大鼠原代培养肝细胞免疫性损伤的病理特征及甘利欣的护肝作用

摘要

目的:探讨大鼠原代培养肝细胞免疫性损伤模型病理特征,评价甘利欣(Grz)护肝作用.方法:用卡介苗(BCG)整体致敏,脂多糖(LPS)离体攻击(BCG+LPS)致肝细胞损伤.常规测AST,LDH活性,Griess法测N0含量;免疫细胞化学法测ICAM-1表达,丫啶橙荧光染色法和流式细胞仪测肝细胞凋亡率,评价Grz护肝作用.结果:LPS对上清AST无显著影响.BCG+LPS使上清AST,LDH,N0,肝细胞ICAM-1表达及凋亡率均明显高于正常对照组;Grz抑制AST和LDH的同时,使NO和12 h肝细胞凋亡率显著降低.氨基胍对NO也有显著抑制.结论:ICAM-1高表达,大量N0生成和肝细胞凋亡可能是BCG+LPS诱发大鼠原代肝细胞免疫性损伤的病理现象;Grz可显著抑制NO生成和肝细胞凋亡.%AIM: To explore the pathologic characteristics of immunologic injury in primary cultured rat hepatocytes, and toevaluate the protective effect of glycyrrhizin (Grz) in vitro. METHODS: Sprague-Dawley rats were initiated withBCG vaccine in vivo, and then the primary cultured rat hepatocytes were stimulated with lipopolysaccharides(LPS) 10 mg/L in vitro (BCG+LPS treatment), the hepatoprotection by Grz was evaluated in the hepatocytestreated as above. Supernatant AST and LDH activities were measured, and nitric oxide (NO) was evaluated byGiress reaction. Aminoguanidine was used to confirm the target of Grz action. Expression of intercellular adhesionmolecular-1 (ICAM-1) was determined by immunocytochemistry. Percentage of apoptosis was detected withfluorescence microscope and flow cytometer. RESULTS: In LPS-treated group, supernatant AST was not in-creased compared with that in control, while supernatant AST and LDH in BCG+LPS treatment were increasedsignificantly (P＜0.05). The enhancements of AST and LDH were inhibited by co-culture with Grz (P＜0.05). Bothsupernatant NO and ICAM- 1 expression, and percentage of apoptosis in hepatocytes were elevated by BCG+LPStreatment (P＜0.01), and these elevations could be decreased by Grz co-incubation either (P＜0.05). The NOgeneration could be decreased by aminoguarnidine treatment (P＜0.05). CONCLUSION: The elevations of super-natant NO, ICAM-1 expression, and apoptosis in hepatocytes could be taken as three important pathologic changesin the immunologic hepatotoxicity induced by BCG+LPS treatment in primary cultured rat hepatocytes. NO syn-thesis mechanism was involved in the immunologic hepatotoxic process. Grz could downregulate both supernatantNO and hepatocyte apoptosis in the culture system.