高胆固醇血症在阿尔茨海默病发生中的作用及柳茶提取物的治疗效应
|
摘要
目的：观察高胆固醇血症在阿尔茨海默病(AD)发生中的作用及藏药柳茶提取物的治疗效应。
方法：依数字表法将SD大鼠随机分为4组：Ⅰ为正常对照组、Ⅱ为AD模型组、Ⅲ为高胆固醇并AD组、Ⅳ为柳茶治疗组。观察各组大鼠精神、皮毛、饮食及活动等一般状况；生化法检测血和脑总胆固醇含量；Bielschowsky染色观察海马神经原纤维缠结,HE染色观察大脑皮层及海马淀粉样蛋白沉积及神经元损伤状况；实时荧光定量法检测海马及皮层组织中胆固醇代谢相关基因APOE、CYP46的表达；Western-blot法检测Tau蛋白在ser396位点磷酸化水平。
结果：与Ⅰ组相比,Ⅱ、Ⅲ组动物精神萎靡,食欲欠佳,皮毛粗糙,尤以Ⅲ组为甚,Ⅳ组精神好转,食欲增加,皮毛较顺滑。与Ⅰ组比较,Ⅱ组血胆固醇含量无显著差异,Ⅲ组明显高于Ⅱ组(P<0.05)；Ⅳ组含量均低于Ⅱ、Ⅲ组。与Ⅰ组相比Ⅱ、Ⅲ组脑胆固醇含量显著升高,Ⅲ组更为明显；柳茶治疗后Ⅳ组脑胆固醇水平下降,与Ⅲ组相比有显著性差异(P<0.05)。Ⅱ、Ⅲ组大鼠海马及皮层组织可见核深染、固缩,神经元丢失,并出现神经纤维缠结及淀粉样蛋白沉积等典型AD病理形态学变化,其中Ⅲ组大鼠变化更严重。与Ⅱ、Ⅲ组相比,Ⅳ组海马及皮层组织神经元排列相对整齐、紧密,极少见到神经纤维缠结和淀粉样蛋白沉积。与Ⅰ组相比,Ⅱ、Ⅲ组APOE mRNA表达量均增加(P<0.01),且Ⅲ组APOE mRNA表达量高于Ⅱ组(P<0.01),Ⅳ组表达量明显下降至接近Ⅰ组水平(P<0.01)；与Ⅰ组相比,Ⅱ、Ⅲ组CYP46mRNA表达量均减少(P<0.01),而Ⅳ组则明显呈高表达(P<0.01)。与Ⅰ组相比Ⅱ、Ⅲ组磷酸化的Tau蛋白表达量升高,且Ⅲ组更为明显,Ⅳ组磷酸化的Tau蛋白表达量显著下降。
结论：高胆固醇血症不仅能够促进淀粉样蛋白沉积形成老年斑,还能通过调节脑内胆固醇代谢关键基因APOE和CYP46的表达加剧Tau蛋白磷酸化形成神经纤维缠结,加重神经元丢失等AD病理学变化。柳茶提取物因有效降低了动物血和脑中胆固醇含量,故使AD大鼠脑组织中神经元丢失、淀粉样蛋白沉积以及神经纤维缠结等病理学变化得以改善,其作用机制可能与调节APOE和CYP46mRNA的表达有关。
Objective:To explore the role of Hypercholesterolemia in the outbreak of Alzheimer's disease (AD) and the treatment effect of liucha extract..
Methods:Divide the SD rats into four groups according to Figure Table, namely, normal rates for group Ⅰ, rats with Alzheimer's disease for group Ⅱ, rates with hypercholesterolemia and Alzheimer's disease for group Ⅲ, and rates treated with liucha for group Ⅳ. Then the general conditions, including spirit conditions, furs, diets and activities of the rats are observed. Biochemical methods are adopted to detect the blood and brain total cholesterol content; Bielschowsky tainting are used for observation of hippocampal neurons fiber tangles, while HE tainting are for observation of the cerebral cortex and the hippocampus dyeing amyloid deposition and neurons damage condition; Real-time quantitative fluorescence method is to detect the hippocampus and cortex in organization of cholesterol metabolism, CYP46and APOE gene expression; Western-blot method is used to detect Tau protein in ser396site phosphorylation level.
Results:Compared with group Ⅰ, group Ⅱ and Ⅲ show signs of sluggishness, poor appetite, fur roughness, which is more severe for group Ⅲ. However, Group Ⅳ is better in spirit and appetite with smooth furs. Compared with normal blood cholesterol content, group Ⅱ had no significant difference, and group Ⅲ was significantly higher than the group Ⅱ (P<0.05); group Ⅳ content are lower than group Ⅱ, Ⅲ. Compared with group Ⅰ, brain cholesterol of group Ⅱ and Ⅲ is significantly increased, with more obvious increasing in group Ⅲ; for rates treated with Liucha, brain cholesterol levels of group Ⅳ dropped, showing significant difference (P<0.05) from group Ⅲ. Rats of group Ⅱ and Ⅲshows signs of hippocampal and cortex tissue visible hyperchromatic nuclei, solid shrinkage, loss of neurons, and the emergence of nerve fibers and tangles amyloid deposition pathological morphology typical AD change, especially for group Ⅲ. Compared with group Ⅱ and Ⅲ, cortical neurons in the hippocampus organization in group Ⅳ are neat, relatively close with rare nerve fiber tangles and amyloid deposition. Compared with group Ⅰ, group Ⅱ, Ⅲ APOE mRNA express quantity increased (P<0.01), and group Ⅲ APOE mRNA express quantity is higher than group Ⅱ (P<0.01), group IV express evidently decreased to the level nearly equivalent with group Ⅰ (P<0.01); Compared with the normal group, both group Ⅱ and Ⅲ have experienced drop of CYP46mRNA express (P<0.01), while group Ⅳ see obviously high expression (P<0.01). Compared with group Ⅰ, the phosphorylated Tau protein expression volume increased for group Ⅱ and Ⅲ, while the increase in group Ⅲ is more apparent. In group Ⅳ, the phosphorylated Tau protein expression has significantly dropped.
Conclusion:Hypercholesterolemia can not only promote the amyloid deposition senile plaques form, but also adjust the brain cholesterol metabolism key genes and the expression of the CYP46and APOE, increase Tau protein formed phosphorylation entangled nerve fibers, and worsen neurons lost etc. In conclusion, it speeds up the pathological changes of the AD. The extracts of liucha reduce the cholesterol content in the blood and brain of the animals, thus improve the pathological changes for AD rats, including loss of neurons in the brain tissues, amyloid deposition and nerve fiber tangles. The mechanism has something to do with the content adjustment of the mRNA expression of CYP46and APOE.
方法：依数字表法将SD大鼠随机分为4组：Ⅰ为正常对照组、Ⅱ为AD模型组、Ⅲ为高胆固醇并AD组、Ⅳ为柳茶治疗组。观察各组大鼠精神、皮毛、饮食及活动等一般状况；生化法检测血和脑总胆固醇含量；Bielschowsky染色观察海马神经原纤维缠结,HE染色观察大脑皮层及海马淀粉样蛋白沉积及神经元损伤状况；实时荧光定量法检测海马及皮层组织中胆固醇代谢相关基因APOE、CYP46的表达；Western-blot法检测Tau蛋白在ser396位点磷酸化水平。
结果：与Ⅰ组相比,Ⅱ、Ⅲ组动物精神萎靡,食欲欠佳,皮毛粗糙,尤以Ⅲ组为甚,Ⅳ组精神好转,食欲增加,皮毛较顺滑。与Ⅰ组比较,Ⅱ组血胆固醇含量无显著差异,Ⅲ组明显高于Ⅱ组(P<0.05)；Ⅳ组含量均低于Ⅱ、Ⅲ组。与Ⅰ组相比Ⅱ、Ⅲ组脑胆固醇含量显著升高,Ⅲ组更为明显；柳茶治疗后Ⅳ组脑胆固醇水平下降,与Ⅲ组相比有显著性差异(P<0.05)。Ⅱ、Ⅲ组大鼠海马及皮层组织可见核深染、固缩,神经元丢失,并出现神经纤维缠结及淀粉样蛋白沉积等典型AD病理形态学变化,其中Ⅲ组大鼠变化更严重。与Ⅱ、Ⅲ组相比,Ⅳ组海马及皮层组织神经元排列相对整齐、紧密,极少见到神经纤维缠结和淀粉样蛋白沉积。与Ⅰ组相比,Ⅱ、Ⅲ组APOE mRNA表达量均增加(P<0.01),且Ⅲ组APOE mRNA表达量高于Ⅱ组(P<0.01),Ⅳ组表达量明显下降至接近Ⅰ组水平(P<0.01)；与Ⅰ组相比,Ⅱ、Ⅲ组CYP46mRNA表达量均减少(P<0.01),而Ⅳ组则明显呈高表达(P<0.01)。与Ⅰ组相比Ⅱ、Ⅲ组磷酸化的Tau蛋白表达量升高,且Ⅲ组更为明显,Ⅳ组磷酸化的Tau蛋白表达量显著下降。
结论：高胆固醇血症不仅能够促进淀粉样蛋白沉积形成老年斑,还能通过调节脑内胆固醇代谢关键基因APOE和CYP46的表达加剧Tau蛋白磷酸化形成神经纤维缠结,加重神经元丢失等AD病理学变化。柳茶提取物因有效降低了动物血和脑中胆固醇含量,故使AD大鼠脑组织中神经元丢失、淀粉样蛋白沉积以及神经纤维缠结等病理学变化得以改善,其作用机制可能与调节APOE和CYP46mRNA的表达有关。
Objective:To explore the role of Hypercholesterolemia in the outbreak of Alzheimer's disease (AD) and the treatment effect of liucha extract..
Methods:Divide the SD rats into four groups according to Figure Table, namely, normal rates for group Ⅰ, rats with Alzheimer's disease for group Ⅱ, rates with hypercholesterolemia and Alzheimer's disease for group Ⅲ, and rates treated with liucha for group Ⅳ. Then the general conditions, including spirit conditions, furs, diets and activities of the rats are observed. Biochemical methods are adopted to detect the blood and brain total cholesterol content; Bielschowsky tainting are used for observation of hippocampal neurons fiber tangles, while HE tainting are for observation of the cerebral cortex and the hippocampus dyeing amyloid deposition and neurons damage condition; Real-time quantitative fluorescence method is to detect the hippocampus and cortex in organization of cholesterol metabolism, CYP46and APOE gene expression; Western-blot method is used to detect Tau protein in ser396site phosphorylation level.
Results:Compared with group Ⅰ, group Ⅱ and Ⅲ show signs of sluggishness, poor appetite, fur roughness, which is more severe for group Ⅲ. However, Group Ⅳ is better in spirit and appetite with smooth furs. Compared with normal blood cholesterol content, group Ⅱ had no significant difference, and group Ⅲ was significantly higher than the group Ⅱ (P<0.05); group Ⅳ content are lower than group Ⅱ, Ⅲ. Compared with group Ⅰ, brain cholesterol of group Ⅱ and Ⅲ is significantly increased, with more obvious increasing in group Ⅲ; for rates treated with Liucha, brain cholesterol levels of group Ⅳ dropped, showing significant difference (P<0.05) from group Ⅲ. Rats of group Ⅱ and Ⅲshows signs of hippocampal and cortex tissue visible hyperchromatic nuclei, solid shrinkage, loss of neurons, and the emergence of nerve fibers and tangles amyloid deposition pathological morphology typical AD change, especially for group Ⅲ. Compared with group Ⅱ and Ⅲ, cortical neurons in the hippocampus organization in group Ⅳ are neat, relatively close with rare nerve fiber tangles and amyloid deposition. Compared with group Ⅰ, group Ⅱ, Ⅲ APOE mRNA express quantity increased (P<0.01), and group Ⅲ APOE mRNA express quantity is higher than group Ⅱ (P<0.01), group IV express evidently decreased to the level nearly equivalent with group Ⅰ (P<0.01); Compared with the normal group, both group Ⅱ and Ⅲ have experienced drop of CYP46mRNA express (P<0.01), while group Ⅳ see obviously high expression (P<0.01). Compared with group Ⅰ, the phosphorylated Tau protein expression volume increased for group Ⅱ and Ⅲ, while the increase in group Ⅲ is more apparent. In group Ⅳ, the phosphorylated Tau protein expression has significantly dropped.
Conclusion:Hypercholesterolemia can not only promote the amyloid deposition senile plaques form, but also adjust the brain cholesterol metabolism key genes and the expression of the CYP46and APOE, increase Tau protein formed phosphorylation entangled nerve fibers, and worsen neurons lost etc. In conclusion, it speeds up the pathological changes of the AD. The extracts of liucha reduce the cholesterol content in the blood and brain of the animals, thus improve the pathological changes for AD rats, including loss of neurons in the brain tissues, amyloid deposition and nerve fiber tangles. The mechanism has something to do with the content adjustment of the mRNA expression of CYP46and APOE.
引文
[1]方传勤,周华东,李敬诚等.高胆固醇饮食对AD模型大鼠空间学习和记忆的影响[J].重庆医学.2007,36(9)：849-851.
[2]秀梅.正常增龄与AD患者额叶脑组织的比较蛋白质组学[博士论文].北京：解放军总医院军医进修学院.2009.
[3]姚秀卿,刘蓉,王建枝.阿尔茨海默病的临床诊断研究进展[J].神经损伤与功能重建.2007,2(4)：233-236.
[4]Alessandro Peri,Mario Serio. Neuroprotective effects of the Alzheimer's Disease-related gene seladin-1[J]. Journal of Molecular Endocrinology.2008,41:251-261
[5]Fabricio A, Pamplona, Pablo Pandolfo, Filipe S. Duarte, Altered emotionality leads to increased pain tolerance in amyloid β(Aβ1-40)peptide-treated mice[J].Behavioural Brain Research.2010,96-102.
[6]刘英,高璐,马丽香.APP/PS1双转基因小鼠大脑皮质老年斑的病变对其学习记忆能力的影响[J].神经解剖学杂志.2007,23(3)：225-231.
[7]朱莹莹.拟老年痴呆大鼠模型嗅觉系统相关改变的研究[博士生论文].北京：北京协和医学院.2008.
[8]方传勤.高胆固醇对AD大鼠认知功能影响及机制[博士论文].重庆：第三军医大学.2007.
[9]Ramirez DM,Andersson S,Russell DW. Neuronal expression and subcellular localization of cholesterol 24-hydroxylase in the mouse brain [J]. The Journal Of Comparative Neurology.2008,507(5):1676-93.
[10]姚妍怡,刘志国,屈伸.载脂蛋白E3及其突变体apoE2、apoE4的克隆及表达[J].华中科技大学学报.2004,33(2)：109-112.
[11]刘倩,吴为辉,李人望,等.载脂蛋白E4与阿尔滋海默病的相关研究进展[J].化学进展.2007,19(12)：2007-2011.
[12]许晶.胆固醇对早老性痴呆转基因动物模型APP加工及Aβ生成的影响[硕十论文].北京：中国协和医科大学.2006.
[13]Kabbara A,Payet N,Cottel D,etal.Exclusion of CYP46 and APOM as candidate genes for Alzheimer's disease in a French population[J].Neurosci Lett.2004,363(2):139-43.
[14]Ramirez DM,Andersson S,Russell DW. Neuronal expression and subcellular localization of cholesterol 24-hydroxylase in the mouse brain [J]. The Journal Of Comparative Neurology.2008,507(5):1676-93.
[15]Yoshihiko Ohyama, Steve Meaney, Maura Heverin,etal. Studies on the Transcriptional Regulation of Cholesterol 24-Hydroxylase (CYP46A1) [J]. The Journal Of Biological Chemistry.2006,281 (7):3810-3820.
[16]米东华,胆固醇代谢与阿尔茨海默病、帕金森痴呆的关系研究[硕十论文].天津：天津医科大学.2007.
[17]陈付艳,聂坤,于建春,韩景献.胆固醇代谢异常与阿尔兹海默氏症[J].现代生物医学进展.2007,7(11)：1758-1760.
[18]黎凯.锋阿尔茨海默病与CYP19基因、谷胱甘肽S-转移酶基因多态性的相关性分析[硕士论文].广州.南方医科大学.2009.
[19]陶婷婷,柏川,王天志,等.藏药柳茶挥发油化学成分的研究陶[J].华西药学杂志.2004,19(3)：178-179.
[20]武燕,武三卯.藏药柳茶提取物对异烟肼利福平合用引起小鼠肝损伤的保护作用[J].中国医院药学杂志.2010,30(19)：1660-1663.
[21]张承忠,封士兰,李冲,等.藏药柳茶中的三萜类化学成分[J].中国中药杂志.1992,6：360-373.
[22]张承忠,李冲,封士兰,等.鲜卑花酯的分离和结构[J].药学学报.1993,10：798-800.
[23]游元元,柏川,王天志.藏药柳茶的化学成分研究[J].华西药学杂志.2004,19(2)：113-114.
[24]陈璐,陶婷婷,王天志RP-HPLC测定藏药柳茶中阿魏酸和异阿魏酸的含量[J].华西药学杂志.2006,21(2)：196-197.
[25]石磊,张冬梅,等.柳茶多糖的分离纯化及气相色谱-质谱分析[J].时珍国医国药.2009,20(9)：2191-2192.
[26]刘昕,等.柳茶提取物对LIBP小鼠免疫功能影响的实验观察[J].兰州医学院学报.1994,20(2)：146.
[27]刘昕,潘兴斌.等.柳茶对小鼠抗氧化能力及免疫器官影响的实验观察[J].兰州医学院学报.1994；20(3)：143-145.
[28]潘兴斌,刘昕,等.柳茶提取物对小鼠巨噬细胞吞噬功能及对白细胞-2介素产生的影响[J].兰州医学院学报.1995,21(4)：201-202.
[29]刘昕,潘兴斌等.柳茶提取物对正常小鼠血SOD、GSH-Px及MDA含量的影响[J].中医院学报.1995,6：35-36.
[30]刘昕,杨萍,潘兴斌.柳茶提取物免疫调节作用的实验研究[J].免疫学杂志.1996,12(4)：237-240.
[31]刘听,潘兴斌,李兴玉.藏药柳茶提取物对K562肿瘤细胞生长的影响[J].中国病理生理杂志.2003；19(2)：253-255.
[32]雒或,刘昕.柳茶对荷瘤小鼠脾淋巴细胞产生IL-2和TNF-α的影响[J].中药药理与临床.2007,23(2)：42-43.
[33]马晓燕,王晶宇,卢岩松,等.藏药柳茶提取物对非酒精性脂肪肝病大鼠肝脏脂肪代谢的影响[J].世界华人消化杂志.2010.18(26)：2739-2744.
[34]汪晓立,郭继明,杨兵勋.柳茶调节脂类代谢作用的研究.[J]中草药.1995,(09)：473-475+504.
[35]汪晓立,杨兵勋.柳茶对CCl-4月肝损伤的保护作用及机理初探.[J]中国中药杂志.1995,(12)：746-748.
[36]姚莉,焦海胜,李晓强.柳茶对高脂血症大鼠血脂、超氧化物歧化酶及丙二醛的影响.[J]兰州大学学报.2010,36(2)：68-70.
[37]姚莉,骆勤,李红卫,等.柳茶对D-半乳糖胺肝损伤的保护作用及机制.[J]中国医院药学杂志.2010,30(6)：483-484
[38]Sun SW, Yu HQ, Zhang H, et al,Quercetin attenuates spontaneous behavior and spatial memory impairment in D-galactose-treated mice by increasing brain antioxidant capacity[J]. Nutr Res,2007; 27:169-75.
[39]吴敏霞,谢捷明,俞昌喜.Tau蛋白异常磷酸化与阿尔茨海默病的关系[J].海峡药学.2004,16(2)：17-20.
[40]陈莉,刘小艳,黄日越.鸟圆补血口服液对D—半乳糖和亚硝酸钠所致痴呆模型小鼠生化指标的影响[J].中国老年学杂志.2009,5(29)：1206-1208.
[41]罗焕敏.陈子晟.一种新的老年痴呆动物模型[J].中国老年学杂志.2003,3(23)：179-182.
[42]陈玉静,田金洲.中药复方金思维对拟AD模型大鼠海马神经元内微管结构和tau蛋白磷酸 化相关酶类.表达的影响[J].天津中医药,2008,25(1)：59-62.
[43]孙秀兰,谢桂琴.红景天素改善D-半乳糖-Aβ脑内注射老年痴呆模型鼠的行为障碍[J].基础医学与临床.2003,3(6)：620-624.
[44]Lesser GT,Beeri MS,Schmeidler J, et al.Cholesterol and LDL Relate to Neuritic Plaques and to APOE4 Presence but Not to Neurofibrillary Tangles[J]. Current Alzhelimer Research.2011,8(3):303-312.
[45]李晨虎,李春波,吴文源.脑血管病变与阿尔茨海默病关系的研究进展[J].中华神经科杂志.2004,5(37)：121-123.
[46]Kalaria RN.Small vessel disease and Alzheimer's dementia Pathological considerations[J].Cerebrovase Dis.2002,13(2):48-52.
[47]姜希娟,郭茂娟,苏金玲,等.银杏内酯B对胆固醇和载脂蛋白E4损伤海马神经元胆固醇代谢的影响[J].中国老年学杂志.2009,29：2638-2640.
[48]张瑞峰马,东明姜希,娟卢斌,苏金等.辛伐他汀对铜和高胆固醇阿尔茨海默病家兔海马淀粉样肽前蛋白mRNA表达的影响[J].中国老年学杂志.2011,31(3)：443-447
[49]朱海暴,石如玲,赵朝贤.血胆固醇与脑胆固醇关系的研究[J].中国老年学杂志.2005,10(25)：1208-1210.
[50]JiekH,ZombergGL,Jiek55,etal.Statinsandtheriskofdementia[J].Lancet.2000,356:162-1631.
[2]秀梅.正常增龄与AD患者额叶脑组织的比较蛋白质组学[博士论文].北京：解放军总医院军医进修学院.2009.
[3]姚秀卿,刘蓉,王建枝.阿尔茨海默病的临床诊断研究进展[J].神经损伤与功能重建.2007,2(4)：233-236.
[4]Alessandro Peri,Mario Serio. Neuroprotective effects of the Alzheimer's Disease-related gene seladin-1[J]. Journal of Molecular Endocrinology.2008,41:251-261
[5]Fabricio A, Pamplona, Pablo Pandolfo, Filipe S. Duarte, Altered emotionality leads to increased pain tolerance in amyloid β(Aβ1-40)peptide-treated mice[J].Behavioural Brain Research.2010,96-102.
[6]刘英,高璐,马丽香.APP/PS1双转基因小鼠大脑皮质老年斑的病变对其学习记忆能力的影响[J].神经解剖学杂志.2007,23(3)：225-231.
[7]朱莹莹.拟老年痴呆大鼠模型嗅觉系统相关改变的研究[博士生论文].北京：北京协和医学院.2008.
[8]方传勤.高胆固醇对AD大鼠认知功能影响及机制[博士论文].重庆：第三军医大学.2007.
[9]Ramirez DM,Andersson S,Russell DW. Neuronal expression and subcellular localization of cholesterol 24-hydroxylase in the mouse brain [J]. The Journal Of Comparative Neurology.2008,507(5):1676-93.
[10]姚妍怡,刘志国,屈伸.载脂蛋白E3及其突变体apoE2、apoE4的克隆及表达[J].华中科技大学学报.2004,33(2)：109-112.
[11]刘倩,吴为辉,李人望,等.载脂蛋白E4与阿尔滋海默病的相关研究进展[J].化学进展.2007,19(12)：2007-2011.
[12]许晶.胆固醇对早老性痴呆转基因动物模型APP加工及Aβ生成的影响[硕十论文].北京：中国协和医科大学.2006.
[13]Kabbara A,Payet N,Cottel D,etal.Exclusion of CYP46 and APOM as candidate genes for Alzheimer's disease in a French population[J].Neurosci Lett.2004,363(2):139-43.
[14]Ramirez DM,Andersson S,Russell DW. Neuronal expression and subcellular localization of cholesterol 24-hydroxylase in the mouse brain [J]. The Journal Of Comparative Neurology.2008,507(5):1676-93.
[15]Yoshihiko Ohyama, Steve Meaney, Maura Heverin,etal. Studies on the Transcriptional Regulation of Cholesterol 24-Hydroxylase (CYP46A1) [J]. The Journal Of Biological Chemistry.2006,281 (7):3810-3820.
[16]米东华,胆固醇代谢与阿尔茨海默病、帕金森痴呆的关系研究[硕十论文].天津：天津医科大学.2007.
[17]陈付艳,聂坤,于建春,韩景献.胆固醇代谢异常与阿尔兹海默氏症[J].现代生物医学进展.2007,7(11)：1758-1760.
[18]黎凯.锋阿尔茨海默病与CYP19基因、谷胱甘肽S-转移酶基因多态性的相关性分析[硕士论文].广州.南方医科大学.2009.
[19]陶婷婷,柏川,王天志,等.藏药柳茶挥发油化学成分的研究陶[J].华西药学杂志.2004,19(3)：178-179.
[20]武燕,武三卯.藏药柳茶提取物对异烟肼利福平合用引起小鼠肝损伤的保护作用[J].中国医院药学杂志.2010,30(19)：1660-1663.
[21]张承忠,封士兰,李冲,等.藏药柳茶中的三萜类化学成分[J].中国中药杂志.1992,6：360-373.
[22]张承忠,李冲,封士兰,等.鲜卑花酯的分离和结构[J].药学学报.1993,10：798-800.
[23]游元元,柏川,王天志.藏药柳茶的化学成分研究[J].华西药学杂志.2004,19(2)：113-114.
[24]陈璐,陶婷婷,王天志RP-HPLC测定藏药柳茶中阿魏酸和异阿魏酸的含量[J].华西药学杂志.2006,21(2)：196-197.
[25]石磊,张冬梅,等.柳茶多糖的分离纯化及气相色谱-质谱分析[J].时珍国医国药.2009,20(9)：2191-2192.
[26]刘昕,等.柳茶提取物对LIBP小鼠免疫功能影响的实验观察[J].兰州医学院学报.1994,20(2)：146.
[27]刘昕,潘兴斌.等.柳茶对小鼠抗氧化能力及免疫器官影响的实验观察[J].兰州医学院学报.1994；20(3)：143-145.
[28]潘兴斌,刘昕,等.柳茶提取物对小鼠巨噬细胞吞噬功能及对白细胞-2介素产生的影响[J].兰州医学院学报.1995,21(4)：201-202.
[29]刘昕,潘兴斌等.柳茶提取物对正常小鼠血SOD、GSH-Px及MDA含量的影响[J].中医院学报.1995,6：35-36.
[30]刘昕,杨萍,潘兴斌.柳茶提取物免疫调节作用的实验研究[J].免疫学杂志.1996,12(4)：237-240.
[31]刘听,潘兴斌,李兴玉.藏药柳茶提取物对K562肿瘤细胞生长的影响[J].中国病理生理杂志.2003；19(2)：253-255.
[32]雒或,刘昕.柳茶对荷瘤小鼠脾淋巴细胞产生IL-2和TNF-α的影响[J].中药药理与临床.2007,23(2)：42-43.
[33]马晓燕,王晶宇,卢岩松,等.藏药柳茶提取物对非酒精性脂肪肝病大鼠肝脏脂肪代谢的影响[J].世界华人消化杂志.2010.18(26)：2739-2744.
[34]汪晓立,郭继明,杨兵勋.柳茶调节脂类代谢作用的研究.[J]中草药.1995,(09)：473-475+504.
[35]汪晓立,杨兵勋.柳茶对CCl-4月肝损伤的保护作用及机理初探.[J]中国中药杂志.1995,(12)：746-748.
[36]姚莉,焦海胜,李晓强.柳茶对高脂血症大鼠血脂、超氧化物歧化酶及丙二醛的影响.[J]兰州大学学报.2010,36(2)：68-70.
[37]姚莉,骆勤,李红卫,等.柳茶对D-半乳糖胺肝损伤的保护作用及机制.[J]中国医院药学杂志.2010,30(6)：483-484
[38]Sun SW, Yu HQ, Zhang H, et al,Quercetin attenuates spontaneous behavior and spatial memory impairment in D-galactose-treated mice by increasing brain antioxidant capacity[J]. Nutr Res,2007; 27:169-75.
[39]吴敏霞,谢捷明,俞昌喜.Tau蛋白异常磷酸化与阿尔茨海默病的关系[J].海峡药学.2004,16(2)：17-20.
[40]陈莉,刘小艳,黄日越.鸟圆补血口服液对D—半乳糖和亚硝酸钠所致痴呆模型小鼠生化指标的影响[J].中国老年学杂志.2009,5(29)：1206-1208.
[41]罗焕敏.陈子晟.一种新的老年痴呆动物模型[J].中国老年学杂志.2003,3(23)：179-182.
[42]陈玉静,田金洲.中药复方金思维对拟AD模型大鼠海马神经元内微管结构和tau蛋白磷酸 化相关酶类.表达的影响[J].天津中医药,2008,25(1)：59-62.
[43]孙秀兰,谢桂琴.红景天素改善D-半乳糖-Aβ脑内注射老年痴呆模型鼠的行为障碍[J].基础医学与临床.2003,3(6)：620-624.
[44]Lesser GT,Beeri MS,Schmeidler J, et al.Cholesterol and LDL Relate to Neuritic Plaques and to APOE4 Presence but Not to Neurofibrillary Tangles[J]. Current Alzhelimer Research.2011,8(3):303-312.
[45]李晨虎,李春波,吴文源.脑血管病变与阿尔茨海默病关系的研究进展[J].中华神经科杂志.2004,5(37)：121-123.
[46]Kalaria RN.Small vessel disease and Alzheimer's dementia Pathological considerations[J].Cerebrovase Dis.2002,13(2):48-52.
[47]姜希娟,郭茂娟,苏金玲,等.银杏内酯B对胆固醇和载脂蛋白E4损伤海马神经元胆固醇代谢的影响[J].中国老年学杂志.2009,29：2638-2640.
[48]张瑞峰马,东明姜希,娟卢斌,苏金等.辛伐他汀对铜和高胆固醇阿尔茨海默病家兔海马淀粉样肽前蛋白mRNA表达的影响[J].中国老年学杂志.2011,31(3)：443-447
[49]朱海暴,石如玲,赵朝贤.血胆固醇与脑胆固醇关系的研究[J].中国老年学杂志.2005,10(25)：1208-1210.
[50]JiekH,ZombergGL,Jiek55,etal.Statinsandtheriskofdementia[J].Lancet.2000,356:162-1631.