复方肝宁颗粒的制备及其对CCl_4诱导小鼠急性肝损伤的保护作用

Preparation of Compound Ganning Granules and Investigation of Its Protective Effect on Acute Hepatic Injury Induced by CCl_4 in Mice

目的:将复方肝宁经验方改制成复方肝宁颗粒,优化其提取工艺并评价该制剂对四氯化碳(CCl_4)诱导的小鼠急性肝损伤的保护作用。方法:通过Taguchi设计优化处方中五味子的乙醇提取工艺,正交试验优化其余药材的水提工艺,采用湿法制粒工艺得到复方肝宁颗粒;将60只昆明种小鼠,随机分成6组(正常组,CCl_4模型组,联苯双酯阳性组和复方肝宁颗粒高、中、低剂量组),用0.1%CCl_4复制小鼠急性肝损伤模型,测定小鼠血清中丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)活性,肝组织中超氧化物歧化酶(SOD)和丙二醛(MDA)的含量。结果:最佳醇提工艺为加12.5倍量75%乙醇回流提取2次,每次0.5 h;最佳水提工艺为加12倍量水提取3次,每次1.5 h;五味子醇甲及丹参素钠转移率均>95%。与正常组比较,模型组小鼠血清中AST和ALT的活性明显升高,肝组织中SOD活性降低,MDA含量升高;与模型组比较,复方肝宁颗粒高、中、低剂量组均能显著降低CCl_4急性肝损伤小鼠血清中AST和ALT的活性,降低肝匀浆中MDA的含量,增强SOD的活性。结论:优选的提取工艺稳定可行。复方肝宁颗粒通过下调ALT,AST和MDA水平,上调SOD水平,对CCl_4造成的小鼠急性肝损伤具有显著的保护作用。

Objective： To prepare compound Ganning granules, optimize its extraction process and investigate its protective effect on acute hepatic injury induced by CC14 in mice. Method： Ethanol extraction process of Schisandrae Chinensis Fructus was optimized by Taguchi design, water extraction process of the other herbs was optimized by orthogonal test. Compound Ganning granules was prepared through wet granulation process. Mice were randomly divided into six groups, included normal group, model group, bifendate group and high, middle, low dose group of compound Ganning granules. Acute hepatic injury mice model was induced by 0.1% CC14. Levels of alanine aminotransferase （ALT）, aspartate aminotransferase （AST） in serum and malonialdehyde （MDA）, superoxide dismutase （SOD） in live tissue were detected. Result： After being optimized, transfer rates of schisandrin A and sodium danshensu were more than 90%. Compared with the normal group, activities of ALT, AST and level of MDA in model group were significantly increased and activity of SOD was inhibited. Compared with the model group, activities of ALT, AST and level of MDA in compound Ganning granules groups were significantly decreased and activity of SOD was increased. Conclusion： These optimized processes are stable and feasible. Compound Ganning granules has hepatoprotective effect on acute hepatic injury induced by CC14 in mice by up-regulation of activity of SOD and down-regulation of activities of ALT, AST and MDA.