Research Article| Volume 28, ISSUE 7, P1993-2002, July 2019

Clinical and Pathological Benefit of Twendee X in Alzheimer's Disease Transgenic Mice with Chronic Cerebral Hypoperfusion



      Multiple pathogeneses are involved in Alzheimer's disease (AD), such as amyloid-β accumulation, neuroinflammation, and oxidative stress. The pathological impact of chronic cerebral hypoperfusion on Alzheimer's disease is still poorly understood.


      APP23 mice were implanted to bilateral common carotid arteries stenosis with ameroid constrictors for slowly progressive chronic cerebral hypoperfusion (CCH). The effects of the administration of Twendee X (TwX) were evaluated by behavioral analysis, immunohistochemical analysis, and immunofluorescent histochemistry.


      In the present study, chronic cerebral hypoperfusion, which is commonly found in aged Alzheimer's disease, significantly exacerbated motor dysfunction of APP23 mice from 5 months and cognitive deficit from 8 months of age, as well as neuronal loss, extracellular amyloid-β plaque and intracellular oligomer formations, and amyloid angiopathy at 12 months. Severe upregulations of oxidative markers and inflammatory markers were found in the cerebral cortex, hippocampus, and thalamus at 12 months. Twendee X treatment (20 mg/kg/d, from 4.5 to 12 months) substantially rescued the cognitive deficit and reduced the above amyloid-β pathology and neuronal loss, alleviated neuroinflammation and oxidative stress.


      The present findings suggested a potential therapeutic benefit of Twendee X for Alzheimer's disease with chronic cerebral hypoperfusion.

      Key Words


      ACs (ameriod constrictors), AD (Alzheimer's disease), (amyloid-β), BCCAs (bilateral common carotid arteries), BSA (bovine serum albumin), CBF (cerebral blood flow), CCH (chronic cerebral hypoperfusion), CTX (cortex), DG (dentate gyrus), HI (hippocampus), M (months), PBS (phosphate-buffered saline), PFA (paraformaldehyde), Sub (subiculum), TH (thalamus), TwX (Twendee X), WT (wild type)
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        • Hishikawa N.
        • Fukui Y.
        • Sato K.
        • et al.
        Characteristic features of cognitive, affective and daily living functions of late-elderly dementia.
        Geriatr Gerontol Int. 2016; 16: 458-465
        • Selkoe D.J.
        Alzheimer's disease: genes, proteins, and therapy.
        Physiol Rev. 2001; 81: 741-766
        • Wang J.
        • Gu B.J.
        • Masters C.L.
        • et al.
        A systemic view of Alzheimer disease–insights from amyloid-β metabolism beyond the brain.
        Nat Rev Neurol. 2017; 13: 703
        • Perry G.
        • Nunomura A.
        • Hirai K.
        • et al.
        Oxidative damage in Alzheimer's disease: the metabolic dimension.
        Int J Dev Neurosci. 2000; 18: 417-421
        • Verdile G.
        • Keane K.N.
        • Cruzat V.F.
        • et al.
        Inflammation and oxidative stress: the molecular connectivity between insulin resistance, obesity, and Alzheimer's disease.
        Mediators Inflamm. 2015; 2015105828
        • Duncombe J.
        • Kitamura A.
        • Hase Y.
        • et al.
        Chronic cerebral hypoperfusion: a key mechanism leading to vascular cognitive impairment and dementia. Closing the translational gap between rodent models and human vascular cognitive impairment and dementia.
        Clin Sci. 2017; 131: 2451-2468
        • Shang J.
        • Yamashita T.
        • Zhai Y.
        • et al.
        Strong impact of chronic cerebral hypoperfusion on neurovascular unit, cerebrovascular remodeling, and neurovascular trophic coupling in Alzheimer's disease model mouse.
        J Alzheimers Dis. 2016; 52: 113-126
        • Zhai Y.
        • Yamashita T.
        • Nakano Y.
        • et al.
        Chronic cerebral hypoperfusion accelerates Alzheimer's disease pathology with cerebrovascular remodeling in a novel mouse model.
        J Alzheimers Dis. 2016; 53: 893-905
        • Kusaki M.
        • Ohta Y.
        • Inufusa H.
        • et al.
        Neuroprotective effects of a novel antioxidant mixture Twendee X in mouse stroke model.
        J Stroke Cerebrovasc Dis. 2017; 26: 1191-1196
        • Sturchler-Pierrat C.
        • Abramowski D.
        • Duke M.
        • et al.
        Two amyloid precursor protein transgenic mouse models with Alzheimer disease-like pathology.
        Proc Natl Acad Sci U S A. 1997; 94: 13287-13292
        • Ohta Y.
        • Nagai M.
        • Nagata T.
        • et al.
        Intrathecal injection of epidermal growth factor and fibroblast growth factor 2 promotes proliferation of neural precursor cells in the spinal cords of mice with mutant human SOD1 gene.
        J Neurosci Res. 2006; 84: 980-992
        • Kurata T.
        • Miyazaki K.
        • Kozuki M.
        • et al.
        Atorvastatin and pitavastatin improve cognitive function and reduce senile plaque and phosphorylated tau in aged APP mice.
        Brain Res. 2011; 1371: 161-170
        • Zhu X.
        • Tian J.
        • Sun S.
        • et al.
        (-)-SCR1693 protects against memory impairment and hippocampal damage in a chronic cerebral hypoperfusion rat model.
        Sci Rep. 2016; 6: 28908
        • Florian C.
        • Roullet P.
        Hippocampal CA3-region is crucial for acquisition and memory consolidation in Morris water maze task in mice.
        Behav Brain Res. 2004; 154: 365-374
        • Heneka M.T.
        • Kummer M.P.
        • Stutz A.
        • et al.
        NLRP3 is activated in Alzheimer's disease and contributes to pathology in APP/PS1 mice.
        Nature. 2013; 493: 674-678
        • Tobinick E.
        • Gross H.
        • Weinberger A.
        • et al.
        TNF-alpha modulation for treatment of Alzheimer's disease: a 6-month pilot study.
        MedGenMed. 2006; 8: 25
        • Perry G.
        • Cash A.D.
        • et al.
        Alzheimer disease and oxidative stress.
        J Biomed Biotechnol. 2002; 2: 120-123
        • Zhu X.
        • Lee H.G.
        • Casadesus G.
        • et al.
        Oxidative imbalance in Alzheimer's disease.
        Mol Neurobiol. 2005; 31: 205-217
        • Sultana R.
        • Butterfield D.A.
        Role of oxidative stress in the progression of Alzheimer's disease.
        J Alzheimers Dis. 2010; 19: 341-353
        • Hussain T.
        • Tan B.
        • Yin Y.
        • et al.
        Oxidative stress and inflammation: what polyphenols can do for us?.
        Oxid Med Cell Longev. 2016; 2016 (Epub 2016 Sep 22)7432797
        • Fischer R.
        • Maier O.
        Interrelation of oxidative stress and inflammation in neurodegenerative disease: role of TNF.
        Oxid Med Cell Longev. 2015; 2015 (Epub 2015 Mar 5)610813
        • Tamagno E.
        • Parola M.
        • Bardini P.
        • et al.
        Beta-site APP cleaving enzyme up-regulation induced by 4-hydroxynonenal is mediated by stress-activated protein kinases pathways.
        J Neurochem. 2005; 92: 628-636
        • Harrison F.E.
        • Allard J.
        • Bixler R.
        • et al.
        Antioxidants and cognitive training interact to affect oxidative stress and memory in APP/PSEN1 mice.
        Nutr Neurosci. 2009; 12: 203-218
        • Dumont M.
        • Kipiani K.
        • Yu F.
        • et al.
        Coenzyme Q10 decreases amyloid pathology and improves behavior in a transgenic mouse model of Alzheimer's disease.
        J Alzheimers Dis. 2011; 27: 211-223
        • Heo J.H.
        • Hyon-Lee
        • Lee K.M.
        The possible role of antioxidant vitamin C in Alzheimer's disease treatment and prevention.
        Am J Alzheimers Dis Other Demen. 2013; 28: 120-125
      1. Inufusa H. Composition for protection against cell-damaging effects, US Patent: 9089548 B2. Issued date July 28th, 2015.