α） and flavone from leaves of Diospyros kaki, respectively, and were observed in comparison with the control group. The ratio of cell proliferation was determined by non-radioactive MTS/PES assay. The expression of ASK1 protein was evaluated by the immunoblotting technique using anti-ASK1 antibody. Results： The ratio of cell proliferation was 0. 817±0. 074 in the control group, and was 1. 865 ±0. 093 in TNF-α20 ng/ml group. It was shown that TNF- α　significantly induced rat VSMCs proliferation（ P 〈 0. 05 ）. The ratio of cell proliferatioh was 0. 905 ± 0. 044 in flavone from leaves of Diospyros kaki group corresponding to concentration of 50 μg/ml. It was shown that flavone from leaves of Diospyros kaki alone had no effect on rat VSMCs proliferation （p 〉 0. 05 ）. With TNF-α stimulation, flavone from leaves of Diospyros kaki significantly inhibited rat VSMCs proliferation （ 1. 247 ± 0. 061 vs 1. 865 ±0. 093, p 〈 0. 05 ）. The expression of ASK1 protein enhanced by TNF-α was significantly inhibited by flavone from leaves of Diospyros kaki in VSMCs. Conclusion ： Flavone from leaves of Diospyros kaki can significantly inhibit expression of ASK1 protein stimulated by TNF-α of rat VSMcs in vitro.

关 键 词：血管平滑肌细胞 柿叶 黄酮 细胞增殖 凋亡信号调节激酶1 

分 类 号：R285.5[医药卫生—中药学]