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,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13位淀粉样前体蛋白裂解酶1mRNA表达(用2^-△△ct表示,Ct为阈循环值,△Ct=Ctbace1CtGAPDH,△△Ct=△Ct各干预组-△Ct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-f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^-△△ct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B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13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,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。
AIM: To observe the effect of modified decoction of rehmanniae on β-site amyloid precursor protein (APP) cleaving enzyme (BACE1) mRNA expression in hippocampus tissue of SD rats induced by 13-amyloid 1-40 injection into brain.METHODS:①This study was completed at the Medicine Institute of Qiqihar Medical College from September 2005 to September 2006.②Totally 100 SD rats were enrolled and randomly divided into 5 groups with 20 in each, named as blank controls group, sham controls group, model group, Donepezil group and modified decoction of rehmanniae group. Rats were induced by 13-amyloid1-40 sterotaxis injection into the bilateral hippocampus for Alzheimer's disease (AD) model. ③Physiological saline was injected into the rats of blank controls group and sham controls group. And 0.33 mg/kg Donepezil was administrated for Donepezil group, while 1.0 g/kg modified decoction of rehmanniae was administrated for experiment group daily, continuously for 28 days.④All rats were sacrificed at the 5^th week, BACE1 mRNA expression in hippocampus tissue was assayed by real-time quantitative reverse transcription-PCR (expressed as 2^-△△ct, Ct as cycle threshold, △Ct=Ctbace1CtGAPDH,△△Ct=△Ctinvervention groups-△Ctblank controls group).⑤Difference of measurement data among groups were compared with single factor variance analysis, and comparison between two groups was performed with LSD-t test. RESULTS: All 100 rats were adopted and every 5 rats of each group was selected to extract total RNA of hippocampal tissue, thus these 25 rats were involved in the result analysis. The BACE1 mRNA expression was significantly up-regulated in model group compared with the sham group [2^-△△ct value: (4.67±0.52), (1.07±0.08), P〈 0.01]. The BACE1 mRNA expressions in modified decoction of rehmanniae group and Donepezil group were down-regulated compared with the model group [(1.80±0.23), (1.26±0.20)]. CONCLUSION: β-amyloid1-40 injection into the bilateral hippocampus c