Oxidative stress has been implicated in the pathophysiology of cerebral stroke. As NADPH oxidases (NOXs) play major roles in the regulation of oxidative stress, we hypothesized that reduction of NOX activity by depletion of p22phox, an essential subunit of NOX complexes, would prevent cerebral stroke. To investigate this, we used the stroke-prone spontaneously hypertensive rat (SHRSP) and the p22phox-deleted congenic SHRSP. Although p22phox depletion reduced blood pressure under salt loading, it did not ameliorate oxidative stress or reduce the incidence of salt-induced stroke in SHRSPs. Additional pharmacological reduction of oxidative stress using antioxidant reagents with different mechanisms of action was necessary to prevent stroke, indicating that NOX was not the major target in salt-induced stroke in SHRSPs. On the other hand, oxidative stress measured based on urinary isoprostane levels showed significant correlations with blood pressure, stroke latency and urinary protein excretion under salt loading, suggesting an important role of oxidative stress per se in hypertension and hypertensive organ damage. Overall, our results imply that oxidative stress from multiple sources influences stroke susceptibility and other hypertensive disorders in salt-loaded SHRSPs.
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Li W, Yang S. Targeting oxidative stress for the treatment of ischemic stroke: Upstream and downstream therapeutic strategies. Brain Circ. 2016;2:153.
Kleinschnitz C, Grund H, Wingler K, Armitage ME, Jones E, Mittal M, et al. Post-stroke inhibition of induced NADPH Oxidase type 4 prevents oxidative stress and neurodegeneration. PLoS Biol. 2010;8:e1000479.
Domínguez C, Delgado P, Vilches A, Martín-Gallán P, Ribó M, Santamarina E, et al. Oxidative stress after thrombolysis-induced reperfusion in human stroke. Stroke. 2010;41:653–60.
Moon GJ, Shin DH, Im DS, Bang OY, Nam HS, Lee JH, et al. Identification of oxidized serum albumin in the cerebrospinal fluid of ischaemic stroke patients. Eur J Neurol. 2011;18:1151–8.
Zhang L, Wu J, Duan X, Tian X, Shen H, Sun Q, et al. NADPH oxidase: a potential target for treatment of stroke. Oxid Med Cell Longev. 2016;2016:1–9.
Lipton P. Ischemic cell death in brain neurons. Physiol Rev. 1999;79:1431–568.
Zahid HM, Ferdaus MZ, Ohara H, Isomura M, Nabika T. Effect of p22phox depletion on sympathetic regulation of blood pressure in SHRSP: evaluation in a new congenic strain. Sci Rep. 2016;6:36739.
Shirley R, Ord E, Work L. Oxidative stress and the use of antioxidants in stroke. Antioxidants. 2014;3:472–501.
Watts LT, Lloyd R, Garling RJ, Duong T. Stroke neuroprotection: targeting mitochondria. Brain Sci. 2013;3:540–60.
Di Meo S, Reed TT, Venditti P, Victor VM. Role of ROS and RNS sources in physiological and pathological conditions. Oxid Med Cell Longev. 2016;2016:1245049.
Nomura J, Busso N, Ives A, Matsui C, Tsujimoto S, Shirakura T, et al. Xanthine oxidase inhibition by febuxostat attenuates experimental atherosclerosis in mice. Sci Rep. 2014;4:4554.
Touyz RM, Schiffrin EL. Reactive oxygen species in vascular biology: Implications in hypertension. Histochem Cell Biol. 2004;122:339–52.
Guzik TJ, Sadowski J, Guzik B, Jopek A, Kapelak B, Przybylowski P, et al. Coronary artery superoxide production and nox isoform expression in human coronary artery disease. Arterioscler Thromb Vasc Biol. 2006;26:333–9.
Bedard K, Krause K-H. The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiol Rev. 2007;87:245–313.
Kawahara T, Ritsick D, Cheng G, Lambeth JD. Point mutations in the proline-rich region of p22phox are dominant inhibitors of Nox1- and Nox2-dependent reactive oxygen generation. J Biol Chem. 2005;280:31859–69.
Chen F, Haigh S, Barman S, Fulton DJR. From form to function: the role of Nox4 in the cardiovascular system. Front Physiol. 2012;3:412.
Matsuno K, Yamada H, Iwata K, Jin D, Katsuyama M, Matsuki M, et al. Nox1 is involved in angiotensin II-mediated hypertension: a study in Nox1-deficient mice. Circulation. 2005;112:2677–85.
Yao H, Ferdaus MZ, Zahid HM, Ohara H, Nakahara T, Nabika T. Focal ischemic injury with complex middle cerebral artery in stroke-prone spontaneously hypertensive rats with loss-of-function in NADPH oxidases. PLoS ONE. 2015;10:e0138551.
Komers R, Xu B, Schneider J, Oyama TT. Effects of xanthine oxidase inhibition with febuxostat on the development of nephropathy in experimental type 2 diabetes. Br J Pharmacol. 2016; 2573–88.
Ishikawa N, Harada Y, Maruyama R, Masuda J, Nabika T. Genetic effects of blood pressure quantitative trait loci on hypertension-related organ damage: evaluation using multiple congenic strains. Hypertens Res. 2008;31:1773–9.
Nakamura T, Yamamoto E, Kataoka K, Yamashita T, Tokutomi Y, Dong YF, et al. Pioglitazone exerts protective effects against stroke in stroke-prone spontaneously hypertensive rats, independently of blood pressure. Stroke. 2007;38:3016–22.
Gandolgor TA, Ohara H, Cui ZH, Hirashima T, Ogawa T, Saar K, et al. Two genomic regions of chromosomes 1 and 18 explain most of the stroke susceptibility under salt loading in stroke-prone spontaneously hypertensive Rat/Izm. Hypertension. 2013;62:55–61.
Milatovic D, Montine TJ, Aschner M. Measurement of isoprostanes as markers of oxidative stress. Method Mol Biol. 2011;758:195–204.
Churchill PC, Churchill MC, Griffin KA, Picken M, Webb RC, Kurtz TW, et al. Increased genetic susceptibility to renal damage in the stroke-prone spontaneously hypertensive rat. Kidney Int. 2002;61:1794–1800.
El-Aal SAA, El-Fattah MAA, El-Abhar HS. CoQ10 augments rosuvastatin neuroprotective effect in a model of global ischemia via inhibition of NF-κB/JNK3/Bax and activation of Akt/FOXO3A/Bim cues. Front Pharmacol. 2017;8:735.
Wang Y, Chu C, Wang KK, Hu JW, Yan Y, Lv YB, et al. Effect of salt intake on plasma and urinary uric acid levels in Chinese adults: an interventional trial. Sci Rep. 2018;8:1434.
Kim-Mitsuyama S, Yamamoto E, Tanaka T, Zhan Y, Izumi Y, Izumiya Y, et al. Critical role of angiotensin II in excess salt-induced brain oxidative stress of stroke-prone spontaneously hypertensive rats. Stroke. 2005;36:1083–8.
Pires PW, Deutsch C, McClain JL, Rogers CT, Dorrance AM. Tempol, a superoxide dismutase mimetic, prevents cerebral vessel remodeling in hypertensive rats. Microvasc Res. 2010;80:445–52.
Horecky J, Gvozdjakova A, Kucharska J, E. Obrenovich M, H. Palacios H, Li Y, et al. Effects of coenzyme Q and creatine supplementation on brain energy metabolism in rats exposed to chronic cerebral hypoperfusion. Curr Alzheimer Res. 2011;8:868–75.
Abd-El-Fattah AA, El-Sawalhi MM, Rashed ER, El-Ghazaly MA. Possible role of vitamin E, coenzyme Q10 and rutin in protection against cerebral ischemia/reperfusion injury in irradiated rats. Int J Radiat Biol. 2010;86:1070–8.
Suzuki H, DeLano FA, Parks DA, Jamshidi N, Granger DN, Ishii H, et al. Xanthine oxidase activity associated with arterial blood pressure in spontaneously hypertensive rats. Proc Natl Acad Sci USA. 1998;95:4754–9.
Taheraghdam AA, Sharifipour E, Pashapour A, Namdar S, Hatami A, Houshmandzad S, et al. Allopurinol as a preventive contrivance after acute ischemic stroke in patients with a high level of serum uric acid: a randomized, controlled trial. Med Princ Pract. 2014;23:134–9.
Higgins P, Ferguson LD, Walters MR. Xanthine oxidase inhibition for the treatment of stroke disease: a novel therapeutic approach. Expert Rev Cardiovasc Ther. 2011;9:399–401.
Dawson J, Quinn T, Harrow C, Lees KR, Weir CJ, Cleland SJ, et al. Allopurinol and nitric oxide activity in the cerebral circulation of those with diabetes. Diabetes Care. 2009;32:135–7.
Waring WS, Webb DJ, Maxwell SR. Uric acid as a risk factor for cardiovascular disease. QJM. 2000;93:707–13.
Ma Y, Shen B, Zhang X, Lu Y, Chen W, Ma J, et al. Heritable multiplex genetic engineering in rats using CRISPR/Cas9. PLoS ONE. 2014;9:e89413.
The authors thank Satoko Mishima, Masamichi Koike and Masaki Misumi for their skillful assistance in the histological evaluation.
This work was partly supported by JSPS KAKENHI 26293086 (to T.N.) and 17K08787 (to H.O.).
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The authors declare that they have no conflict of interest.
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Ngarashi, D., Fujikawa, K., Ferdaus, M.Z. et al. Dual inhibition of NADPH oxidases and xanthine oxidase potently prevents salt-induced stroke in stroke-prone spontaneously hypertensive rats. Hypertens Res 42, 981–989 (2019). https://doi.org/10.1038/s41440-019-0246-2
- oxidative stress
- NADPH oxidase
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