调肝颗粒剂抗免疫性肝损伤的实验研究

1、调肝颗粒剂抗免疫性肝损伤的实验研究                 作者：陈忻， 赵晖， 张楠， 穆阳， 张秋云【关键词】  中药；中药药理；免疫性肝损伤；保肝摘要：目的观察调肝颗粒剂对卡介苗（BCG）和脂多糖（LPS）所致小鼠免疫性肝损伤模型的保护作用，并探讨其作用机制。方法BCG（1×104个活菌）静脉注射致敏小鼠，12 d后静脉给LPS（6g/只）攻击造成小鼠免疫性肝损伤，4 h后取血和肝脏，测血清AST和ALT并观察肝

2、脏病理变化；乙酰氨基酚腹腔注射（500 mg/kg）致肝损伤，2.5 h后处死小鼠测血清GSH和肝脏MDA。结果给BCG的同时给调肝颗粒剂（20 g/kg）能显着的减轻BCG和LPS所造成的免疫性肝损伤小鼠ALT，AST的升高，减轻肝细胞坏死，促进枯否氏细胞增生，促进肝细胞增生；并能显着减轻对乙酰氨基酚致肝损伤引起的GSH降低，同时减少肝脂质过氧化物生成。结论 调肝颗粒剂对BCG加LPS致小鼠免疫性肝损伤有显着的保护作用，这可能与它增加机体的抗氧化能力有关。关键词：中药；  中药药理；  免疫性肝损伤；  保肝Protective Effects of Tiaog

3、an Granules on Immunologic Liver Injury in MiceAbstract：ObjectiveTo investigate the protective effects of Tiaogan Granules on immunologic liver injury model. Methods(1) The liver injury model was induced by LPS intravenous injection in 12 days BCG interval, the mice were executed after 4h. the conte

4、nt of ALT and AST in serum were measured. (2) The other model was induced by paracetand ip 500 mg/kg, the mice were executed after 2.5h, the content of GSH and MDA in serum were measured. ResultsTiaogan granules were affused (dose20g/kg) for the mice when BCG were injected by intravenous at the same

5、 time. The increase of ALT and AST were reduced by intervention of Tiaogan granules and putrescence of hepatic cell were reduced too, but hepatic were hyperplasia and hepatic cells were improved. The decrease of GSH and the increase of MDA on the mice injected paracetamol were reduced by interventio

6、n of the Tiaogan granules.ConclusionThe protective effects of Tiaogan granules were confirmed on immunologic liver injury model. It was likely to be related to the enhancement of capability on resisted stress of oxidation of it.Key words：Traditional Chinese Medicine;  Parmacology of Traditional

7、 Chinese Medicine;  Immunologic Liver Injury;  Protective Effects on Liver Injury    调肝颗粒剂是肝病治疗专家、中华中医药学会内科肝病专业主任委员钱英教授沿用多年治疗乙型和甲、丙型肝炎的经验方。全方由槲寄生、黄芪等8味药组成，清热祛湿、疏肝利胆、补气养阴、活血通络。为阐述该方的保肝作用及作用机理，我们观察了调肝颗粒剂对小鼠免疫性肝损伤和化学性肝损伤的影响。现报道如下。1  材料与方法1  材料ICR雄性小鼠体重2530 g，由首都医科大学实验动物中心提

8、供。丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）检测试剂盒，北京中生北控生物科技股份有限公司产品。谷胱甘肽（GSH）、丙二醛（MDA）测定试剂盒为南京建成生物工程研究所产品。卡介苗（BCG）北京天坛生物制品研究所产品，批号20030704，脂多糖（LPS），美国Sigma公司产品。2  方法2.1  小鼠免疫性肝损伤模型制备参照文献1并做适当修改，ICR雄性小鼠2530 g，每鼠尾静脉注射BCG 0.2 ml（1×104个活菌）后正常饲养12 d，再经尾静脉注射LPS（6g/只）攻击，以诱导肝损伤，于4 h后眼眶取血，血浆离心（3 500 r/min

9、）10 min，血清测AST和ALT；同时取肝用10甲醛固定，制作石蜡切片做病理检查。2.2  乙酰氨基酚肝损伤模型制备参照文献2ICR小鼠腹腔注射乙酰氨基酚500 mg/kg，2.5 h后断颈放血处死，取肝脏测定GSH和MDA含量。2.3  给药方法调肝颗粒剂槲寄生、黄芪等8味药均为北京市药材公司市售饮片，制成100的水煎剂冰箱保存备用。ICR雄性小鼠25 30 g共24只，随机分为免疫性肝损伤模型组、损伤加调肝颗粒组和对照组，每组8只；调肝颗粒剂组于给BCG同时开始口服灌胃调肝颗粒剂100水煎剂20 g生药/kg，直至给LPS攻击当日，对照组同时给予生理盐水。另取ICR

10、小鼠雌雄各半，2025 g 共24只，随机分为乙酰氨基酚肝损伤组，损伤加调肝颗粒剂组和对照组，每组8只，调肝颗粒剂组给100调肝颗粒剂水煎剂20 g生药/kg。7 d后，给乙酰氨基酚造成肝损伤。对照组同时给予生理盐水。         1  徐叔云，卞如濂，陈  修. 药理实验方法学M.北京：人民卫生出版社，2003：1348.2  陈重阳，唐祖年，梁荣感.青蒿琥酯对正常及肝脏毒物中毒小鼠肝脏谷胱甘肽含量的影响J.中国药理学通报，1993，9（1）52.3  王根生，刘耕陶.一氧化氮在小鼠肝损伤中的作用J.中华医学杂志，1996，76（3）：203.4  王根生，刘耕陶. 枯否氏细胞与肝细胞损伤J.中国药理学通报，1994，10（5）：