基于均匀设计分析黄芪汤活性组分抗二甲基亚硝胺大鼠肝纤维化的配伍作用

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目的筛选古典方剂黄芪汤抗肝纤维化的有效组分及配伍作用。方法 (1)4周内给大鼠腹腔注射二甲基亚硝胺(dimethylnitrosamine,DMN)12次制备肝纤维化模型,于造模第3周首日开始给药干预至4周末。采用均匀设计对黄芪汤中4种组分(分别为黄芪总皂苷、黄芪总黄酮、甘草酸、甘草总黄酮)以4因素8水平安排给药;以肝组织羟脯氨酸(Hyp)含量和丙氨酸氨基转移酶(ALT)活性为观测指标,采用逐步回归法分析其抗肝纤维化效应的最优组方。(2)组分复方的再验证:模型制备及给药方法同上,采用上述实验获得的组分复方,与原方对照,观测大鼠肝组织胶原染色、Hyp含量及血清肝功能等变化。结果通过均匀设计研究,确定最优组方为组分复方(黄芪总皂苷、甘草酸的配伍比例为164:48),并得到有效验证。与模型组比较,组分复方及黄芪汤均可显著改善肝纤维化病理组织学变化,显著降低肝组织Hyp含量、血清ALT、天冬氨酸氨基转移酶(AST)以及碱性磷酸酶(ALP)活性(P<0.05);组分复方在降低肝组织Hyp含量及降低血清ALT活性方面均优于其分别单用组(P<0.05),降低血清ALT活性的作用优于黄芪汤原方(P<0.05)。结论黄芪总皂苷和甘草酸是黄芪汤发挥抗DMN大鼠肝纤维化作用的有效组分;两个组分配伍在降低肝组织胶原沉积及降低血清ALT活性方面具有明显的协同效应。 Objective To screen the effective components and compatibility of classical prescription Huangqi Decoction against hepatic fibrosis. Methods (1) Rats were intraperitoneally injected with dimethylnitrosamine (DMN) for 12 times to prepare the model of liver fibrosis within 4 weeks, and the intervention was started on the first day of the third week until the end of the fourth week. The four components (Astragalus total saponin, total flavonoids of Astragalus, glycyrrhizin, licorice total flavonoids) of Astragalus membranaceus in Astragalus membranaceus were evenly-designed and administered at the level of 4 factors and 8 respectively. The contents of hydroxyproline (Hyp) The activity of alanine aminotransferase (ALT) was taken as the observation index, and the stepwise regression method was used to analyze its optimal anti-hepatic fibrosis effect. (2) Re-validation of the components of the compound: The model was prepared and the method of administration was the same as above. The compound obtained by the above experiment was used for the comparison with the original one to observe the change of hepatic collagen staining, Hyp content and serum liver function. Results Through the uniform design and study, the optimal prescription was determined as the component compound (the ratio of total astragalosides and glycyrrhizic acid was 164: 48), which was validated. Compared with the model group, both compound and Huangqi decoction could significantly improve the histopathological changes of liver fibrosis, significantly reduce the Hyp content in liver tissue, serum ALT, AST and ALP ) Activity (P <0.05) .Compound compound was better than single use group (P <0.05) in reducing Hyp content in liver tissue and decreasing serum ALT activity, and the effect of lowering serum ALT activity was better than that of Astragalus soup (P < 0.05). Conclusion Astragalus saponin and glycyrrhizin are effective components of Astragalus membranaceus in inhibiting hepatic fibrosis in DMN rats. The combination of two components has obvious synergistic effect in reducing collagen deposition in liver tissue and decreasing serum ALT activity.
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