卡介苗联合脂多糖诱发大鼠原代培养肝细胞免疫性损伤模型的构建  

Establishment of BCG combined LPS-induced hepatocyte immunotoxicity model to assess liver protective effects

作　　者：郑芹珠[1] 王立明[1] 楼宜嘉[1] 

机构地区：[1]浙江大学药学院,浙江杭州310031

出　　处：《浙江大学学报（医学版）》2002年第6期419-423,456,共6页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金 (30 0 70 90 4 );浙江省医药卫生科研基金 (2 0 0 0 A0 4 9)资助项目

摘　　要：目的 :联合卡介苗 (BCG)和脂多糖 (L PS)构建大鼠原代培养肝细胞免疫性损伤模型 ,用于护肝药物的初步筛选。方法 :预注 BCG12 d的大鼠制备原代肝细胞 ,用 10 mg/ L L PS诱发肝细胞损伤。用联苯双酯 (DDB)、马洛替酯 (ML T)、水飞蓟宾 (SB)和甘利欣 (GRZ)进行防护 ,同法进行整体动物实验 ,以论证该模型的适用性和可靠性。测定血清和上清液中 AST、L DH和一氧化氮 (NO) ;计算肝脾指数 ;进行肝脏病理检查。结果 :体外实验中 ,BCG+L PS组上清液 AST、L DH和 NO明显高于正常对照组 (P<0 .0 1) ,且呈时间依赖性增加 ,12 h达稳态。DDB、ML T、SB和 GRZ均可明显抑制 12 h的 AST、L DH和 NO。整体实验中 ,BCG和 L PS处理 6 h后血清 AST、NO及肝脾指数明显高于正常对照组 (P<0 .0 1) ,L DH增加但无统计学差异 (P>0 .0 5 ) ;肝脏病理损伤明显。DDB、ML T、SB和GRZ对上述指标和病理损伤均有不同程度抑制作用。结论 :BCG联合 LObjective: To establish a hepatocyte immunotoxicity model for screening of liver protective medications. Methods: Cytotoxicity was induced by coincubating BCG pretreated rat hepatocytes in vivo and with 10 mg/L LPS in vitro . Biphenyldimethylesterate (DDB), malotilate(MLT), silybin(SB) and glycyrrhizin (GRZ) were coincubated along with LPS to prevent the hepatocyte injury and verify the applicability and reliability of the model . AST, LDH and nitric oxide (NO) were measured in both the serum and supernatant. The liver and spleen index were calculated and the liver histopathologic changes were examined microscopically. Results: Supernatant AST, LDH and NO in the BCG combined LPS group were increased in comparison with the control group ( P < 0.01 ). This increase was attenuated by the addition of DDB, MLT, SB and GRZ ( P <0.05). The serum AST, NO and liver and spleen index were also increased significantly compared with the control group ( P <0.01). Microscopic exam revealed serious histopathologic changes in the BCG combined LPS group. Hepatoxicity with associated liver enzyme elevation but histopathologic changes were attenuated by DDB, MLT, SB and GRZ. Conclusion: BCG combined LPS induced hepatocyte immunotoxicity in an in vitro rat model may be a useful technique to assess the effectiveness of liver protective medications.

关 键 词：卡介苗 副作用 脂多糖 肝疾病 药物疗法 疾病模型 原代肝细胞 一氧化氮 马洛替酯 水飞蓟宾 甘利欣 

分 类 号：R-332[医药卫生]