Neuroprotective Effects of a Novel Antioxidant Mixture Twendee X in Mouse Stroke Model

https://doi.org/10.1016/j.jstrokecerebrovasdis.2017.01.003Get rights and content

Background

Oxidative stress and inflammation are important aggravating factors in acute ischemic stroke.

Methods

In the present study, the neuroprotective effects of a novel antioxidant mixture Twendee X containing multiple antioxidative ingredients, such as coenzyme Q10, ascorbic acid, and cystine, were evaluated. After the pretreatment of a vehicle or Twendee X (20 mg/kg/d) for 14 days, mice were subjected to transient middle cerebral artery occlusion for 60 minutes and further treated with vehicle or Twendee X for 1 or 5 days.

Results

Twendee X administration reduced the infarct size, and reduced oxidative stress markers such as 8-hydroxy-2′-deoxyguanosine, 4-hydroxy-2-nonenal, and Nε-(carboxymethyl) lysine (one of advanced glycation end products), as well as inflammatory markers such as ionized calcium binding adapter molecule-1, tumor necrosis factor-α, and monocyte chemotactic protein-1.

Conclusions

In the present study, the neuroprotective effects of Twendee X were shown on transient middle cerebral artery occlusion mice via antioxidative and anti-inflammatory pathways, providing a potential of Twendee X as one preventive and therapeutic treatment.

Introduction

Ischemic stroke is a leading cause of mortality and neurologic impairments worldwide.1 Therapeutic strategies against stroke remain limited, and further novel therapies are required in daily clinical practice.2 Oxidative stress and inflammation are important aggravating factors in acute ischemic stroke.3, 4 Reactive oxygen species (ROS) is gradually generated during cerebral ischemia, then excessively increased after reperfusion notably in the peri-ischemic area with oxidation of cellular DNA, lipids, and proteins,5, 6, 7, 8 and then often focused as a medical treatable target.9

We originally showed the strong neuroprotective effects of a free radical scavenger edaravone.8, 10, 11 Furthermore, we reported that dietary supplements such as ginkgo extract,12 platinum nanoparticle species,13 and antioxidativenutrient-rich enteral diet14 showed neuroprotective effects on the ischemic brains of mice. Dietary supplements did not show the strong effects on diseases as medicinal chemicals. However, they are safe and valuable for the prevention and treatment of various diseases.15

Twendee X (TwX) is an anti-aging supplement containing multiple antioxidants and a patented composition.16 TwX has strong antioxidant effects, and increases superoxide dismutase and cell protection effects.17 In the present study, we validated whether TwX could be helpful in ameliorating mouse brain damages and oxidative stress following experimental transient middle cerebral artery occlusion (tMCAO).

Section snippets

Animals and Focal Cerebral Ischemia

All experimental procedures were approved by the Animal Committee of the Okayama University Graduate School of Medicine (OKU-2015573). Adult male C57BL/6JJcl mice (23-27 g, 8 weeks old) were obtained from CLEA Japan (Tokyo, Japan). The mice were maintained in a temperature-regulated room (23-25°C) on a 12-hour light–dark cycle and allowed free access to food and water. From 9 weeks of age, the mice received vehicle (physiological saline, ip, n = 15) or TwX (20 mg/kg per day, ip, n = 16) for 14

Serum Oxidative Stress and the Antioxidative Activities

As compared with the vehicle (V-1d, 165 ± 21.3 CARR U), TwX did not reduce oxidative stress (d-ROMs) 1 day after tMCAO (TwX-1d, 164 ± 10.3 CARR U), but showed a slight reduction 5 days after (V-5d = 167.5 ± 27.8 versus TwX-5d = 149.5 ± 14.4 CARR U, not significant) (Fig 2). On the other hand, OXY-Adsorbent showed a reduction tendency in TwX both 1 day after tMCAO (V-1d, 319.0 ± 52.4 µmol HClO/mL; TwX-1d, 294.0 ± 33.4 µmol HClO/mL) and 5 days after (V-5d, 337.5 ± 51.3 µmol HClO/mL; TwX-5d,

Discussion

The present study demonstrated that a novel antioxidant mixture TwX showed neuroprotective effects on mice cerebral ischemia by reducing the infarct size (Fig 3), and reducing both oxidative stress markers (Fig 4) and inflammatory markers (Fig 5). ROS is incrementally generated during cerebral ischemia, then explosively increased after reperfusion particularly in the penumbra with oxidation of cellular DNA, lipids, and proteins.5, 6, 7, 8 Such oxidative stress is detrimental in the cerebral

Acknowledgments

We are grateful to Dr. Maruyama T of the Department of Preventive Dentistry at Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, especially for their technical assistance in d-ROMs tests and OXY-Adsorbent tests.

References (31)

  • T. Morimoto et al.

    Simultaneous measurement of salicylate hydroxylation and glutamate release in the penumbral cortex following transient middle cerebral artery occlusion in rats

    J Cereb Blood Flow Metab

    (1996)
  • K. Abe

    Therapeutic potential of neurotrophic factors and neural stem cells against ischemic brain injury

    J Cereb Blood Flow Metab

    (2000)
  • ZhangW. et al.

    Extension of ischemic therapeutic time window by a free radical scavenger, Edaravone, reperfused with tPA in rat brain

    Neurol Res

    (2004)
  • O. Peters et al.

    Increased formation of reactive oxygen species after permanent and reversible middle cerebral artery occlusion in the rat

    J Cereb Blood Flow Metab

    (1998)
  • K. Abe et al.

    Strong attenuation of ischemic and postischemic brain edema in rats by a novel free radical scavenger

    Stroke

    (1988)
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    Grant support: This work was partly supported by Grants-in-Aid for Scientific Research (B, 25293202 and C, 15K09316), Challenging Research (15K15527), and Young Research (15K21181), and by Grants-in-Aid from the Research Committees (Mizusawa H, Nakashima K, Nishizawa M, Sasaki H, and Aoki M) from the Ministry of Health, Labour and Welfare, Japan.

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