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目的:研究五味子与淫羊藿混合提取物(SEE)对D-半乳糖诱导脑衰老小鼠学习记忆的影响,并探讨其相关机制。方法:将ICR小鼠随机分为5组,即空白组(蒸馏水灌胃,生理盐水皮下注射),衰老模型组(蒸馏水灌胃,D-半乳糖220 mg·kg~(-1)皮下注射),150,300,600 mg·kg~(-1)SEE组(SEE灌胃,D-半乳糖皮下注射)连续给药7周。通过避暗实验及跳台实验观察SEE对小鼠学习记忆能力的影响。应用水溶性四氮唑-1(water soluble tetrazolium-1,WST-1)法及硫代巴比妥酸法检测小鼠脑组织中超氧化物歧化酶(superoxide dismutase,SOD)活力与丙二醛(malonic dialdehyde,MDA)含量;通过实时荧光定量PCR检测小鼠脑组织p19,p53,p21 mRNA表达情况。通过亚油酸-硫氰酸铁法及Oyaizu法对SEE体外抗氧化能力进行检测。结果:避暗实验及跳台实验结果显示300,600 mg·kg~(-1)SEE可明显提高D-半乳糖诱导的脑衰老小鼠的学习记忆能力(P<0.05,P<0.01);同时,300,600 mg·kg~(-1)SEE组小鼠脑组织中SOD的活力明显增加(P<0.05,P<0.01),MDA含量明显减少(P<0.05,P<0.01),脑组织中p19,p53,p21 mRNA表达量明显降低(P<0.05,P<0.01)。亚油酸-硫氰酸铁法实验结果显示,反应体系建立36 h后,SEE对亚油酸的氧化抑制强于维生素E;Oyaizu实验结果显示,SEE对Fe3+的还原能力优于维生素E,吸光度达到0.711。结论:本研究明确了SEE能够改善D-半乳糖致脑衰老小鼠的学习记忆能力,该作用可能与其提高小鼠抗氧化能力以及降低小鼠脑组织中p19,p53,p21基因表达水平有关。
OBJECTIVE: To study the effect of mixed extract of Schisandra chinensis and Epimedium on the learning and memory of brain aging mice induced by D-galactose, and to explore its related mechanism. Methods: The ICR mice were randomly divided into 5 groups: blank group (distilled water, subcutaneous injection with normal saline), senescent model group (distilled water, intraperitoneal injection of D-galactose 220 mg · kg -1) , 150,300,600 mg · kg -1 SEE group (SEE intragastric administration, D-galactose subcutaneous injection) for 7 weeks. The effects of SEE on learning and memory ability of mice were observed by avoiding darkening experiment and jumping test. The activities of superoxide dismutase (SOD) and malondialdehyde (MDA) in brain tissue of mice were detected by water soluble tetrazolium-1 (WST-1) and thiobarbituric acid malonic dialdehyde (MDA)). The expression of p19, p53 and p21 mRNA in mouse brain were detected by real-time fluorescence quantitative PCR. The antioxidant capacity of SEE in vitro was tested by linoleic acid-ferric thiocyanate method and Oyaizu method. Results: The results of dark avoidance test and jumping test showed that SEE at 300 and 600 mg · kg -1 could significantly improve the learning and memory abilities in D-galactose-induced brain aging mice (P <0.05, P <0.01) (P <0.05, P <0.01). The levels of p19, p53 in the brain tissue of SEE group were significantly increased (P <0.05, P <0.01) , p21 mRNA expression was significantly decreased (P <0.05, P <0.01). Linoleic acid-ferric thiocyanate method showed that the oxidation of linoleic acid by SEE was stronger than that of vitamin E after 36 h reaction system was established. Oyaizu’s experimental results showed that the reducing ability of SEE to Fe3 + was better than that of vitamin E, and the absorbance Reached 0.711. CONCLUSIONS: This study clearly demonstrated that SEE can improve the learning and memory ability of brain-aged mice induced by D-galactose, which may be related to its ability to increase antioxidant capacity and reduce the expression of p19, p53 and p21 in mouse brain.