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目的:优选β-细辛醚,丁香酚和远志皂苷对Aβ25-35诱导的海马神经细胞的保护作用,确定三者最佳配比。方法:采用体外细胞培养的方法,通过NF染色鉴定,建立大鼠海马神经细胞Aβ25-35损伤模型。利用正交实验设计,测定细胞存活率(MTT法)、乳酸脱氢酶(LDH)活性,确定最佳三种药物最佳配比;通过观察细胞形态,利用Hoechst33258染色检测细胞凋亡情况。结果:L9(34)正交设计实验结果表明β-细辛醚(20,40,80μmol/L),丁香酚(2,4,8μmol/L),远志皂苷(10,20,40μmol/L)合用的最佳优化配比剂量分别为40μmol/L,2μmol/L,20μmol/L,可显著降低Aβ25-35(1μmol/L)诱导的神经细胞凋亡率(P<0.01)和LDH活性,提高细胞相对生存率。结论:三种药物联用可有效保护海马神经细胞免受β淀粉样蛋白的毒性损伤。
OBJECTIVE: To optimize the protective effect of β-asarone, eugenol and farnesin on hippocampal neurons induced by Aβ25-35, and to determine the optimal ratio of the three. Methods: Aβ25-35 injury model of rat hippocampal neurons was established by in vitro cell culture and NF staining. Orthogonal experimental design was used to determine the cell viability (MTT) and lactate dehydrogenase (LDH) activity to determine the optimal ratio of the three best drugs. Cell morphology was observed by Hoechst33258 staining. Results: The orthogonal experimental design of L9 (34) showed that β-asarone (20,40,80μmol / L), eugenol (2,4,8μmol / L) and teosaponin (10,20,40μmol / L) The optimal dosage of combination of 40μmol / L, 2μmol / L and 20μmol / L, respectively, could significantly reduce the apoptosis rate and the activity of LDH induced by Aβ25-35 (1μmol / L) Relative survival rate of cells. Conclusion: The combination of three drugs can effectively protect the hippocampal neurons from amyloid-β toxicity.