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目的:通过均匀设计方法筛选出大川芎片有效组分保护缺氧损伤细胞最佳配伍比例,并找出相关通路对所得结果进行验证。方法:采用U10(108)均匀设计表,设置不同配伍组,以MTT法测定人神经母细胞瘤细胞SH-SY5Y存活率作为药效评价指标,将均匀设计方法与药效学实验相结合,经统计分析筛选出大川芎片有效组分保护缺氧损伤细胞最佳配比关系,同时设置验证实验来证明结果的可靠性。结果:Na2S2O4对诱导DSH-SY5Y细胞缺氧损伤有较好的效果,且各配伍组对缺氧损伤细胞均有不同程度的保护作用;CSZ均匀设计软件输出结果显示:当川芎多糖∶川芎酚酸∶川芎挥发油∶天麻多糖∶天麻总苷生药量配比为4.7∶1∶4.6∶8.7∶1.2时,保护缺氧损伤细胞效果最佳;通过通路图分析,明确乳酸脱氢酶LDH与琥珀酸脱氢酶SDH关系,可通过LDH结果验证MTT结果的可靠性。结论:复方按最佳配伍剂量入药可达到服用相同剂量使药效增强的效果,且组分入药成分清晰,为中药配伍的剂量关系及组分中药的研究探索出新的实验方法。
OBJECTIVE: To screen out the optimal proportion of Da Chuanxiong tablet effective components to protect hypoxic injury cells by uniform design method, and to find the relevant pathways to verify the results obtained. Methods: U10 (108) uniform design table, set up different compatibility groups, MTT assay SH-SY5Y survival rate of human neuroblastoma cells as a drug evaluation index, the uniform design method and pharmacodynamics experiments by the combination of Statistical analysis screened the effective components of Da Chuanxiong tablets to protect the optimal ratio of hypoxic injury cells, and set up verification experiments to prove the reliability of the results. Results: Na2S2O4 had a good effect on inducing hypoxia injury in DSH-SY5Y cells, and all compatibility groups had protective effects on hypoxic injury cells to some extent. The output of CSZ uniform design software showed that when polysaccharides of Chuanxiong rhizome: : Ligusticum chuanxiong volatile oil: Gastrodia elata polysaccharides: Gastrodia total glycosides crude drug dose ratio of 4.7:1:4.6:8.7:1.2, the best protective effect of hypoxic injury cells; through the pathway analysis, clear lactate dehydrogenase LDH and succinic acid off Hydrogenase SDH relationship, the reliability of MTT results can be verified by LDH results. Conclusion: The compound prescription can reach the effect of taking the same dose to enhance the efficacy, and the ingredients of the components are clear. It is a new experimental method to explore the dose-compatibility relationship of traditional Chinese medicine and components of traditional Chinese medicine.