Table 2.
Preclinical studies of Nrf2 in transient cerebral ischemia models of mice and rats (MCAO, 40 min-1.5 h).
| Model | Species | Genetic Background; Sex; Age or Weight | Treatment | Dosage/Administration route | Brain lesion/Edema | Neurobehavioral deficits | Nrf2 Mechanism (in vivo) | References |
|---|---|---|---|---|---|---|---|---|
| MCAO (40 min-1.5 h) | ||||||||
| Findings supporting the role of Nrf2 pathway (with Nrf2−/− mice) | ||||||||
| MCAO (40 min) | Mouse | C57BL/6, Nrf2−/−, and Cx3cr1GFP/+; M; 23–30 g | 3H-1,2-Dithiole-3-thione (D3T) | • Post; 50 mg/kg; i.p.; at 3 h after MCAO • Once |
• Infarct volume (48 h) • Brain edema (48 h) • Above changes are decreased in Nrf2−/− mice |
• Neurological deficits (48 h) ↓ • Above changes are absent in Nrf2−/− mice |
• Nrf2 (total ↑) and HO1 proteins (in vitro) • Above changes are absent in Nrf2−/− mice • Nrf2 (IHC) |
Kuo et al., 2017 |
| MCAO (1 h) | Mouse | C57BL/6J WT, Nrf2−/−; M; 7–11wks; | Resveratrol | • Pre: 10 mg/kg; i.p.; 48 h before MCAO • Once |
• Infarct volume (24 h) ↓ • Above change is absent in Nrf2−/− mice |
• NA | • NQO1, SOD2 proteins • Above changes are decreased in Nrf2−/− mice |
Narayanan et al., 2015 |
| MCAO (1 h) | Mouse | C57BL/6J: Nrf2−/− and WT; M; 8–10wks | Lentiviral transfection (for SIRT6 overexpression) | • Pre; 2.5 μl (109 infectious units/ml); i.c.v.; 2 wks before MCAO • Once |
• Infarct volume (24 h) ↓ • Above change is absent in Nrf2−/− mice |
• Neurological deficits (24 h) ↓ • Above change is absent in Nrf2−/− mice |
• Nrf2 (total ↑) and HO1 proteins | Zhang et al., 2017 |
| MCAO (1 h) | Mouse | C57BL/6: WT and Nrf2−/−; 20–25 g, 8 to 10 wks | Dimethyl fumarate (DMF), Monomethyl fumarate (MMF) | • Post: 30, 45 mg/kg (better); i.p.; 15 min before reperfusion twice a day • For 7 d |
• Infarct volume (3,7 d) ↓ • Brain edema (3 d) ↓ • Above changes are absent in Nrf2−/− mice |
• Neurological deficits (3,7 d) ↓ • Above change is absent in Nrf2−/− mice |
• Nrf2 (total ↑) and HO1 proteins | Yao et al., 2016 |
| MCAO (1 h) | Mouse | WT and Nrf2−/−; M; 8–10 wks | Tanshinone IIA (TSA) | • Post; 25 mg/kg; i.p.; 10 min after MCAO • Once |
• Infarct volume (72 h) • Above change is reduced in Nrf2−/− mice |
• Neurological deficits (72 h) ↓ • Above change is absent in Nrf2−/− mice |
• Nrf2 (nuclear ↑) protein • Nrf2 mRNA |
Cai et al., 2017 |
| MCAO (1 h) | Mouse | ICR background Nrf2−/− and WT; 25–28 g | Ursolic acid (UA) | • 130 mg/kg; i.p.; immediately after MCAO; once | • Infarct volume (24 h) ↓ • Above change is absent in Nrf2−/− mice |
• Neurological deficits (24 h) • Above change is reduced in Nrf2−/− mice |
• Nrf2 (nuclear ↑, cytoplasmic ↓) and HO1 proteins • Nrf2 and HO1 mRNA |
Li et al., 2013 |
| MCAO (1.5 h) | Mouse | HO1−/−, Nrf2−/− and WT; M; 7–8 wks; 20–25 g | Epicatechin (EC) | • Pre: 2.5 (no effect), 5, 15, 30 mg/kg (best); oral; 90 min before; once • Post: 30 mg/kg EC was administered at 3.5 h (better) or 6 h (no effect) after MCAO |
• Infarct volume (pre; 24 h) ↓ • Above change is absent in Nrf2−/− mice • Infarct volume (post; 72 h) ↓ |
• Neurological deficits (pre; 24 h) • Above change is reduced in Nrf2−/− mice • Neurological deficits (post; 72 h) |
• Nrf2 (nuclear ↑, cytoplasmic) and HO1 proteins | Shah et al., 2010 |
| MCAO (1.5 h) | Mouse | CD1 background Nrf2−/− and WT; F; 20–25 g | Tert-butylhydroquinone (t-BHQ) | • NA | • Infarct volume (between Nrf2−/− and WT, 24 h) • Above change is reduced in Nrf2−/− mice |
• Neurological deficits (between Nrf2−/− and WT; 24 h) • Above change is reduced in Nrf2−/− mice |
• NA | Shah et al., 2007 |
| Findings supporting the role of Nrf2 pathway (without Nrf2−/− mice, indicated by Nrf2 protein nuclear translocation) | ||||||||
| MCAO (1 h) | Mouse | ICR; M; 24–27 g | Isorhamnetin (Iso) | • Post; 5 mg/kg; i.p.; immediately at the onset of reperfusion; daily; • Twice |
• Infarct volume (48 h) ↓ • Brain edema (48 h) |
• Neurological deficits (48 h) ↓ • Rotarod (48 h) |
• Nrf2 (nuclear ↑, cytoplasmic) and HO1 proteins | Zhao et al., 2016 |
| MCAO (1.5 h) | Mouse | C57BL/6; M; 25–30 g; 10–12 wks | Epigallocatechin-3-gallate (EGCG) | • Post; 50 mg/kg; i.p.; immediately after; daily • For 7 d |
• Infarct volume (7 d) ↓ | • Neurological deficits (3, 7 and 14 d) ↓ | • Nrf2 (nuclear ↑) protein • Nrf2 (IHC) |
Bai et al., 2017 |
| MCAO (1.5 h) | Rat | SD; M; 280–300 g | Diterpene ginkgolides meglumine injection (DGMI) | • Post; 1, 3 and 10 mg/kg, iv, at the onset of reperfusion and 12 h after reperfusion | • Infarct volume (3 and 10 mg/kg, 24 h) ↓ | • Neurological deficits (1, 3 and 10 mg/kg dose-dependent, 24 h) ↓ | • Nrf2 (nuclear ↑) and HO1 | Zhang W. et al., 2018 |
| MCAO (1 h) | Rat | SD; M; 3 mo | 5-methoxyindole-2-carboxylic acid (MICA) | • Pre; diet supplemented with 0.33% MICA (200 mg/kg/d) for 4 wks before MCAO; i.p. injection (200 mg/kg body weight) once per day • For seven days until 24 h before MCAO |
Infarct volume (24 h) ↓ | • NA | • Nrf2 (nuclear ↑) and NQO1 proteins | Wu et al., 2017b |
| Other findings involving the role of Nrf2 pathway | ||||||||
| MCAO (1 h) | Mouse | C57BL/6J background WT and SHPS-1 mutant (MT); M; 10–12 wks | Src homology 2 domain–containing protein tyrosine phosphatase substrate−1 (SHPS-1) | • NA | • Infarct volume (24 h) ↓ | • Neurological deficits (72 h) ↓ | • Nrf2 (total ↑; at 98 KDa) and HO1 proteins | Wang B. et al., 2011 |
| MCAO (1 h) | Mouse | C57BL/6 J; M; 22–25 g | MiR-93 antagomir | • Pre; 7 μl (at 100 μm); i.c.v.; 10 min before MCAO • Once |
• Infarct volume (24 h) ↓ | • Neurological deficits (24 h) ↓ | • Nrf2 (total ↑) and HO1 proteins | Wang et al., 2016 |
| MCAO (1 h) | Mouse | ICR; M; 6 wks old, 23–25 g | Tocovid | • 200 mg/kg/d orally once a day for 1 mo before MCAO | • Infarct volume (1, 3 d) ↓ | • Bederson score (pre,1,3,7 d), • Rotarod (pre,1,3,7 d) • Corner (pre,1,3,7 d) |
• Nrf2 (total ↑) protein • Nrf2 (IHC) |
Shang et al., 2018 |
| MCAO (1 h) | Mouse | C57BL/6 J | Gastrodin (GAS) | • Post: 10, 50, 100 mg/kg; i.p.; onset of cerebral reperfusion; • Once daily for 7 d |
• Infarct volume (medium or high-dose, 24 h and 7 d) ↓ | • Neurobehavioral scores (1, 7 d) ↓ | • Nrf2 (total ↑; at 68 KDa) and HO1 proteins | Peng et al., 2015 |
| MCAO (1 h) | Mouse | C57BL/6; F; 12–15 wks | Estradiol (EST) | • 0.05 mg; pellets; subcutaneous implantation; before; for 21 d | • Brain edema (24 h) ↓ | • Neurological deficits (mNSS, 24 h) ↓ | • Nrf2 (total ↑) and NQO1 proteins | Li et al., 2017 |
| MCAO (1 h) | Rat | SD; M; 60–80 d old, 260–300 g | Tert-butylhydroquinone (tBHQ) | • Pre; 16.7 mg/kg; i.p. injection at intervals of 8 h before MCAO • Three times |
• Infarct volume (24 h) | • Neurological deficits (24 h) ↓ | • Nrf2 (total ↑) protein | Hou et al., 2018 |
| MCAO (70 min) | Rat | SD; M; 250–330 g | Sulforaphane | • 5 mg/kg; i.p.; 1 h before; once | • Infarct volume (24 h, 72 h) | • Neurological deficits (24 h) ↓ | • Nrf2 (total content ↑) and NQO1 proteins • HO1 (IHC) |
Alfieri et al., 2013 |
| MCAO (1 h) | Rat | SD; M; 3 mo | 5-methoxyindole-2-carboxylic acid (MICA) | • Post; 100 mg/kg;i.p.; at the onset of reperfusion • Once |
• Infarct volume (24 h) ↓ | • NA | • Nrf2 (total ↑) and NQO1 proteins | Wu et al., 2018 |
| MCAO (1.5 h) | Rat | SD; F; 250–300 g | Genistein | • Pre; 10 mg/kg, i.p., once daily • For 2 wks |
• Infarct volume (72 h) ↓ | • Neurological deficits (72 h) ↓ | • Nrf2 (total ↑) and NQO1 proteins | Miao et al., 2018 |
| MCAO (1 h) | Rat | SD; M; 60–80 d old, 240–300 g | Glycogen synthase kinase 3β (GSK-3β) | • Pre: 7 μl (2 μg/μl); i.c.v.; 48 h before MCAO • Once |
• NA | • NA | • Nrf2 (total) and NQO1, HO1 proteins • NQO1, HO1 mRNA |
Chen X. et al., 2016 |
| MCAO (1 h) | Rat | SD; M; 250–280 g | siRNA targeting sulfiredoxin1 (Srxn1) | • Pre; i.c.v.; 24 h before MCAO • Once |
Infarct volume (24 h) | • Neurological deficits (24 h) ↑ | • Nrf2 (total ↓) and NQO1 proteins | Wu et al., 2017a |
| MCAO (1 h) | Rat | SD; M; 270–310 g | Thioredoxin-1 siRNA | • Pre: 10 μl (2 μg/μl); i.c.v.; 24 h before MCAO • Once |
• Infarct volume (24 h) • Brain edema (24 h) |
• Neurological deficits (24 h) ↓ | • Nrf2 (total ↓) protein | Li et al., 2015 |
| MCAO (1 h) | Rat | SD; M; 280–310 g | Sevoflurane | • Post: 2.6% for 1 h; inhalation; immediately at onset of reperfusion • Once |
• Infarct volume (72 h) | • Neurological deficits (12, 24, 48, and 72 h) n | • Nrf2 (total ↓) and NQO1 • Nrf2-DNA binding activities |
Li et al., 2014 |
| MCAO (70 min) | Rat | SD; M; 250–300 g | Nrf2 inducer D, L-sulforaphane | • 5 mg/kg; i.p.; 1 h before MCAO • Once |
• NA | • NA | • Nrf2 (IHC) | Srivastava et al., 2013 |
| MCAO 1 h | Rat | SD; M; aged 8–9 wks; 300–350 g | • MicroRNA (miR-142-5p) | • NA | • NA | • NA | • Nrf2 mRNA | Wang et al., 2017 |
| MCAO (1 h) | Rat | Wistar; M; 250–280 g | Danhong | • 0.9, 1.8 ml/kg; i.p.; 30 min before ischemia, with reperfusion and 24, 48, 72 h after ischemia | • Infarct volume (72 h) ↓ • Brain edema (72 h) ↓ |
• Neurological deficits (72 h) | • Nrf2 and HO1, NQO1 mRNA | Guo et al., 2014 |
| MCAO (1.5 h) | Rat | SD; M; 180–220 g | Lactulose | • Pre; 0.25 g/kg; gavage; at start of ischemia • Once |
• Infarct volume (24 h) ↓ | • Neurological deficits (24 h) ↓ • Morris water maze ↓ |
• Nrf2 mRNA and activity • SOD activity |
Zhai et al., 2013 |
| MCAO (1.5 h) | Rat | SD; M; 260–290 g | β-caryophyllene (BCP) | • Pre; 34, 102, 306 mg/kg (best); gavage; once a day • For 7 d |
• Infarct volume (24 h) ↓ | Neurological deficits (24 h) ↓ | • Nrf2 (total ↑) and HO1 mRNA | Lou et al., 2016 |
| MCAO (1.5 h) | Rat | SD; M; 260–280 g | Neural stem cells (NSCs) | • Post; four 1.0 μl deposits of single-cell suspension in Dulbecco's PBS (105 cells per ul); along the anterior-posterior axis into the cortex; 6 h after stroke | • Infarct volume (cortex, 28 d) ↓ | • Rotarod (1–28 d) • Beam-balance (28 d) ↓ |
Nrf2 mRNA | Sakata et al., 2012 |
| MCAO (1.5 h) | Rat | Wistar; M; 250–280 g | Xueshuantong injection (Lyophilized, XST) | • Post; 25, 50,100 mg/kg; i.p.; 1 h after reperfusion, once a day • For 3 or 7 d |
• NA | • Modified neurological severity (Mnss, 1, 3, and 7 d) ↓ | • Nrf2 and HO1, NQO1 mRNA | Guo et al., 2018 |
| MCAO (1 h) | Rat | SD; F; 300–350 g | p-hydro-xybenzyl alcohol (HBA) | • Pre; 25 mg/kg BW; i.m. with sesame oil; 3 d before • Once |
• Infarct volume (cortex and striatum, 24 h) ↓ | • Modified neurological severity score (mNSS) at 1, 7, 14, 21, and 28 d • Functional deficits from 7 d ↓ |
• Nrf2 DNA (PCR) | Kam et al., 2011 |
| MCAO (1 h) | Rat | SD; M; 230–270 g | Curcumin | • Post; 300 mg/kg; i.p.; 1 h after MCAO • Once |
• Infarct volume (24 h) ↓ | • NA | • Nrf2-DNA binding activity | Wu J. et al., 2013 |
| MCAO (45 min) | Mouse | OKD48 transgenic mice; M/F; 23–28 g | NA | • NA | • Infarct volume (12 h, 1, 3, 7 d) | • NA | • Nrf2 (IF) | Nakano et al., 2017 |
| MCAO (1 h) | Mouse | ICR; M; 34–38 g; 8 wks | NA | • NA | • NA | • NA | • Nrf2 (IHC) | Tanaka et al., 2011 |
| MCAO (1 h) | Rat | Hannover-Wistar; M; 250–350 g | Recombinant human erythropoietin (rhEpo) | • Post; 5000 IU/kg; i.p. immediately or 3 h after MCAO • Once |
• Infarct volume (3, 24 h) | • NA | • Nrf2 (IHC) | Mrsic-Pelcic et al., 2017 |
| MCAO (1 h) | Rat | Wistar; M; 10 wks; 250–300 g | Curcumin | • Post; 300 mg/kg; i.p.;Post; 300 mg/kg; i.p.; 30 min after MCAO • Once |
• Infarct volume (d1) ↓ • Brain edema (d1) ↓ |
• Neurological deficits (d1) ↓ | • Nrf2 (IHC) | Li et al., 2016 |
| MCAO (1 h) | Mouse | Transgenic fatty acid metabolism-1 (fat-1) gene mice; C57BL/6; M | Omega-3 fatty acids (n-3 PUFAs) by fish oil (FO) diet | • Pre: 5% (w/w) was added to the regular diet, which increased the n-3 PUFA from 0.34 to 1.5%, and decreased the n-6:n-3 PUFA ratio from 5:1 to 1:1; oral; before; daily; for 6 wks | • Infarct volume (48 h) ↓ | • Neurological deficits (48 h) ↓ • Dietary supplementation in Corner, Rotarod (7 d) d supplementation in Corner, Rotarod |
HO1 protein | Zhang M. et al., 2014 |
| MCAO (1 h) | Mouse | C57BL/6; M; 8-−0 wks | Dimethyl fumarate (DMF) | • Pre: 15 mg/kg; i.p.; twice a day for 3 d before stroke | • Infarct volume (4 h, 24 h) • Brain edema (4 h, 24 h) ↓ |
• NA | • HO1, NQO1,GCLC and GCLM mRNA | Kunze et al., 2015 |
| MCAO (1.5 h) | Rat/ Mouse | Wistar (Osmotic pump studies) and SD, 250–350 g; | Tert-butylhydroquinone (tBHQ) | • Rat: 1 mM, i.c.v., osmotic mini-pump delivery (1 μl/h for 4 d), MCAO after 3 • d; i.p. in later experiments, 3.33 or 16.7 mg/kg, before; three times by 8 h intervals |
• Rat: Infarct volume (cortex, 24 h) ↓ | • Rat: tBHQ Neurological deficits (24 h to 1 mo) • Sensorimotor deficits (since 4 d) ↓ |
NA | Lou et al., 2016 |
MCAO, middle cerebral artery; pdMCAO, permanent distal middle cerebral artery occlusion; pMCAO, permanent (proximal end of) middle cerebral artery occlusion that is generated by the intraluminal suture MCAO; GCI, global cerebral ischemia; i.p., intraperitoneal; i.v., intravenous; i.c.v., intracerebroventricular; Pre, pretreatment; Post, posttreatment; the changes in brain lesion/edema and neurobehavioral deficits/Nrf2 protein expression level rf2 protein expression leveledema and neurobehavioral deficitseatmest, posttreatmeatme; IF, Immunofluorescence; KDa, kilodalton that indicates Nrf2 protein molecular weight by Western blot; the changes in brain lesion, edema, neurological deficits, and mRNA/protein expression level (↑ or ↓, increase or decrease at indicated time point; no label at indicated time point, no significant difference); h, hour; d, day; wk, week.