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Aging and disease    2019, Vol. 10 Issue (6) : 1233-1245     DOI: 10.14336/AD.2018.1024
Orginal Article |
Autophagy Activation is Associated with Neuroprotection in Diabetes-associated Cognitive Decline
Yanqing Wu1,2, Libing Ye1, Yuan Yuan1, Ting Jiang1, Xin Guo1, Zhouguang Wang1, Ke Xu2, Zeping Xu1, Yanlong Liu1, Xingfeng Zhong3, Junmin Ye3, Hongyu Zhang1, Xiaokun Li1,*, Jian Xiao1,*
1Molecular Pharmacology Research Center, School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou, Zhejiang, China
2The Institute of Life Sciences, Wenzhou University, Wenzhou, Zhejiang, China
3Department of Anesthesia, The First Affiliated Hospital, Gannan Medical University, Jiangxi, China
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Autophagy is a lysosome-dependent cellular catabolic mechanism that mediates the turnover of dysfunctional organelles and aggregated proteins. It has a neuroprotective role on neurodegenerative diseases. Here, we hypothesized that autophagy may also have a neuroprotective role in diabetes-associated cognitive decline (DACD). In current study, we found that db/db mice display cognitive decline with inferior learning and memory function. The accumulation of β-amyloid1-42 (Aβ1-42), which is a characteristic of Alzheimer’s disease (AD), was markedly higher in the prefrontal cortex (PFC), cornu ammon1 (CA1), and dentate gyrus (DG) areas of the hippocampus in db/db mice. Moreover, BDNF and microtubule associated protein 2 (MAP2) levels were lower in the hippocampus of db/db mice. However, there was no noticeable differences in the level of apoptosis in the hippocampus between control (CON) mice and db/db mice. Markers of autophagy in the hippocampus were elevated in db/db mice. The expression levels of ATG5, ATG7, and LC3B were higher, and the level of P62 was lower. An autophagy inhibitor, 3-MA, and ATG7 siRNA significantly reversed the activation of autophagy in vitro, which was accompanied with a higher level of apoptosis. Taken together, our current study suggests that diabetes is associated with cognitive decline, and activation of autophagy has a neuroprotective role in DACD.

Keywords Diabetes-associated cognitive decline (DACD)      Autophagy      Hippocampus      β-amyloid      Apoptosis     
Corresponding Authors: Li Xiaokun,Xiao Jian   
About author: These authors contributed equally to this work.
Just Accepted Date: 12 November 2018   Issue Date: 16 November 2019
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Wu Yanqing
Ye Libing
Yuan Yuan
Jiang Ting
Guo Xin
Wang Zhouguang
Xu Ke
Xu Zeping
Liu Yanlong
Zhong Xingfeng
Ye Junmin
Zhang Hongyu
Li Xiaokun
Xiao Jian
Cite this article:   
Wu Yanqing,Ye Libing,Yuan Yuan, et al. Autophagy Activation is Associated with Neuroprotection in Diabetes-associated Cognitive Decline[J]. Aging and disease, 2019, 10(6): 1233-1245.
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Figure 1.  Diabetes resulted in cognitive decline with inferior learning and memory function. (A) The learning curve of training during six blocks in the Morris water maze test of CON mice and db/db mice. (B) Number of crossings over the original platform location in CON mice and db/db mice at 1 hr and 24 hr after training. (C) Representative swimming track of CON mice and db/db mice at 1 hr and 24 hr after training. (D) Swimming speed of CON mice and db/db mice. CON: control. **p < 0.01 vs. CON, ***p < 0.001 vs. CON, and N = 10.
Figure 2.  Diabetes induced Aβ1-42 accumulation and downregulated the expression of BDNF and MAP2. (A-B) The immunohistochemical staining of Aβ1-42 and BDNF in CA1, CA3, and PFC of the hippocampus. (C) The immunofluorescence staining of MAP2 in CA1, CA3, and PFC. CON: control, PFC: prefrontal cortex, CA1: cornu ammon1, and CA3: cornu ammon3Scale bar = 50 μm, and N = 4.
Figure 3.  The morphological structure and level of apoptosis in the hippocampus in diabetes. (A) H&E staining of CA1 and DG. (B) TUNEL staining of the hippocampus in CON mice and db/db mice. CON: control. Scale bar = 50 μm, and N = 4.
Figure 4.  The expression levels of LC3B and P62 in the hippocampus. (A) Western blot and quantitative analysis of LC3B and P62 expression. (B) Immunofluorescence staining of LC3B in CA1. CON: control. Scale bar = 50 µm and 10 µm. **p < 0.01 vs. CON, ***p <0.001 vs. CON, and N = 6.
Figure 5.  The expression levels of ATG5 and ATG7 in the hippocampus. (A) Western blot and quantitative analysis of ATG5 and ATG7. (B) Immunohistochemical staining of ATG5 and ATG7 in CA1 and CA3. CON: control. Scale bar = 50 µm. **p < 0.01 vs. CON, and N = 6.
Figure 6.  HG treatment increased the expression of autophagy markers in PC12 cells. PC12 cells were cultured in HG (30 mM) conditions for 12, 24, 36, and 48 hr. (A) Western blot analysis of ATG7, ATG5, P62, and LC3B expression. (B) Quantitative analysis of LC3B and P62. (C) Quantitative analysis of ATG5 and ATG7. (D) Immunofluorescence of LC3B in PC12 cells. CON: control, HG: high glucose. Scale bar = 50 μm, *P < 0.05 vs.CON, **P < 0.01vs. CON, ***P < 0.001 vs. CON, and N = 3.
Figure 7.  3-MA treatment inhibited HG-mediated autophagic activation, resulting in the induction of apoptosis. (A) Western blot analysis of ATG7, ATG5, P62, and LC3B expression. (B) Quantitative analysis of ATG5 and ATG7. (C) Quantitative analysis of LC3B and P62. (D) Immunofluorescence staining of LC3B in PC12 cells. (E) Cells were collected and stained with annexin V-FITC/propidium iodide and detected by flow cytometry. The lower right panel indicates the apoptotic cells. (F) The level of apoptosis in PC12 cells. CON: control, HG: high glucose. Scale bar = 50 μm, **P < 0.01 vs. CON, ***P < 0.001 vs. CON, #P < 0.05 vs. HG group, ##P < 0.01 vs. HG group, ###P < 0.001 vs. HG group, and N = 3.
Figure 8.  ATG7 siRNA treatment abolished the neuroprotective role of autophagy under HG conditions. (A) Western blot analysis of ATG7, ATG5, P62, and LC3B expression. (B) Quantitative analysis of ATG5 and ATG7. (C) Quantitative analysis of LC3B and P62. (D) Immunofluorescence staining of LC3B in PC12 cells. (E) The results of flow cytometry under different conditions. (F) The level of apoptosis in PC12 cells. CON: control, HG: high glucose. Scale bar = 50 μm. *P < 0.05 vs. CON, **P < 0.01 vs. CON, ***P < 0.001 vs. CON, #P < 0.05 vs. HG group, ##P < 0.01 vs. HG group, ###P < 0.001vs. HG group, and N = 3.
[1] Enomoto M, Yoshii H, Mita T, Sanke H, Yokota A, Yamashiro K, et al (2015). Relationship between dietary pattern and cognitive function in elderly patients with type 2 diabetes mellitus. J Int Med Res, 43: 506-17.
[2] Wang Y, Xu X, Feng C, Li Y, Ge X, Zong G, et al (2015). Patients with type 2 diabetes exhibit cognitive impairment with changes of metabolite concentration in the left hippocampus. Metab Brain Dis, 30: 1027-34.
[3] Mijnhout GS, Scheltens P, Diamant M, Biessels GJ, Wessels AM, Simsek S, et al (2006). Diabetic encephalopathy: a concept in need of a definition. Diabetologia, 49: 1447-8.
[4] Dominguez RO, Marschoff ER, Gonzalez SE, Repetto MG, Serra JA (2012). Type 2 diabetes and/or its treatment leads to less cognitive impairment in Alzheimer’s disease patients. Diabetes Res Clin Pr, 98: 68-74.
[5] Barbagallo M, Dominguez LJ (2014). Type 2 diabetes mellitus and Alzheimer’s disease. World J diabetes, 5: 889-93.
[6] Carvalho C, Machado N, Mota PC, Correia SC, Cardoso S, Santos RX, et al (2013). Type 2 Diabetic and Alzheimer’s Disease Mice Present Similar Behavioral, Cognitive, and Vascular Anomalies. J Alzheimers Dis, 35: 623-35.
[7] Arvanitakis Z, Wilson RS, Bienias JL, Evans DA, Bennett DA (2004). Diabetes mellitus and risk of Alzheimer disease and decline in cognitive function. Arch Neurol, 61: 661-6.
[8] Koekkoek PS, Ruis C, van den Donk M, Biessels GJ, Gorter KJ, Kappelle LJ, et al (2012). Intensive multifactorial treatment and cognitive functioning in screen-detected type 2 diabetes-The ADDITION-Netherlands study: A cluster-randomized trial. J Neurol Sci, 314: 71-7.
[9] Kariharan T, Nanayakkara G, Parameshwaran K, Bagasrawala I, Ahuja M, Abdel-Rahman E, et al (2015). Central activation of PPAR-gamma ameliorates diabetes induced cognitive dysfunction and improves BDNF expression. Neurobiol Aging, 36: 1451-61.
[10] Chen J, Liang L, Zhan L, Zhou Y, Zheng L, Sun X, et al (2014). ZiBuPiYin Recipe Protects db/db Mice from Diabetes-Associated Cognitive Decline through Improving Multiple Pathological Changes. Plos One, 9: e91680.
[11] Cuervo AM, Bergamini E, Brunk UT, Droge W, Ffrench M, Terman A (2005). Autophagy and aging-The importance of maintaining “clean” cells. Autophagy, 1: 131-40.
[12] Mizushima N, Levine B, Cuervo AM, Klionsky DJ (2008). Autophagy fights disease through cellular self-digestion. Nature, 451: 1069-75.
[13] Nikoletopoulou V, Papandreou M-E, Tavernarakis N (2015). Autophagy in the physiology and pathology of the central nervous system. Cell Death Differ, 22: 398-407.
[14] Yamamoto A, Yue Z (2014). Autophagy and Its Normal and Pathogenic States in the Brain. In: Hyman SE, editor. Annu Rev Neurosci, 55-78.
[15] Zhu X-C, Yu J-T, Jiang T, Tan L (2013). Autophagy Modulation for Alzheimer’s Disease Therapy. Mol Neurobiol, 48: 702-14.
[16] Xie Y, Kang R, Sun X, Zhong M, Huang J, Klionsky DJ, et al (2015). Posttranslational modification of autophagy-related proteins in macroautophagy. Autophagy, 11: 28-45.
[17] Tung Y-T, Wang B-J, Hu M-K, Hsu W-M, Lee H, Huang W-P, et al (2012). Autophagy: A double-edged sword in Alzheimer’s disease. J Biosciences, 37: 157-65.
[18] Katayama H, Kogure T, Mizushima N, Yoshimori T, Miyawaki A (2011). A Sensitive and Quantitative Technique for Detecting Autophagic Events Based on Lysosomal Delivery. Chem Biol, 18: 1042-52.
[19] Ichimura Y, Komatsu M (2010). Selective degradation of p62 by autophagy. Semin Immunopathol, 32: 431-6.
[20] Brownlee M (2001). Biochemistry and molecular cell biology and diabetic complications. Nature, 414: 813-20.
[21] Carlsson CM (2010). Type 2 Diabetes Mellitus, Dyslipidemia, and Alzheimer’s Disease. J Alzheimers Dis, 20: 711-22.
[22] Jiang B, Le L, Liu H, Xu L, He C, Hu K, Peng Y, et al (2016). Marein protects against methylglyoxal-induced apoptosis by activating the AMPK pathway in PC12 cells. Free Radical Res, 50: 1173-87.
[23] Martinez-Vicente M (2015). Autophagy in neurodegenerative diseases: From pathogenic dysfunction to therapeutic modulation. Semin Cell Dev Biol, 40: 115-26.
[24] Kochergin IA, Zakharova MN (2016). The role of autophagy in neurodegenerative diseases. Neurochem J, 10: 7-18.
[25] Beilina A, Cookson MR (2016). Genes associated with Parkinson’s disease: regulation of autophagy and beyond. J Neurochem, 139: 91-107.
[26] Navone F, Genevini P, Borgese N (2015). Autophagy and Neurodegeneration: Insights from a Cultured Cell Model of ALS. Cells, 4: 354-86.
[27] Rui Y-N, Xu Z, Patel B, Cuervo AM, Zhang S (2015). HTT/Huntingtin in selective autophagy and Huntington disease: A foe or a friend within? Autophagy, 11: 858-60.
[28] Singh AK, Kashyap MP, Tripathi VK, Singh S, Garg G, Rizvi SI (2017). Neuroprotection Through Rapamycin-Induced Activation of Autophagy and PI3K/Akt1/mTOR/CREB Signaling Against Amyloid-beta-Induced Oxidative Stress, Synaptic/Neurotransmission Dysfunction, and Neurodegeneration in Adult Rats. Mol Neurobiol, 54: 5815-28.
[29] Li Z, Zhang W, Sima AAF (2003). C-peptide enhances insulin-mediated cell growth and protection against high glucose-induced apoptosis in SH-SY5Y cells. Diabetes-Metab Res, 19: 375-85.
[30] Sale P, De Pandis MF, Vimercati SL, Sova I, Foti C, Tenore N, et al (2013). The relation between Parkinson’s disease and ageing Comparison of the gait patterns of young Parkinson’s disease subjects with healthy elderly subjects. Eur J Phys Rehab Med, 49: 161-7.
[31] Hayden MS, Ghosh S (2004). Signaling to NF-kappa B. Genes Dev, 18: 2195-224.
[32] Mario UD, Morano S, Valle E, Pozzessere G(1995). Electrophysiological alteration of the central nervous system in diabetes mellitus. Diab Metab Rev, 11:259-72.
[33] Bliss TV, Collingriage GL(1993). A synapfic model of memory: long-term potentiation in the hippocampus. Nature, 361(1):3-9.
[34] Yang Z, Klionsky DJ (2010). Eaten alive: a history of macroautophagy. Nat Cell Biol, 12: 814-22.
[35] Mizushima N, Komatsu M (2011). Autophagy: Renovation of Cells and Tissues. Cell, 147: 728-41.
[36] Wong E, Cuervo AM (2010). Autophagy gone awry in neurodegenerative diseases. Nat Neurosci, 13: 805-11.
[37] Madeo F, Eisenberg T, Kroemer G (2009). Autophagy for the avoidance of neurodegeneration. Gene Dev, 23: 2253-9.
[38] Ling D, Salvaterra PM (2009). A central role for autophagy in Alzheimer-type neurodegeneration. Autophagy, 5: 738-40.
[39] Matsuoka Y, Jouroukhin Y, Gray AJ, Ma L, Hirata-Fukae C, Li H-F, et al (2008). A neuronal microtubule-interacting agent, NAPVSIPQ, reduces tau pathology and enhances cognitive function in a mouse model of Alzheimer’s disease. J Pharmacol Exp Ther, 325: 146-53.
[40] Allen DA, Yaqoob MM, Harwood SM (2005). Mechanisms of high glucose-induced apoptosis and its relationship to diabetic complications. J Nutr Biochem, 16: 705-13.
[41] Susztak K, Raff AC, Schiffer M, Bottinger EP (2006). Glucose-induced reactive oxygen species cause apoptosis of podocytes and podocyte depletion at the onset of diabetic nephropathy. Diabetes, 55: 225-33.
[42] Lin JH, Walter P, Yen TSB (2008). Endoplasmic reticulum stress in disease pathogenesis. Annu Rev Pathol-Mech, 399-425.
[43] Eizirik DL, Cardozo AK, Cnop M (2008). The role for endoplasmic reticulum stress in diabetes mellitus. Endocr Rev, 29: 42-61.
[44] Chiang C-K, Wang C-C, Lu T-F, Huang K-H, Sheu M-L, Liu S-H, et al (2016). Involvement of Endoplasmic Reticulum Stress, Autophagy, and Apoptosis in Advanced Glycation End Products-Induced Glomerular Mesangial Cell Injury. Sci Rep, 6: 34167.
[45] Sims-Robinson C, Zhao S, Hur J, Feldman EL (2012). Central nervous system endoplasmic reticulum stress in a murine model of type 2 diabetes. Diabetologia, 55: 2276-84.
[47] Perea G, Araque A (2005). Glial calcium signaling and neuron-glia communication. Cell Calcium, 38: 375-82.
[48] Duarte JMN, Agostinho PM, Carvalho RA, Cunha RA (2012). Caffeine Consumption Prevents Diabetes-Induced Memory Impairment and Synaptotoxicity in the Hippocampus of NONcZNO10/LTJ Mice. Plos One, 7: e21899.
[49] Zheng Y, Yang Y, Dong B, Zheng H, Lin X, Du Y, et al (2016). Metabonomic profiles delineate potential role of glutamate-glutamine cycle in db/db mice with diabetes-associated cognitive decline. Mol Brain, 9: 40.
[50] Galluzzi L, Pietrocola F, Levine B, Kroemer G (2014). Metabolic Control of Autophagy. Cell, 159: 1263-76.
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