电针“百会”“涌泉”对4月龄APP/PS1双转基因小鼠海马LC3和Aβ影响的研究
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摘要
目的:探讨电针对4月龄APP/PS1双转基因小鼠海马LC3和Aβ的影响。方法:将16只4月龄雄性APP/PS1双转基因小鼠随机分为电针组和模型组;以8只4月龄雄性C57BL/6小鼠作为正常组。电针组电针"百会""涌泉"穴,正常组、模型组以与电针组相同的方法束缚,干预均隔日一次,6周后取其脑组织。免疫组化法观察小鼠皮质和海马内LC3的表达情况;免疫荧光双标法观察小鼠海马内LC3和Aβ的共表达情况;Western blot(WB)法检测小鼠海马内LC3Ⅰ、LC3Ⅱ的表达水平。结果:免疫组化结果显示,在小鼠海马和皮质中,LC3主要表达于神经元的胞质和轴突。免疫荧光双标法结果显示,LC3和Aβ存在共表达。WB结果显示,正常组LC3Ⅱ/LC3Ⅰ比值大于模型组,差异有统计学意义(P<0.01);电针组LC3Ⅱ/LC3Ⅰ比值大于模型组,但差异无统计学意义(P>0.05,P=0.19)。结论:5月龄时APP/PS1双转基因小鼠海马内自噬处于抑制状态;电针干预对4月龄APP/PS1小鼠海马内的自噬功能是否存在促进作用,需要进一步实验证实。
Objective: To observe the effects of electroacupuncture on the level of LC3 in brain in APP/PS1 transgenic mice. Methods: A total of 16 male APP/PS1 transgenic mice were divided into electroacupuncture group and model group randomly. There were 8 male C57 BL/6 mice selected as normal control group. The expression of LC3 in brain was ob?served by immunohistochemistry after 6 weeks of electroacupuncture at Baihui(DU 20) and Yongquan(KI 1). Co-expres?sion of LC3 and Aβ in brain was observed by immunofluorescence double labeling. The level of LC3Ⅰ、LC3Ⅱ in hippo?campus was detected by Western blot(WB). Results: Immunohistochemistry results showed that expression of LC3 was in neurons in hippocampus and cortex. The results of immun of luorescence double labeling showed that there was co-ex?pression of LC3 and Aβ. WB results showed that the level of LC3Ⅱ/LC3Ⅰin the model group(P<0.01) and electroacu?puncture group(P<0.05) both were lower than the level in control group. Compared with the model group, the level of LC3Ⅱ/LC3Ⅰin the electroacupuncture group was increased(P > 0.05, P = 0.19). Conclusion: Autophagy in the hippo?campus of 5-month APP/PS1 double transgenic mice was inhibited; Whether electroacupuncture intervention has an ef?fect on the autophagy level in hippocampus of 4 months old APP/PS1 mice needs confirmation futher.
Objective: To observe the effects of electroacupuncture on the level of LC3 in brain in APP/PS1 transgenic mice. Methods: A total of 16 male APP/PS1 transgenic mice were divided into electroacupuncture group and model group randomly. There were 8 male C57 BL/6 mice selected as normal control group. The expression of LC3 in brain was ob?served by immunohistochemistry after 6 weeks of electroacupuncture at Baihui(DU 20) and Yongquan(KI 1). Co-expres?sion of LC3 and Aβ in brain was observed by immunofluorescence double labeling. The level of LC3Ⅰ、LC3Ⅱ in hippo?campus was detected by Western blot(WB). Results: Immunohistochemistry results showed that expression of LC3 was in neurons in hippocampus and cortex. The results of immun of luorescence double labeling showed that there was co-ex?pression of LC3 and Aβ. WB results showed that the level of LC3Ⅱ/LC3Ⅰin the model group(P<0.01) and electroacu?puncture group(P<0.05) both were lower than the level in control group. Compared with the model group, the level of LC3Ⅱ/LC3Ⅰin the electroacupuncture group was increased(P > 0.05, P = 0.19). Conclusion: Autophagy in the hippo?campus of 5-month APP/PS1 double transgenic mice was inhibited; Whether electroacupuncture intervention has an ef?fect on the autophagy level in hippocampus of 4 months old APP/PS1 mice needs confirmation futher.
引文
1王行健,郑玮.阿尔兹海默病与自噬.中国老年学杂志, 2017, 8(37):3876-3881.
2 Boland B, Kumar A, Nixon RA, et al. Autophagy induction and autophagosome clearance in neurons:relationship to autophagic pathology in Alzheimer’s disease. J Neurosci, 2008, 28(27):6926-6937.
3 Nixon R A, Yang D S. Autophagy failure in Alzheimer's disease--locating the primary defect. Neurobiol Dis, 2011, 43(1):38-45.
4 LiangJH, JiaJP. Dysfunctional autophagyinAlzheimer'sdisease:pathogenic roles and therapeutic implications. Neurosci Bull, 2014, 30(2):308-316.
5 Saadipour K, Yang M, Lim Y, et al. Amyloid beta1-42(Aβ42)up-regulates the expression of sortilin via the p75NTR/RhoA signaling pathway. J Neurochem, 2013, 127(2):152-162.
6 Zhang W, Bai M, Hao J, et al. Early memory deficits precede plaque deposition in APPswe/PS1dE9 mice:Involvement of oxidative stress and cholinergic dysfunction. Free Radic Biol Med, 2012, 52(8):1443-1452.
7 高堂珂,薛卫国. APP/PS1小鼠脑内Aβ水平与认知行为学增龄性变化的研究概述.中风与神经疾病杂志, 2015, 32(2):189-190.
8 Klionsky D J, Abdelmohsen K, Abe A, et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy, 2016, 12(1):1-222.
9 陈吉祥,吴羽楠,陈立典,等.电针百会穴对APP/PS1双转基因小鼠学习记忆及脑源性神经营养因子表达的影响.中国康复理论与实践, 2015, 21(6):642-647.
10 邵千枫,李昱颉,李志刚,等.“通督启神”法两种电针对APP/PS1小鼠额叶皮层小胶质细胞TNF-α表达的影响.世界科学技术—中医药现代化, 2016, 18(8):1327-1333.
11 Guo H D, Zhu J, Tian J X, et al. Electroacupuncture improves memory and protects neurons by regulation of the autophagy pathway in a rat model of Alzheimer's disease. Acupunct Med, 2016, 34(6):449-456.
12 薛卫国,张忠,许红,等.电针对APP转基因鼠神经元自噬途径的影响.针刺研究, 2014, 39(4):272-277.
13 李芙,姜玉莹,薛卫国,等.针刺百会、涌泉穴治疗老年痴呆病理论初步探讨.辽宁中医药大学学报, 2014, 16(10):143-145.
14 曹瑾,李昱颉,李志刚,等.督脉论治阿尔兹海默病及其机理研究.世界科学技术—中医药现代化, 2016, 18(8):1322-1326.
15 余梦瑶,许晓燕,江南,等.阿尔茨海默症痰湿蕴结与β-淀粉样肽聚集及自噬相关性探讨.中华中医药杂志, 2017, 31(9):3632-3635.
16 Hardy J, Selkoe D J. The amyloid hypothesis of Alzheimer's disease:progress and problems on the road to therapeutics. Science, 2002, 297(5580):353-356.
17 Yao Z G, Zhang L, Liang L, et al. The effect of PN-1 , a Traditional Chinese Prescription, on the Learning and Memory in a Transgenic Mouse Model of Alzheimer's Disease. Evid Based Complement Alternat Med, 2013, 2013:518421.
18 Salloway S, Sperling R, Fox NC, et al. Two phase 3 trials of bapineuzumab in mild-to-moderate Alzheimer's disease. N Engl J Med, 2014, 370(4):322-333.
19 Holtzman D M, Morris J C, Goate A M. Alzheimer’s disease:the challenge of the second century. Sci Transl Med, 2011, 3(77):1-35.
20 Eric K, Marc M, Bart D S. The amyloid cascade hypothesis for Alzheimer disease:an appraisal for the development of therapeutics. Nat Rev Drug Discov, 2011, 10(9):698-712.
21 Yu W H, Kumar A, Peterhoff C, et al. Autophagic vacuoles are enriched in amyloid precursor protein-secretase activities:implications for betaamyloid peptide over-production and localization in Alzheimer's disease. Int J Biochem Cell Biol, 2004, 36(12):2531-2540.
22 Cataldo A M, Petanceska S, Terio N B, et al. Abeta localization in abnormal endosomes:association with earliest Abeta elevations in AD and Down syndrome. Neurobiol Aging, 2004, 25(10):1263-1272.
23 Ling D, Magallanes M, Salvaterra P M. Accumulation of amyloid-like Aβ1-42 in AEL(autophagy-endosomal-lysosomal)vesicles:potential implications for plaque biogenesis. ASN Neuro, 2014, 6(2):95-109.
24 李芙,王鑫,薛卫国,等.电针“百会”“涌泉”对APP/PS1双转基因小鼠海马β淀粉样蛋白及低密度脂蛋白受体相关蛋白-1水平的影响.针刺研究, 2015, 40(1):30-55.
2 Boland B, Kumar A, Nixon RA, et al. Autophagy induction and autophagosome clearance in neurons:relationship to autophagic pathology in Alzheimer’s disease. J Neurosci, 2008, 28(27):6926-6937.
3 Nixon R A, Yang D S. Autophagy failure in Alzheimer's disease--locating the primary defect. Neurobiol Dis, 2011, 43(1):38-45.
4 LiangJH, JiaJP. Dysfunctional autophagyinAlzheimer'sdisease:pathogenic roles and therapeutic implications. Neurosci Bull, 2014, 30(2):308-316.
5 Saadipour K, Yang M, Lim Y, et al. Amyloid beta1-42(Aβ42)up-regulates the expression of sortilin via the p75NTR/RhoA signaling pathway. J Neurochem, 2013, 127(2):152-162.
6 Zhang W, Bai M, Hao J, et al. Early memory deficits precede plaque deposition in APPswe/PS1dE9 mice:Involvement of oxidative stress and cholinergic dysfunction. Free Radic Biol Med, 2012, 52(8):1443-1452.
7 高堂珂,薛卫国. APP/PS1小鼠脑内Aβ水平与认知行为学增龄性变化的研究概述.中风与神经疾病杂志, 2015, 32(2):189-190.
8 Klionsky D J, Abdelmohsen K, Abe A, et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy, 2016, 12(1):1-222.
9 陈吉祥,吴羽楠,陈立典,等.电针百会穴对APP/PS1双转基因小鼠学习记忆及脑源性神经营养因子表达的影响.中国康复理论与实践, 2015, 21(6):642-647.
10 邵千枫,李昱颉,李志刚,等.“通督启神”法两种电针对APP/PS1小鼠额叶皮层小胶质细胞TNF-α表达的影响.世界科学技术—中医药现代化, 2016, 18(8):1327-1333.
11 Guo H D, Zhu J, Tian J X, et al. Electroacupuncture improves memory and protects neurons by regulation of the autophagy pathway in a rat model of Alzheimer's disease. Acupunct Med, 2016, 34(6):449-456.
12 薛卫国,张忠,许红,等.电针对APP转基因鼠神经元自噬途径的影响.针刺研究, 2014, 39(4):272-277.
13 李芙,姜玉莹,薛卫国,等.针刺百会、涌泉穴治疗老年痴呆病理论初步探讨.辽宁中医药大学学报, 2014, 16(10):143-145.
14 曹瑾,李昱颉,李志刚,等.督脉论治阿尔兹海默病及其机理研究.世界科学技术—中医药现代化, 2016, 18(8):1322-1326.
15 余梦瑶,许晓燕,江南,等.阿尔茨海默症痰湿蕴结与β-淀粉样肽聚集及自噬相关性探讨.中华中医药杂志, 2017, 31(9):3632-3635.
16 Hardy J, Selkoe D J. The amyloid hypothesis of Alzheimer's disease:progress and problems on the road to therapeutics. Science, 2002, 297(5580):353-356.
17 Yao Z G, Zhang L, Liang L, et al. The effect of PN-1 , a Traditional Chinese Prescription, on the Learning and Memory in a Transgenic Mouse Model of Alzheimer's Disease. Evid Based Complement Alternat Med, 2013, 2013:518421.
18 Salloway S, Sperling R, Fox NC, et al. Two phase 3 trials of bapineuzumab in mild-to-moderate Alzheimer's disease. N Engl J Med, 2014, 370(4):322-333.
19 Holtzman D M, Morris J C, Goate A M. Alzheimer’s disease:the challenge of the second century. Sci Transl Med, 2011, 3(77):1-35.
20 Eric K, Marc M, Bart D S. The amyloid cascade hypothesis for Alzheimer disease:an appraisal for the development of therapeutics. Nat Rev Drug Discov, 2011, 10(9):698-712.
21 Yu W H, Kumar A, Peterhoff C, et al. Autophagic vacuoles are enriched in amyloid precursor protein-secretase activities:implications for betaamyloid peptide over-production and localization in Alzheimer's disease. Int J Biochem Cell Biol, 2004, 36(12):2531-2540.
22 Cataldo A M, Petanceska S, Terio N B, et al. Abeta localization in abnormal endosomes:association with earliest Abeta elevations in AD and Down syndrome. Neurobiol Aging, 2004, 25(10):1263-1272.
23 Ling D, Magallanes M, Salvaterra P M. Accumulation of amyloid-like Aβ1-42 in AEL(autophagy-endosomal-lysosomal)vesicles:potential implications for plaque biogenesis. ASN Neuro, 2014, 6(2):95-109.
24 李芙,王鑫,薛卫国,等.电针“百会”“涌泉”对APP/PS1双转基因小鼠海马β淀粉样蛋白及低密度脂蛋白受体相关蛋白-1水平的影响.针刺研究, 2015, 40(1):30-55.