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目的探讨清心开窍方中皂苷、挥发油及多糖有效成分对淀粉样蛋白(Aβ)1-40海马注射诱导痴呆(Alzheimer’s disease,AD)大鼠海马区Bax、Bcl-2、Caspase-3及βAPP表达的影响。方法选取112只雄性SD大鼠,随机分为7组,每组16只,分别为正常组、假手术组、模型组、安理申组、皂苷组、挥发油组、多糖组。采用海马注射Aβ_(1-40)诱导AD大鼠模型。造模后第2天开始灌胃,正常组、假手术组、模型组给予等量双蒸水灌胃,安理申组[盐酸多奈哌齐片,1.67 mg/(kg·d)]、皂苷组[9 m L/(kg·d)]、挥发油组[3.33 m L/(kg·d)]、多糖组[8.33 m L/(kg·d)]灌胃,每天1次,连续2周(上午10:00)。灌胃结束后,采用Morris水迷宫测试大鼠的空间学习记忆能力;采用TUNEL染色检测海马CA1区细胞凋亡;采用免疫组化、实时定量荧光PCR及WB法检测AD大鼠海马区Bax、Bcl-2、Caspase-3及βAPP表达。结果造模前各组大鼠同一时间点的逃避潜伏期、穿台次数比较,差异均无统计学意义(P>0.05),且逃避潜伏期随时间推移逐渐缩短。造模后,与模型组比较,除挥发油组、多糖组外,安理申组、皂苷组逃避潜伏期均缩短,穿台次数显著增多(P<0.05,P<0.01);与模型组比较,皂苷组、挥发油组、多糖组大鼠海马CA1区凋亡细胞数量明显减少(P<0.05,P<0.01);Bcl-2表达上调,Bax、Caspase-3、βAPP表达下调,Bcl-2/Bax比值明显提高(P<0.05,P<0.01)。结论清心开窍方3种有效成分能不同程度地改善AD大鼠的学习记忆能力,其机制可能与降低海马区Bax、Caspase-3及βAPP表达,提高Bcl-2表达,抑制AD大鼠海马区内细胞凋亡有关。
Objective To investigate the effects of saponin, volatile oil and polysaccharide in Qingxin Kaiqiao Recipe on expressions of Bax, Bcl-2, Caspase-3 and βAPP in hippocampus of rats with amyloid (Aβ) 1-40 hippocampus injection-induced dementia influences. Methods One hundred and twelve male Sprague-Dawley rats were randomly divided into 7 groups (n = 16 each), which were normal group, sham operation group, model group, ambroxol group, saponin group, volatile oil group and polysaccharide group. AD rats were induced by injection of Aβ 1-40 into the hippocampus. The rats in the normal group, sham-operation group and model group were given the same amount of double-distilled water for gavage. Allixin group [donepezil hydrochloride tablets 1.67 mg / (kg · d)], saponin group [9 (3.33 m L / (kg · d)] and polysaccharide group [8.33 m L / (kg · d)] once daily for 2 weeks (morning 10 : 00). Morris water maze was used to test the spatial learning and memory abilities of rats. TUNEL staining was used to detect the apoptosis of hippocampal CA1 area. The expressions of Bax and Bcl-2 in hippocampus of AD rats were detected by immunohistochemistry, real-time quantitative PCR and WB method -2, Caspase-3 and βAPP expression. Results Before modeling, the escape latency and the number of times of wearing the same platform at the same time point in each group were not significantly different (P> 0.05), and the escape latency gradually shortened with time. Compared with the model group, the escape latency and the number of wearing trains of Amphenol and saponin groups were significantly increased (P <0.05, P <0.01) compared with the model group. Compared with the model group, the saponin group (P <0.05, P <0.01); the expression of Bcl-2, the expression of Bax, Caspase-3 and βAPP were down-regulated while the ratio of Bcl-2 / Bax was significantly higher in the volatile oil group and polysaccharide group Increase (P <0.05, P <0.01). Conclusion The three active ingredients of QXKT can ameliorate the learning and memory ability of AD rats to varying degrees. The mechanism may be related to the decrease of Bax, Caspase-3 and βAPP expression in hippocampus, the increase of Bcl-2 expression and the inhibition of AD rat hippocampus Apoptosis related.