抗菌药物对LPS诱导鸡肝细胞损伤的影响及复方甘草酸单胺的修复效应  

Effects of Antibiotics on LPS Induced Liver Cell Injury and Hepatoprotective Activity of Compound Ammonium Glycyrrhizin

作　　者：刘腾飞[1] 田静[1] 耿智霞[1] 李会敏[1] 余祖功[1] 

机构地区：[1]南京农业大学动物医学院,南京210095

出　　处：《畜牧兽医学报》2015年第2期309-316,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

摘　　要：旨在探索LPS诱导鸡肝细胞损伤过程中抗菌药物的影响及复方甘草酸单胺(CAG)的修复作用。选用半原位灌流法分离鸡肝细胞,原代培养48h,筛选LPS最佳致肝细胞损伤剂量;固定其最佳致损半量,并添加不同质量浓度的恩诺沙星、氟苯尼考、阿莫西林,观察致损效果;LPS(30μg·mL-1)+恩诺沙星(80μg·mL-1)致损24h,给予不同浓度CAG,检测细胞存活率及ALT、AST、SOD、GSH、MDA活性,流式细胞术检测细胞凋亡率。结果表明,LPS≤50μg·mL-1,肝细胞损伤不明显,LPS≥60μg·mL-1,肝细胞存活率极显著降低(P<0.01),ALT、AST活性极显著升高(P<0.01)。LPS(30μg·mL-1)各组随恩诺沙星、氟苯尼考、阿莫西林添加量增加,肝细胞损伤程度相应加大,LPS(30μg·mL-1)+恩诺沙星(80μg·mL-1)、LPS(30μg·mL-1)+氟苯尼考(40μg·mL-1)、LPS(30μg·mL-1)+阿莫西林(60μg·mL-1)可达肝细胞损伤标准。CAG剂量依赖性(25、50、100、200、400μg·mL-1)抑制LPS(30μg·mL-1)+恩诺沙星(80μg·mL-1)引起的ALT、AST、MDA活性和肝细胞早期凋亡比率的升高,提高SOD、GSH及肝细胞存活率。结果提示,LPS可致肝细胞损伤;抗菌药物使LPS更易诱发肝损伤;CAG通过抗氧化,并抑制细胞凋亡对鸡肝细胞发挥保护作用。The present study was aimed at the effects of antibiotics on LPS induced liver cell injury and hepatoprotective activity of compound ammonium glycyrrhizin(CAG).Chicken hepatocytes were isolated by semi-situperfusion method.After the cells were cultured for 48 h,LPS was added with different concentrations to screen the optimal dose of liver cell injury.At the half of the optimal dose of LPS,enrofloxacin,florfenicol and amoxicillin were added with different concentration to detect the indexes of liver cell injury.Hepatocytes were treated with different concentrations of CAG for 24 hafter the addition of LPS(30μg·mL-1)+enrofloxacin(80μg·mL-1).Cell viability and activities of ALT,AST,SOD,GSH,MDA were measured,flow cytometric analysis was conducted to detect the early apoptosis ratio of hepatocytes.The result showed that when LPS less than 50 μg· mL-1,the liver injury was not obvious.While LPS at more than 60μg·mL-1,the cell viability were significantly lower and the activities of ALT,AST in the culture media were significantly higher than those of controls(P<0.01).LPS(30μg·mL-1)combined with enrofloxacin(80μg·mL-1),florfenicol(40μg·mL-1),amoxicillin(60μg·mL-1)couldinduce liver cell injury significantly.CAG(25,50,100,200,400μg·mL-1)could markedly reduce the numbers of early apoptotic cells,inhibit the elevation of ALT,AST,MDA,and significantly increase the reduced level of SOD,GSH and cell viability.These results indicated that LPS could induce liver cell injury.Antibiotics might contribute to the liver cell injury induced by LPS.CAG counteracts liver cell injury at various levels by preventing apoptosis and oxidative stress damage.

关 键 词：LPS 恩诺沙星 氟苯尼考 阿莫西林 复合式肝细胞损伤 复方甘草酸单胺 

分 类 号：S859.7[农业科学—临床兽医学]