摘要
目的研究加减薯蓣丸对APP/PS1模型小鼠的保护作用及可能机制。方法选取20只5月龄APP/PS1小鼠分为模型组、中药组,每组10只,另设同窝野生型小鼠10只为对照组,中药组给予加减薯蓣丸每日14g/kg干预4周,其余给予等容积生理盐水灌胃,Morris水迷宫检测学习记忆能力,ELISA检测海马组织β淀粉样蛋白42(amyloid-β42,Aβ42)水平,Western blot检测海马区微管相关蛋白3(microtubuleassociated light chain3,LC3)-Ⅱ/LC3-Ⅰ、P62蛋白、组织蛋白酶B(cathepsin B,CTSB)、溶酶体膜相关蛋白1(lysosomal-associated membrane proteins 1,LAMP1)、总转录因子EB(transcription factor EB,TFEB)、核内TFEB蛋白水平。结果与对照组相比,模型组小鼠逃避潜伏期延长(P<0.05),穿越平台次数及在原平台象限停留时间占比均显著减少(P<0.05);与模型组相比,中药组小鼠逃避潜伏期缩短(P<0.05),穿越平台次数及在原平台象限停留时间占比明显增多(P<0.05)。与对照组相比,模型组小鼠海马组织中Aβ42含量显著升高,LC3-Ⅱ/LC3-Ⅰ比值显著升高,P62蛋白、细胞核内TFEB和总TFEB表达水平均显著上调(P<0.05),LAMP1、CTSB蛋白表达水平均显著下调(P<0.05);与模型组比较,中药组小鼠海马组织中Aβ42含量显著降低(P<0.05),LAMP1、CTSB、细胞核内TFEB表达水平均显著上调(P<0.05),P62蛋白表达水平显著下调(P<0.05),LC3-Ⅱ/LC3-Ⅰ比值、总TFEB水平无明显变化(P>0.05)。结论加减薯蓣丸通过促进TFEB核转移增强自噬溶酶体途径,减少海马区Aβ42水平,改善APP/PS1模型小鼠学习记忆能力。
Objective To investigate the protective effect of Modified Dioscorea opposita Thunb.Pill on APP/PS1 mice and the possible mechanism.Methods A total of 20 APP/PS1 mice,aged 5 months,were divided into model group and traditional Chinese medicine(TCM)group,with 10 mice in each group,and 10 wild-type littermates were selected as control group.The mice in the TCM group were given the intervention with Modified Dioscorea opposita Thunb.Pill at a dose of 14 g/(kg·d)for 4 weeks,and those in the other two groups were given an equal volume of normal saline.Morris water maze was used to evaluate the learning and memory abilities;ELISA was used to measure the level of amyloid-β42(Aβ42)in the hippocampus;Western blot was used to measure the protein expression of microtubule-associated light chain 3(LC3)-Ⅱ/LC3-Ⅰ,sequestosome 1(P62),cathepsin B(CTSB),lysosomal-associated membrane proteins 1(LAMP1),total transcription factor EB(TFEB),and nuclear TFEB in the hippocampus.Results Compared with the control group,the model group had a significant increase in escape latency(P<0.05)and significant reductions in the number of platform crossings and the percentage of time spent in the original quadrant(P<0.05).Compared with the model group,the TCM group had a significant reduction in escape latency(P<0.05)and significant increases in the number of platform crossings and the percentage of time spent in the original quadrant(P<0.05).Compared with the control group,the model group had significant increases in Aβ42 level in the hippocampus,LC3-Ⅱ/LC3-Ⅰratio,and protein expression of P62,nuclear TFEB,and total TFEB(P<0.05),as well as significant reductions in the protein expression of LAMP1 and CTSB(P<0.05).Compared with the model group,the TCM group had a significant reduction in Aβ42 level in the hippocampus(P<0.05),significant increases in the protein expression of LAMP1,CTSB,and nuclear TFEB(P<0.05),and a significant reduction in the protein expression of P62(P<0.05),with no significant changes in LC3-Ⅱ/LC3-Ⅰratio and total TFEB(P>0.05).Conclusion Modified Dioscorea opposita Thunb.Pill can improve the learning and memory abilities of APP/PS1 mice by promoting TFEB nuclear transfer,enhancing the autophagy-lysosomal pathway,and reducing the level of Aβ42 in the hippocampus.
作者
邱静
鄢文静
张雨婷
谭子虎
杨琼
QIU Jing;YAN Wen-jing;ZHANG Yu-ting;TAN Zi-hu;YANG Qiong(College of Clinical Chinese Medicine,Hubei University of Chinese Medicine,Hubei Wuhan 430061,China)
出处
《安徽中医药大学学报》
2020年第1期60-64,共5页
Journal of Anhui University of Chinese Medicine
基金
中国中医科学院中医基础理论研究所自主选题“院所协同创新科研专项基金”(YZ-1852)
湖北省自然科学基金重点项目(2015CFA089)
湖北中医药大学研究生创新能力提升项目.
关键词
阿尔茨海默病
加减薯蓣丸
自噬
转录因子EB
Alzheimer’s disease
Modified Dioscorea opposita Thunb.Pill
Autophagy
Transcription factor EB
作者简介
邱静(1988-),女,博士研究生;通信作者:谭子虎(1964-),男,博士,主任医师,博士研究生导师,tanzihu2008@163.com
