菖蒲四逆散干预AD模型大鼠Aβ代谢的级联靶效

目的 研究菖蒲四逆散对阿尔茨海默病(AD)模型大鼠学习记忆、海马 β 淀粉样蛋白前体(APP)、β-淀粉样蛋白(Aβ)1~42及血清超氧化物歧化酶(SOD)、丙二醛(MDA)、乙酰胆碱(Ach)、胆碱乙酰转移酶(ChAT)、乙酰胆碱酯酶(AChE)的影响,探讨菖蒲四逆散对AD模型大鼠Aβ代谢的干预及其与学习记忆能力、自由基氧化、神经递质改变的关联机制.方法 经水迷宫筛选合格的70只SPF级Wistar大鼠,随机选取10只为假手术组,其余大鼠脑立体定位注射Aβ25~35复制AD大鼠模型,以水迷宫学习记忆能力筛选造模成功的50只大鼠随机分为模型组、菖蒲四逆散低、中、高剂量组、阳性药组各10只.模型组与假手术组大鼠灌胃生理盐水2 ml/(100 g·d),菖蒲四逆散低、中、高剂量组及阳性药组大鼠分别灌胃对应药液2 ml/(100 g·d),连续28 d.给药25~28 d以Morris水迷宫测试各组大鼠学习记忆能力,然后取材检测海马APP、Aβ1~42及血清SOD、MDA、Ach、ChAT、AChE.结果 定位航行试验显示,与假手术组比较,模型组大鼠逃避潜伏期明显延长,在目标象限滞留时间较短;空间探索试验显示,首次到达原逃生平台位置的潜伏时间较长,穿越原平台位置及在目标象限滞留的时间明显较短,海马APP、Aβ1~42含量明显增高,血清SOD活性、ChAT活性、Ach水平明显降低,MDA水平、AchE活性明显增强(P<0.05,P<0.01).干预治疗后,与模型组比较,各给药组大鼠定位航行试验中逃避潜伏期均明显缩短,在目标象限滞留时间明显延长;空间探索试验中首次到达原平台位置潜伏期明显缩短,穿越原平台次数明显增多,海马APP、Aβ1~42明显降低血清SOD活性、ChAT活性、Ach水平明显提升,MDA水平、AchE活性明显降低(P<0.05,P<0.01).结论 菖蒲四逆散能有效改善AD模型大鼠学习记忆能力,其可能与通过干预AD模型大鼠海马Aβ代谢进而改善胆碱能神经递质、提升抗自由基氧化能力有关.

Cascade targeting effect on Changpu Sinisan intervening Aβmetabolism in Alzheimer's disease model rats

Objective To investigate effect of Changpu Sinisan on learning and memory abilities,Ach,ChAT,AChE,SOD,MDA in serum,APP,Aβ1~42 in hippocampus in Alzheimer's disease(AD)model rats,and explore the intervention of Changpu Sinisan on Aβmetabolism in AD model rats and its association with learning and memory ability,free radical oxidation and neurotransmitter changes.Methods 70 SPF Wistar rats were selected for learning and memory ability by water maze,10 rats were randomly selected(half female and half male)as sham group,the others were injected with Aβ25~35 by stereotatic techniques to copy AD model.50 rats were successfully divided into model,low,medium,high dose of Changpu Sinisan and positive drug groups,10 rats in each group.Rats in model and sham groups were treated with 2 ml/(100 g·d)normal saline by intragastric administration,rats in Changpu Sinisan and positive groups were treated with 2 ml/(100 g·d)corresponding drug by intragastric administration for 28 days.The learning and memory ability of rats in each group was tested by Morris water maze on the 25 th^28 th day of treatment,and the contents of Ach,ChAT,AChE,SOD,MDA in serum and APP,Aβ1~42 in hippocampus were determined.Results Compared with sham group,the escape latent period was significantly prolonged in place navigation experiment,the target quadrant was left less time,the latent time for the first time to reach the original escape platform was longer in spatial probe test,the residence time of crossing the original platform position and the target quadrant was shorter,the level of Ach,the activity of ChAT and SOD in serum were decreased;the level of MDA,the activity of AchE in serum were increased;the levels of APP,Aβ1~42 in hippocampus were increased in model group,the differences were statistically significant(P<0.05,P<0.01).Compared with model group,the escape latent period was shortened,the residence time of target quadrant was prolonged,the latent time for the first time to reach the original escape platform was shortened,the number of cross platform was increased,the level of Ach,the activity of ChAT and SOD in serum were increased,the level of MDA,the activity of AchE in serum were decreased,the levels of APP,Aβ1~42 in hippocampus were significantly decreased in treatment groups,the differences were statistically significant(P<0.05,P<0.01).Conclusions Changpu Sinisan could effectively improve the learning and memory of ability of AD model rats,which might be related to the improvement of cholinergic neurotransmitter,the promotion of against free radical oxidation capacity based on the intervention of Aβmetabolism.