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Hydroxytyrosol improves mitochondrial function and reduces oxidative stress in the brain of db/db mice: role of AMP-activated protein kinase activation

  • Adi Zheng (a1), Hao Li (a1), Jie Xu (a1), Ke Cao (a2), Hua Li (a1), Wenjun Pu (a1), Ziqi Yang (a1), Yunhua Peng (a2), Jiangang Long (a1), Jiankang Liu (a1) and Zhihui Feng (a2)...


Hydroxytyrosol (HT) is a major polyphenolic compound found in olive oil with reported anti-cancer and anti-inflammatory activities. However, the neuroprotective effect of HT on type 2 diabetes remains unknown. In the present study, db/db mice and SH-SY-5Y neuroblastoma cells were used to evaluate the neuroprotective effects of HT. After 8 weeks of HT administration at doses of 10 and 50 mg/kg, expression levels of the mitochondrial respiratory chain complexes I/II/IV and the activity of complex I were significantly elevated in the brain of db/db mice. Likewise, targets of the antioxidative transcription factor nuclear factor erythroid 2 related factor 2 including p62 (sequestosome-1), haeme oxygenase 1 (HO-1), and superoxide dismutases 1 and 2 increased, and protein oxidation significantly decreased. HT treatment was also found to activate AMP-activated protein kinase (AMPK), sirtuin 1 and PPARγ coactivator-1α, which constitute an energy-sensing protein network known to regulate mitochondrial function and oxidative stress responses. Meanwhile, neuronal survival indicated by neuron marker expression levels including activity-regulated cytoskeleton-associated protein, N-methyl-d-aspartate receptor and nerve growth factor was significantly improved by HT administration. Additionally, in a high glucose-induced neuronal cell damage model, HT effectively increased mitochondrial complex IV and HO-1 expression through activating AMPK pathway, followed by the prevention of high glucose-induced production of reactive oxygen species and declines of cell viability and VO2 capacity. Our observations suggest that HT improves mitochondrial function and reduces oxidative stress potentially through activation of the AMPK pathway in the brain of db/db mice.

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Corresponding author

* Corresponding author: J. Liu, fax +86 29 82665849, email; Z. Feng, fax +86 29 82665849, email


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1 Whiting, DR, Guariguata, L, Weil, C, et al. (2011) IDF diabetes atlas: global estimates of the prevalence of diabetes for 2011 and 2030. Diabetes Res Clin Pract 94, 311321.
2 Stumvoll, M, Goldstein, BJ & van Haeften, TW (2005) Type 2 diabetes: principles of pathogenesis and therapy. Lancet 365, 13331346.
3 Manschot, SM, Brands, AM, van der Grond, J, et al. (2006) Brain magnetic resonance imaging correlates of impaired cognition in patients with type 2 diabetes. Diabetes 55, 11061113.
4 Gold, SM, Dziobek, I, Sweat, V, et al. (2007) Hippocampal damage and memory impairments as possible early brain complications of type 2 diabetes. Diabetologia 50, 711719.
5 Qiu, C, Sigurdsson, S, Zhang, Q, et al. (2014) Diabetes, markers of brain pathology, and cognitive cognition: the AGES-Reykjavik Study. Ann Neurol 75, 138146.
6 Novak, V, Zhao, P, Manor, B, et al. (2011) Adhesion molecules, altered vasoreactivity, and brain atrophy in type 2 diabetes. Diabetes Care 34, 24382441.
7 Moran, C, Phan, TG, Chen, J, et al. (2013) Brain atrophy in type 2 diabetes: regional distribution and influence on cognition. Diabetes Care 36, 40364042.
8 Kumar, A, Haroon, E, Darwin, C, et al. (2008) Gray matter prefrontal changes in type 2 diabetes detected using MRI. J Magn Reson Imaging 27, 1419.
9 Brundel, M, van den Heuvel, M, de Bresser, J, et al. (2010) Cerebral cortical thickness in patients with type 2 diabetes. J Neurol Sci 299, 126130.
10 Zhao, B, Pan, BS, Shen, SW, et al. (2013) Diabetes-induced central neuritic dystrophy and cognitive deficits are associated with the formation of oligomeric reticulon-3 via oxidative stress. J Biol Chem 288, 1559015599.
11 Correia, SC, Santos, RX, Carvalho, C, et al. (2012) Insulin signaling, glucose metabolism and mitochondria: major players in Alzheimer's disease and diabetes interrelation. Brain Res 1441, 6478.
12 Milone, M (2012) Mitochondria, diabetes, and Alzheimer's disease. Diabetes 61, 991992.
13 Ceriello, A (2003) New insights on oxidative stress and diabetic complications may lead to a “causal” antioxidant therapy. Diabetes Care 26, 15891596.
14 Vincent, AM, Russell, JW, Sullivan, KA, et al. (2007) SOD2 protects neurons from injury in cell culture and animal models of diabetic neuropathy. Exp Neurol 208, 216227.
15 Valente, T, Gella, A, Fernandez-Busquets, X, et al. (2010) Immunohistochemical analysis of human brain suggests pathological synergism of Alzheimer's disease and diabetes mellitus. Neurobiol Dis 37, 6776.
16 Teshima, Y, Takahashi, N, Nishio, S, et al. (2014) Production of reactive oxygen species in the diabetic heart. Roles of mitochondria and NADPH oxidase. Circ J 78, 300306.
17 Moreira, PI, Santos, MS, Seica, R, et al. (2007) Brain mitochondrial dysfunction as a link between Alzheimer's disease and diabetes. J Neurol Sci 257, 206214.
18 Visioli, F, Bellomo, G & Galli, C (1998) Free radical-scavenging properties of olive oil polyphenols. Biochem Biophys Res Commun 247, 6064.
19 Stupans, I, Kirlich, A, Tuck, KL, et al. (2002) Comparison of radical scavenging effect, inhibition of microsomal oxygen free radical generation, and serum lipoprotein oxidation of several natural antioxidants. J Agric Food Chem 50, 24642469.
20 Zhu, L, Liu, Z, Feng, Z, et al. (2010) Hydroxytyrosol protects against oxidative damage by simultaneous activation of mitochondrial biogenesis and phase II detoxifying enzyme systems in retinal pigment epithelial cells. J Nutr Biochem 21, 10891098.
21 Zou, X, Feng, Z, Li, Y, et al. (2012) Stimulation of GSH synthesis to prevent oxidative stress-induced apoptosis by hydroxytyrosol in human retinal pigment epithelial cells: activation of Nrf2 and JNK-p62/SQSTM1 pathways. J Nutr Biochem 23, 9941006.
22 Liu, Z, Sun, L, Zhu, L, et al. (2007) Hydroxytyrosol protects retinal pigment epithelial cells from acrolein-induced oxidative stress and mitochondrial dysfunction. J Neurochem 103, 26902700.
23 Hao, J, Shen, W, Yu, G, et al. (2010) Hydroxytyrosol promotes mitochondrial biogenesis and mitochondrial function in 3T3-L1 adipocytes. J Nutr Biochem 21, 634644.
24 Wu, YT, Lin, LC & Tsai, TH (2009) Measurement of free hydroxytyrosol in microdialysates from blood and brain of anesthetized rats by liquid chromatography with fluorescence detection. J Chromatogr A 1216, 35013507.
25 Cao, K, Xu, J, Zou, X, et al. (2014) Hydroxytyrosol prevents diet-induced metabolic syndrome and attenuates mitochondrial abnormalities in obese mice. Free Radic Biol Med 67, 396407.
26 Komatsu, M, Kurokawa, H, Waguri, S, et al. (2010) The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1. Nat Cell Biol 12, 213223.
27 Lira, VA, Brown, DL, Lira, AK, et al. (2010) Nitric oxide and AMPK cooperatively regulate PGC-1 in skeletal muscle cells. J Physiol 588, 35513566.
28 Chau, MD, Gao, J, Yang, Q, et al. (2010) Fibroblast growth factor 21 regulates energy metabolism by activating the AMPK–SIRT1–PGC-1alpha pathway. Proc Natl Acad Sci U S A 107, 1255312558.
29 Mo, C, Wang, L, Zhang, J, et al (2014) The crosstalk between Nrf2 and AMPK signal pathways is important for the anti-inflammatory effect of berberine in LPS-stimulated macrophages and endotoxin-shocked mice. Antioxid Redox Signal 20, 574588.
30 Shi, TJ, Zhang, MD, Zeberg, H, et al. (2013) Coenzyme Q10 prevents peripheral neuropathy and attenuates neuron loss in the db-/db-mouse, a type 2 diabetes model. Proc Natl Acad Sci U S A 110, 690695.
31 Kapoor, R & Kakkar, P (2012) Protective role of morin, a flavonoid, against high glucose induced oxidative stress mediated apoptosis in primary rat hepatocytes. PLOS ONE 7, e41663.
32 Hsu, YY, Tseng, YT & Lo, YC (2013) Berberine, a natural antidiabetes drug, attenuates glucose neurotoxicity and promotes Nrf2-related neurite outgrowth. Toxicol Appl Pharmacol 272, 787796.
33 Li, Y, Xu, S, Zhang, Q, et al. (2014) Cytotoxicity study on SHSY5Y cells cultured at high glucose levels and treated with bupivacaine. Mol Med Rep 9, 515520.
34 Li, L, Ye, XP, Lu, AZ, et al. (2013) Hyperglycemia magnifies bupivacaine-induced cell apoptosis triggered by mitochondria dysfunction and endoplasmic reticulum stress. J Neurosci Res 91, 786798.
35 Russell, JW, Golovoy, D, Vincent, AM, et al. (2002) High glucose-induced oxidative stress and mitochondrial dysfunction in neurons. FASEB J 16, 17381748.
36 Lourida, I, Soni, M, Thompson-Coon, J, et al. (2013) Mediterranean diet, cognitive function, and dementia: a systematic review. Epidemiology 24, 479489.
37 Valls-Pedret, C, Lamuela-Raventos, RM, Medina-Remon, A, et al. (2012) Polyphenol-rich foods in the Mediterranean diet are associated with better cognitive function in elderly subjects at high cardiovascular risk. J Alzheimer's Dis 29, 773782.
38 Li, XL, Aou, S, Oomura, Y, et al. (2002) Impairment of long-term potentiation and spatial memory in leptin receptor-deficient rodents. Neuroscience 113, 607615.
39 Ernst, A, Sharma, A, Elased, K, et al. (2013) Diabetic db/db mice exhibit central nervous system and peripheral molecular alterations as seen in neurological disorders. Transl Psychiatry 3, e263.
40 Guzowski, JF, Lyford, GL, Stevenson, GD, et al. (2000) Inhibition of activity-dependent arc protein expression in the rat hippocampus impairs the maintenance of long-term potentiation and the consolidation of long-term memory. J Neurosci 20, 39934001.
41 Mateos, L, Akterin, S, Gil-Bea, FJ, et al. (2009) Activity-regulated cytoskeleton-associated protein in rodent brain is down-regulated by high fat diet in vivo and by 27-hydroxycholesterol in vitro . Brain Pathol 19, 6980.
42 Steward, O & Worley, PF (2001) Selective targeting of newly synthesized Arc mRNA to active synapses requires NMDA receptor activation. Neuron 30, 227240.
43 Unger, J, Klitzsch, T, Pera, S, et al. (1998) Nerve growth factor (NGF) and diabetic neuropathy in the rat: morphological investigations of the sural nerve, dorsal root ganglion, and spinal cord. Exp Neurol 153, 2334.
44 Muangman, P, Muffley, LA, Anthony, JP, et al. (2004) Nerve growth factor accelerates wound healing in diabetic mice. Wound Repair Regen 12, 4452.
45 Amato, S & Man, H-Y (2011) Bioenergy sensing in the brain: the role of AMP-activated protein kinase in neuronal metabolism, development and neurological diseases. Cell Cycle 10, 34523460.
46 Ronnett, GV, Ramamurthy, S, Kleman, AM, et al. (2009) AMPK in the brain: its roles in energy balance and neuroprotection. J Neurochem 109, 1723.
47 Chowdhury, SKR, Smith, DR, Saleh, A, et al. (2012) Impaired adenosine monophosphate-activated protein kinase signalling in dorsal root ganglia neurons is linked to mitochondrial dysfunction and peripheral neuropathy in diabetes. Brain 135, 17511766.
48 Kitada, M, Kume, S, Imaizumi, N, et al. (2011) Resveratrol improves oxidative stress and protects against diabetic nephropathy through normalization of Mn-SOD dysfunction in AMPK/SIRT1-independent pathway. Diabetes 60, 634643.
49 Zrelli, H, Matsuoka, M, Kitazaki, S, et al. (2011) Hydroxytyrosol reduces intracellular reactive oxygen species levels in vascular endothelial cells by upregulating catalase expression through the AMPK–FOXO3a pathway. Eur J Pharmacol 660, 275282.
50 Visioli, F, Caruso, D, Plasmati, E, et al. (2001) Hydroxytyrosol, as a component of olive mill waste water, is dose-dependently absorbed and increases the antioxidant capacity of rat plasma. Free Radic Res 34, 301305.
51 Ruiz-Gutierrez, V, Juan, ME, Cert, A, et al. (2000) Determination of hydroxytyrosol in plasma by HPLC. Anal Chem 72, 44584461.
52 Tuck, KL & Hayball, PJ (2002) Major phenolic compounds in olive oil: metabolism and health effects. J Nutr Biochem 13, 636644.
53 Vazquez-Velasco, M, Esperanza Diaz, L, Lucas, R, et al. (2011) Effects of hydroxytyrosol-enriched sunflower oil consumption on CVD risk factors. Br J Nutr 105, 14481452.



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