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目的:观察SD大鼠海马立体定向注射β-淀粉样蛋白1-40后β位淀粉样前体蛋白裂解酶1mRNA表达的变化及加减地黄饮子对其的干预作用。方法:①实验于2005-09/2006-09在齐齐哈尔医学院医药科学研究所完成。②选用100只SD大鼠随机分为空白对照组、假手术对照组、模型对照组、盐酸多奈哌齐组、加减地黄饮子组共5组,每组20只。通过海马立体定向注射β-淀粉样蛋白1-40诱导老年性痴呆动物模型。③盐酸多奈哌齐按0.33mg/(kg·d)给药,加减地黄饮子组按1.0g/(kg·d)给药,共给药28d。空白对照组和假手术对照组给予等量生理盐水。④第5周处死大鼠,应用实时定量PCR法检测大脑海马组织β位淀粉样前体蛋白裂解酶1mRNA表达(用2-ΔΔCt表示,Ct为阈循环值,ΔCt=CtBACE1-CtGAPDH,ΔΔCt=ΔCt各干预组-ΔCt空白对照组)。⑤多组间差异的显著性分析用单因素方差分析,组间两两比较运用LSD-t检验。结果:实验选用100只大鼠,每组随机选5只用于抽提大脑海马组织总RNA,因此共有25只大鼠纳入结果分析。β位淀粉样前体蛋白裂解酶12-ΔΔCt值模型组(4.67±0.52)显著高于假手术对照组(1.07±0.08)(P<0.01),表明模型组β位淀粉样前体蛋白裂解酶1mRNA表达上调。盐酸多奈哌齐组(1.80±0.23)和加减地黄饮子组(1.26±0.20)显著低于模型对照组,表明β位淀粉样前体蛋白裂解酶1mRNA表达下调。结论:大鼠大脑海马注射β-淀粉样蛋白1-40后海马组织β位淀粉样前体蛋白裂解酶1mRNA表达水平明显增高,加减地黄饮子提取物可以抑制海马组织β位淀粉样前体蛋白裂解酶1mRNA的表达,从而发挥抗老年性痴呆的作用。
OBJECTIVE: To observe the changes of β-amyloid precursor protein lyase 1 mRNA expression in the hippocampus of SD rats after stereotactic injection of β-amyloid protein 1-40, and to observe the effects of diterpenes and decoction on it. METHODS: 1 The experiment was completed at the Institute of Medical Sciences, Qiqihar Medical College from September 2005 to September 2006. 2100 Sprague-Dawley rats were randomly divided into blank control group, sham-operated control group, model control group, Donepezil hydrochloride group, and Radix Rehmanniae subgroup. There were 5 groups, 20 in each group. Animal model of Alzheimer’s disease was induced by stereotactic injection of β-amyloid 1-40 in the hippocampus. 3 Donepezil hydrochloride was administered at 0.33mg/(kg·d), and rehmannia glutinosa was administered at 1.0g/(kg·d) for 28 days. The same amount of physiological saline was given to the blank control group and the sham control group. 4The rats were sacrificed at the 5th week and real-time quantitative PCR was used to detect the expression of β amyloid precursor protein lyase 1 mRNA in the hippocampus of the brain (represented by 2-ΔΔCt, Ct was the threshold cycle value, ΔCt=CtBACE1-CtGAPDH, and ΔΔCt=ΔCt. Each intervention group - ΔCt blank control group). The significance of the differences between the groups was analyzed by one-way analysis of variance (ANOVA), and the LSD-t test was used for comparison between groups. RESULTS: A total of 100 rats were selected for the experiment. Five rats in each group were randomly selected for the extraction of total hippocampal RNA. A total of 25 rats were included in the analysis of the results. The β-amyloid precursor protein lyase 12-ΔΔCt value in the model group (4.67±0.52) was significantly higher than the sham-operated control group (1.07±0.08) (P<0.01), indicating that the β-amyloid precursor protein lyase 1mRNA upregulation. Donepezil hydrochloride (1.80±0.23) and Radix Rehabilitation group (1.26±0.20) were significantly lower than the model control group, indicating that β-amyloid precursor protein lyase 1 mRNA expression was down-regulated. Conclusion: The expression level of β-amyloid precursor proteolytic enzyme-1 mRNA in the hippocampus of rat hippocampus after β-amyloid 1-40 injection is significantly increased. The addition and subtraction of Dihuangyinzi extract can inhibit the β-amyloid precursor in hippocampus. The expression of proteolytic enzyme 1 mRNA thus exerts the effect of anti-senile dementia.