【作者】 牛英才；

【导师】 谢宁；

【作者基本信息】 黑龙江中医药大学 ， 中医基础理论， 2005， 博士

【摘要】 目的 探讨地黄饮子对D-gal致亚急性衰老合并海马注射Aβ₁-40建立的AD大鼠模型作用的分子机制。 方法:18月龄Wistar雄性大鼠120只随机分为地黄饮子预防组、地黄饮子治疗组、模型组、盐酸多奈哌齐组、假手术组和老年对照组,另以16只4月龄Wistar大鼠作为青年对照组。地黄饮子预防组、地黄饮子治疗组、模型组、盐酸多奈哌齐组先腹腔注射D-gal 28天,同时预防组给予地黄饮子灌胃给药,然后双侧海马注射Aβ₁-40,制备复合AD模型。注射后3天起,地黄饮子治疗组、盐酸多奈哌齐对照组给药28天后检测各项指标。通过Morris水迷宫试验测试大鼠的空间学习记忆能力的变化,光镜、电镜观察海马组织形态结构变化,化学比色法测定CAT、GSH-Px、MAO活性变化,免疫组织化学染色法检测NMDAR、NOS、ChAT的表达改变,核酸原位杂交法检测海马组织Tau蛋白mRNA表达变化,RT-PCR法检测APP mRNA、SOD1 mRNA和ICE mRNA的表达变化,以及地黄饮子对上述指标的影响。 结果:行为学结果显示模型组与老年对照组比较,潜伏期延长,穿过平台次数明显减少,有显著性差异;地黄饮子预防组、治疗组与模型组比较,潜伏期缩短,穿过平台次数明显增多,有显著性差异。电镜、光镜观察结果显示模型组大鼠海马神经元减少和变性;地黄饮子能改善AD大鼠海马病理改变。模型组与老年对照组比较,脑组织CAT、GSH-Px的活性明显降低,有显著性差异;地黄饮子预防组、治疗组与模型组比较,CAT、GSH-Px活性升高,有显著性差异。与老年对照组比较,模型组MAO活力升高,有显著性差异;地黄饮子预防组、治疗组与模型组比较,MAO活性降低,有显著性差异。与老年对照组比较,模型组大鼠脑组织中ChAT、NR2B、nNOS的面密度和PU值均明显降低,有显著性差异;预防组、治疗组与模型组比较,地黄饮子能提高ChAT、NR2B、nNOS的表达,有显著性差异。与老年对照组比更多还原

【Abstract】 Objective:the aim of our study was to investigate protective and therapeutic mechanism of Rehmannia Decotion in experimental AD rats.Methods:Male , Wistar 18 month Rat（n=120） were randomly divided into six groups.To induce AD,four groups rats received intraperitoneral injection of D-galactose combined with amyloid-β_1-40 injection the bilateral Hippocampus.And the left, one goup served as blank controls ,the other as sham operated group.4 month rat （n=16） served as youth bank control, Rehmannia Decotion was administered by oral gavage to protective group before AD induction and therapeutic group after AD induction. Donepezil Hydrochloride Tablets served as control drug. Every indice was studied after 4 week AD induction.Result:In the behavior experiment ,model group rats’ Escape latency was longest and number of times of pass safe platform was more than blank control group rats,white protective and therapeutic groups of Rehmannia Decotion rats were shorter and number of times pass safe platform was less than model group rats. By the light microscopy and electron microscopy observation,The neurocyte decreased and deformed in model group rats. Rehmannia Decotion can ameliorate this pathological changes. Measuring the activity of enzyme by chemical colorimetric method,we could detect that the activity of catalase、 Glutathione peroxidase was lower in AD model group rats compared with blank control group rats , and these alterations were prevented by Rehmannia Decotion treatment.The activity of monoamine oxidase was higher in AD model group rats than blank control group rats , and this alterations were prevented by Rehmannia Decotion treatment.CMIAS image system was used for cover density,and PU value analysis.Detected by immunohistochemistry,the contents of Chat, nNOS and NMDAR2B protein in AD model group rats obviously decreased compared with blank control group rats. And these alterations were prevented by Rehmannia Decotion treatment.We used acid hybridization in situ in detection the expression of Tau mRNA , Tau mRNA in the Hippocampus in AD model group rats obviously increased compared with blank control group rats. And this alteration was prevented by Rehmannia Decotion treatment. Semiquantitative reverse transcription Polymerase chain reaction amplification of APP and glyceraldehyde3-phosphate dehydrogenase transcription performed. Densitometric analysis of PCR products showed that APP mRNA was significantly higher in model group rats than blank control group, protective and therapeutic groups of Rehmannia Decotion rats.APP mRNA levels were lower than model group rats,RT-PCR also was performed for SOD and ICE mRNA expression,significant difference was found in SOD and ICE amplification products between AD rats and blank control rats,Between protective and therapeutic groups of Rehmannia Decotion rats and model group rats. ICE mRNA expression in model group increased compared with blank control group.ICE mRNA expression in protective and therapeutic groups of Rehmannia Decotion rats decreased compared with Model group. SOD1 mRNA expression in model group decreased compared with blank control group.SOD1 mRNA expression in protective and therapeutic groups of Rehmannia Deco-tion rats increase compared with Model group.Conclusion.’These results suggest that AD rat model could be induced by intraperitoneral injection of D-galactose combined with amyloid-Pi.40 injection the Hippocampus.Rehmannia Decotion possesses a certain improvement of the ChAT,nNOS and NR2B content,decreased the MAO activity,consequently increased the learning memory level,and could decrease Tau mRNA expression. Rehmannia Decotion possesses a certain improvement of CAT and GSH-Px activity.Up-regulateing the SOD1 mRNA expression.According alleviating the damage of free radical,Rehmannia Decotion could downregulate Tau,ICE and APP mRNA expression, decrease the amount of senile plaques, NFT and Apoptosis in their brains. Kidney-essence weak, phelegm is the basic etiological factor for AD,And Replenishing kidney esse-更多还原