摘要
为研究黄芪总提物 (TEA)对肝细胞凋亡的保护作用及其机理 ,分别用D 氨基半乳糖 (D GalN ,70 0mg·kg- 1,ip) +脂多糖 (LPS ,1μg·kg- 1,ip)诱导小鼠在体肝细胞凋亡和H2 O2 (0 .1mmol·L- 1,1h)诱导原代培养大鼠肝细胞凋亡 ,采用形态学观察 ,DNA凝胶电泳和流式细胞术等方法检测细胞凋亡 .结果表明 :①TEA (40mg·kg- 1,ig× 2 ,5h)使D GalN +LPS升高的小鼠血中肿瘤坏死因子 (TNF)水平和肝脏丙二醛 (MDA)含量降低 ,以及使降低的肝线粒体锰 超氧化物歧化酶 (Mn SOD)活性升高 ;TEA可明显抑制D GalN +LPS引起的小鼠肝细胞皱缩变小 ,核染色质凝聚和DNA片段化 .②TEA (2 0mg·L- 1)可恢复或减轻由H2 O2 所致肝细胞增殖受抑和肝细胞MDA含量升高 ;TEA (40mg·L- 1)可使H2 O2 致DNA较强的AO荧光染色变淡 ,TEA(2 0mg·L- 1)对H2 O2 所致的DNA片段化有抑制作用 ,使H2 O2 升高的大鼠肝细胞DNA亚G1峰 (即凋亡峰 )明显降低 ,经DNA软件分析 ,TEA可使H2 O2 升高的细胞凋亡率从 6 3.7%降至 4 .2 % .提示 ,TEA对体内外肝细胞凋亡均有保护作用 。
To study the protective effect of total extract of Astragalus(TEA) against hepatocyte apoptosis and the mechanism of action, D galactosamine(D GalN, 700 mg·kg -1 , ip)+lipopolysaccharide(LPS, 1 μg·kg -1 , ip) induced hepatocyte apoptosis in mice and H 2O 2(0.1 mmol·L -1 , 1 h) induced apoptosis of primary cultured rat hepatocytes in vitro were made. Morphological assessment of apoptosis was performed with optical microscopy and fluorescent microscopy. DNA fragmentation was determined by flow cytometry and agarose gel electrophoresis of DNA. The results showed that the elevation of serum tumor necrosis factor level, malondialdehyde(MDA) content of liver homogenates and the decrease in Mn superoxide dismutase activity of hepatocytic mitochondrion in mice induced by D GalN+LPS were inhibited remarkably by TEA(40 mg·kg -1 , ig×2, 5 h) treatment. Hepatocyte shrinkage, chromatin condensation and DNA fragmentation from mice induced by D GalN+LPS were ameliorated remarkably by TEA. The inhibited proliferation and the elevation of MDA content of primary hepatocytes induced by H 2O 2 were reversed by TEA(20 mg·L -1 ) treatment. The chromatin condensation and DNA fragmentation of hepatocytes were inhibited by TEA(40 mg·L -1 ) treatment. The apoptosis peak level induced by H 2O 2 was reduced from 63.7% to 4.2% by TEA(20 mg·L -1 ). These results suggest that TEA has anti apoptosis effect on hepatocyte injury in vitro and in vivo. The action might be related to its anti oxidative activity.
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2001年第4期287-292,共6页
Chinese Journal of Pharmacology and Toxicology
基金
安徽省自然科学基金资助项目 (990 44 12 6 )
安徽省教委基金资助项目 (95LJ0 0 6 9)&&
