摘要
目的：以能量代谢障碍淀粉样前体蛋白（APP）/早老素基因1（PS1）转基因小鼠为阿尔茨海默病（Alzheimer＇s disease,AD）模型,探讨地黄饮子通过调控蛋白激酶R样内质网激酶（protein kinase r-like endoplasmic reticulum kinase,PERK）,真核细胞起始因子2α（eukaryotic initiation factor 2α,eI F2α）通路抑制内质网应激导致的β-淀粉样蛋白（Aβ）累积的作用机制。方法：4月龄APP/PS1转基因小鼠随机分为正常组、模型组、阳性药（安理申）组、地黄饮子低、中、高剂量组。除正常组小鼠腹腔注射无菌生理盐水外,其余各组小鼠均腹腔注射100 mg·kg-13-硝基丙酸（3-nitropropionic acid,3-NP） 1次。小鼠造模后,立即给药。灌胃给药1周后,Morris水迷宫实验检测小鼠学习记忆能力,高效液相色谱法检测小鼠脑内三磷酸腺苷（ATP）,二磷酸腺苷（ADP）,一磷酸腺苷（AMP）含量,并计算脑能荷（energy charge,EC）。蛋白免疫印迹法（Western blot）检测小鼠脑内PERK,eI F2α磷酸化水平,β-淀粉样前体蛋白水解酶1（BACE1）表达水平。实时荧光定量聚合酶链式反应（Real-time PCR）检测BACE1 mRNA表达水平。酶联免疫吸附测定法（ELISA）检测小鼠脑内Aβ含量。结果：与正常组比较,Morris水迷宫实验结果显示,在定位巡航实验中,地黄饮子可以明显缩短模型小鼠的逃避潜伏期（P 〈0. 05,P 〈0. 01）;空间探索实验中,地黄饮子可以增加模型小鼠穿越目标区域的次数（P 〈0. 05,P 〈0. 01）和在目标象限的停留时间（P 〈0. 01）,减少相对象限的停留时间（P 〈0. 05,P 〈0. 01）。地黄饮子可以显著降低AMP水平,明显升高ATP,ADP水平,显著提高模型小鼠脑内EC水平（P 〈0. 05,P 〈0. 01）,抑制PERK,eI F2α的磷酸化（P 〈0. 01）和BACE1的蛋白水平（P 〈0. 01）,但对BACE1 mRNA转录没有显著影响。地黄饮子能够减少模型小鼠脑内Aβ的含量（P 〈0. 01）。结论：�
Objective： To explore the effect and mechanism of Dihuang Yinzi on β-amyloid protein（ Aβ）accumulation via protein kinase r-like endoplasmic reticulum kinase（ PERK）,eukaryotic initiation factor 2α（ e IF2α） pathway in energy inhibited APP/PS1 transgenic Alzheimer＇s disease（ AD） mice. Method： APP/PS1 transgenic mice at 4-month-old were randomly divided into normal control group,model group,positive drug group（ Aricept）,Dihuang Yinzi low-,medium-and high-dose groups. Mice were intraperitoneally injected with100 mg·kg-13-nitropropionic acid（ 3-NP） except those in normal control group（ intraperitoneally injected with normal saline） to establish models. Drug administration was given immediately modeling,and after 1 week of intragastric administration,the learning and memory ability of mice was tested by Morris water maze test; the contents of adenosine triphosphate（ ATP）,adenosine diphosphate（ ADP）,and adenosine monophosphate（ AMP）in the brain of mice were detected by high performance liquid chromatography（ HPLC）; and the energy charge（ EC） was calculated. Western blot was performed to detect the levels of PERK,e IF2α phosphorylation andβ-amyloid precursor protein hydrolase 1（ BACE1） expression in the brain of mice. The transcriptional level of BACE1 gene was detected by Real-time fluorescence quantitative polymerase chain reaction（ PCR）. The content of cerebral Aβ was detected by enzyme-linked immunosorbent assay（ ELISA）. Result： Morris water maze test showed that as compared with the normal group,Dihuang Yinzi can significantly shorten the escape latency of model mice in navigation test（ P 〈 0. 05, P 〈 0. 01）. In spatial probe test,Dihuang Yinzi can increase the number passing the target area（ P 〈 0. 05,P 〈 0. 01） and the residence time in target quadrant（ P 〈 0. 01）,reduce the residence time in opposite quadrant（ P 〈 0. 05,P 〈 0. 01）. Dihuang Yinzi can significantly decrease AMP level,increase ATP and ADP levels,
作者
张志辰
温彬宇
高俊峰
唐旭
黄倩倩
马涛
ZHANG Zhi-chen;WEN Bin-yu;GAO Jun-feng;TANG Xu;HUANG Qian-qian;MA Tao(Dongfang Hospital,Beifing University of Chinese Medicine,Beijing 100078,China;The Third Affiliated Hospital,Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2018年第21期91-98,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81673929)
