地黄饮子影响阿尔茨海默病模型大鼠大脑单胺氧化酶B基因的表达

Effect of Dihuang Yinzi on expression of brain monoamine B gene in rat models of Alzheimer disease

目的:研究地黄饮子对阿尔茨海默病模型大鼠大脑单胺氧化酶B基因表达的影响。方法:实验于2005-11/12在黑龙江中医药大学中医基础实验室完成。实验选用普通级Wistar雄性大鼠120只,随机分为老年对照组、假手术组、模型组、盐酸多奈哌齐组、地黄饮子预防组和地黄饮子治疗组,每组20只,另选普通级,鼠龄4个月的Wistar大鼠15只,作为青年对照组。地黄饮子浓缩浸膏配成含生药0.26g/mL的混悬液,盐酸多奈哌齐片配成0.0862g/L的混悬液。以D-半乳糖致亚急性衰老合并海马注射β-淀粉样蛋白1-40建立阿尔茨海默病大鼠模型,假手术组大鼠只注射生理盐水,老年对照组和青年对照组大鼠不给予手术处理,采用逆转录聚合酶链式反应技术研究模型大鼠脑组织单胺氧化酶B信使核糖核酸表达。结果:35只大鼠纳入结果分析。总核糖核酸电泳凝胶图显示,5S、18S、28S条带完整、清晰,并且28S带的宽度及亮度约是18S的两倍,证明提取的核糖核酸可作为逆转录反应的模板。模型组大鼠脑组织单胺氧化酶B信使核糖核酸表达增多(0.4948±0.0316),与青年对照组(0.0683±0.0196)、老年对照组(0.1864±0.0256)、假手术组(0.1885±0.0266)比较差异有显著性(P均<0.01)。假手术组与老年对照组比较差异无显著性意义(P>0.05)。与模型组相比较,地黄饮子预防组(0.3678±0.0222)、地黄饮子治疗组(0.3882±0.0205)、盐酸多奈哌齐组(0.3774±0.0311)单胺氧化酶B信使核糖核酸的表达明显降低(P<0.01)。但同时地黄饮子预防组、地黄饮子治疗组与盐酸多奈哌齐组比较,单胺氧化酶B信使核糖核酸表达差异无显著性(P>0.05)。结论:地黄饮子能下调阿尔茨海默病模型大鼠大脑单胺氧化酶B信使核糖核酸的表达。

AIM： To study the effect of Dihuang Yinzi （DHYZ） on expression of brain monoamine oxidase （MAO） B mRNA in rats with Alzheimer disease （AD）. METHODS： The experiment was finished in the fundamental theory laboratory of Heilongjiang University of Traditional Chinese Medicine from November to December in 2005. Totally 120 common male Wistar rats of 4 months old were randomly divided into old control group, sham operated group, model group, Donepezil Hydrochloride Tablets group, protective group of DHYZ and therapeutic group of DHYZ, with 20 rats in every group. And 15 common male Wistar rats of 4 months old were served as youth control group. Concentrated extracts of DHYZ were dispensed into suspension containing 0.26 g/mL crude drug, and Donepezil Hydroehloride Tablets were dispensed into suspension containing 0.086 2 g/L crude drug. AD model was established with D-galactose, which caused subacute aging combined with hippocampus injection of β-amyloid 1-40, while physiological saline was injected into the rats of sham operated group, and no operation was conducted in old control group and youth control group. Expression of brain MAO-B mRNA in the model was observed by reverse transcription-polymerase chain reaction （RT-PCR）. RESULTS： Thirty-five rats were included in the results analysis. The picture of electrephoresis jelly in RNA indicated： The bands of 5 S, 18 S and 28 S were intact and clear, the width and brightness of 28 S band were double to that of 18 S, which proved the extracted RNA might be served as the stent of RT. Compared with youth control group （0.068 3±0.019 6）, old control group （0.186 4±0.025 6） and sham operated group （0.188 5±0.026 6）, the expression of MAO-B mRNA increased in model group （0.494 8±0.031 6）, with the significant difference （P 〈 0,01）. There was no significant difference between sham operated group and old control group （P 〉 0.05）. Compared with model group, the expression of brain MAO-B mRNA obviously decreased in DHYZ protective group （0：367 8±0.022 2）, DHYZ therapeutic group （0.388 2±0.020 5） and Donepezil Hydrochloride Tablets group （0.377 4±0.031 1）, （P 〈 0.01）. But no significant difference was found in protective group of DHYZ, therapeutic group of DHYZ and Donepezil Hydroehloride Tablets group （P 〉 0.05）. CONCLUSION： DHYZ can decrease the expression of brain MAO-B mRNA in AD rats.