关键词：
鸡原代肝细胞;内毒素;替米考星;肝损伤;凋亡
摘 要：
旨在探索内毒素(LPS)和替米考星联合诱导鸡肝细胞损伤的机制。选取400g左右海蓝公鸡,采用改良二步罐流法分离肝细胞。用噻唑盐比色法(MTT法)测定细胞存活率,改良赖氏法测定细胞培养液中谷丙转氨酶(ALT)、谷草转氨酶(AST)的含量并筛选出LPS、替米考星单独致鸡肝细胞损伤的最佳剂量和LPS与替米考星联合致鸡肝细胞损伤的最佳剂量,探讨此复合式肝损伤过程中的相互影响。Hoechest 33342对肝细胞染色、Caspase-3活性的测定和流式细胞仪来探究细胞是否出现凋亡。结果表明:LPS浓度为30μg/mL,替米考星浓度为120μg/mL时,肝细胞存活率极显著降低(P<0.01);检测上清中ALT、AST出现极显著差异(P<0.01),肝细胞损伤显著;细胞荧光照片可看出,凋亡细胞明显增多,Caspase-3活性明显升高,流式细胞仪检测细胞凋亡率明显升高。结果提示,内毒素和替米考星可单独诱发鸡肝细胞损伤并造成细胞凋亡,二者联合可能会使损伤作用增强。
译 名：
Chicken primary hepatocyte injury in vitro induced by lipopolysaccharides and tilmicosin
作 者：
WU Feng;CUI Jingyi;GUO Xunwen;AN Ran;YU Zugong;College of Veterinary Medicine,Nanjing Agricultural University;
关键词：
chicken primary hepatocyte;;lipopolysaccharides;;liver injury;;apoptosis;;tilmicosin
摘 要：
The aim of this study is to explore the mechanisms of chicken primary hepatocytes injury induced by lipopolysaccharides and tilmicosin. Hepatocytes were isolated from male Hailan chickens weighing 400 g by a two-step collagenase perfusion method. The cell viability was measured by MTT,and the alanine aminotransferase( ALT) and alanine transaminase( AST) by improved Lai method. Optimum concentration was selected by these methods to research this influence. Apoptosis was determined by Hoechest 33342 and activity of caspase-3. The results shows that the viability of hepatocytes decreased( P < 0. 01) when concentration of LPS was 30μg/mL and tilmicosin was120μg/mL; the ALT and AST was increased to 30. 58±4. 41 and 23. 57±2. 49,respectively( P<0. 01). According to the picture of fluorescence,the apoptotic cells and activity of caspase-3( 0. 353±0. 06) was increased.
