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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 reconstruct Fufang Gannian prescription as Fufang Ganning granule, optimize its extraction process and evaluate its protective effect against carbon tetrachloride (CCl 4) -induced acute liver injury in mice. Methods: Optimize the formulation by Taguchi ethanol extraction process of Schisandra, Orthogonal test to optimize the water extraction process of the remaining herbs, the wet granulation process compound Gan Ning particles; 60 Kunming mice were randomly divided into 6 groups (normal CCl 4 model group, the bifendate positive group and the compound Ganning granule high, medium and low dose groups) were used to replicate the model of acute liver injury in mice with 0.1% CCl 4, and the levels of alanine aminotransferase ALT and AST activities, and the contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in the liver tissues. Results: The optimum alcohol extraction process was adding 0.5 times the amount of 75% ethanol reflux extraction 2 times, each 0.5 h; the best water extraction process plus 12 times the amount of water extracted three times, each time 1.5 h; Schisandra A and Salvia Sodium transfer rates were> 95%. Compared with the normal group, the activities of AST and ALT in the model group were significantly increased, while the activity of SOD in the liver tissue was decreased and the content of MDA was increased. Compared with the model group, Significantly reduce the activity of AST and ALT in serum of CCl 4 acute liver injury mice, reduce the content of MDA in liver homogenate and enhance the activity of SOD. Conclusion: The optimal extraction process is stable and feasible. Fufangganing Granule can significantly protect against acute liver injury induced by CCl_4 in mice by down-regulating the levels of ALT, AST and MDA and up-regulating the level of SOD.