Table 5.
Anti-inflammatory treatments in ischemic stroke: evidence from preclinical studies.
| Study | Year | Treatment | Sample Size | Outcome |
|---|---|---|---|---|
| IL-1Ra | ||||
| Garcia et al. [256] | 1995 | 13 with pMCAO and treated with IL-1Ra; 13 with pMCAO and treated with CSE buffer or placebo group (n = 13); 2 sham-operated animals treated with IL-1Ra or CSE (n = 2) | 30 outbred male Wistar rats and fed Agway rat chow during the 4–6 quarantine days | IL-1Ra in rats with pMCAO significantly decreased the number of necrotic neurons both at 24 h and 7 days after the arterial occlusion (p < 0.0001). Neurological scores were also significantly improved with and a non-significant decrease in the number of PMN leukocytes in the ischemic hemisphere was observed. |
| Yamasaki et al. [247] | 1995 | 60 min of tMCAO followed by reperfusion; first IL-1β, then anti-IL1β was injected | 120 adult male Wistar rats | tMCAO induced an increase in brain water content, necrosis, and neutrophilic infiltration in the cortex perfused by the MCA and the DCP and VCP. rIL-1β into the left lateral ventricle immediately after reperfusion markedly enhanced ischemic brain edema formation and infarction size in MCA zone, DCP, and VCP in a dose-dependent manner (p < 0.01). Anti-IL-1β attenuated the post-ischemic increase of brain water content and decreased the infarction size (p < 0.01). The number of neutrophils infiltrating the ischemic area decreased with anti-IL-1β. |
| Relton et al. [236] | 1996 | MCAO or sham surgery. Animals were injected subcutaneously with either vehicle or rIL-1Ra at 0, 4, 8, 12, and 18 h after ischemia. In separate experiments, initial treatment was delayed until 30 min, 1 h, or 4 h after ischemia and treatments were repeated until 18 h | Male Sprague-Dawley rats | rIL-1Ra significantly inhibited infarct size by 46% at 24 h (p < 0.05), cerebral edema formation by 49% at 24 h (p < 0.05). Infarction inhibition by rIL-1Ra was dependent on dose and time of administration. |
| Pradillo et al. [234] | 2012 | Lean and Cp rats received placebo or IL-1Ra (25 and 12.5 mg/kg) subcutaneously at reperfusion and 6 h later and allocated to different groups: lean + tMCAO + placebo; lean + tMCAO + IL-1Ra; Cp + tMCAO + placebo; and Cp + tMCAO + IL-1Ra. For the delayed administration study, animals were injected subcutaneously with placebo or IL-1Ra at 3 h of reperfusion and again 3 h later | Male, lean and Cp rats | IL-1Ra at reperfusion resulted in a 50% reduction of infarct volume as measured by MRI both in lean and Cp compared with placebo-treated animals (p < 0.05). IL-1Ra decreased the number of MMP-9-positive neutrophils when compared with placebo (p < 0.05). In both lean and Cp rats, IL-1Ra largely reduced the microglial activation compared with the placebo-treated groups (p < 0.05). In 16-month-old lean rats, delayed IL-1Ra significantly reduced the number of MMP-9-positive blood vessels and the number of MMP-9-positive neutrophils when compared with the placebo group (p < 0.05). |
| Statins | ||||
| Endres et al. [219] | 1998 | After tMCAO followed by reperfusion, mice were injected subcutaneously with 0.1 mL of activated simvastatin or lovastatin (0.2–20 mg/kg) or a corresponding volume of PBS once daily for 3 or 14 days | Not declared | In a concentration-dependent manner, simvastatin for 14 days reduced cerebral infarct size by 18, 27 and 46% (p < 0.05) and increased NOS activity (p < 0.05). Simvastatin 20 mg/kg increased basal hemispheric CBF by 31% (p < 0.05). Lovastatin 20 mg/kg daily for 14 days also decreased cerebral infarct size and neurological deficits, even if to a lesser extent than simvastatin. |
| Kawashima et al. [222] | 2003 | Two groups, one statin-treated (cerivastatin 2 mg/kg by gavage once daily) and another vehicle-treated | Stroke-prone spontaneously hypertensive rats (4 weeks of age) | The incidence of stroke and stroke size decreased (p < 0.01). High-dose statin treatment delayed early death and reduced the occurrence of stroke-associated symptoms (p < 0.01) and decreased stroke-associated infiltration of inflammatory cells (p < 0.05). Statin treatment increased eNOS protein levels and eNOS activity (p < 0.05). Superoxide production was reduced in statin-treated rats (p < 0.01). |
| Amin-Hanjani et al. [211] | 2001 | Two groups: mevastatin at a dose of 2 or 20 mg/kg daily and a corresponding concentration of vehicle for 7, 14, or 28 days before tMCAO | Wild-type male mice and eNOS-deficient male mice | Mevastatin increased levels of eNOS mRNA and protein, reduced infarct size, and improved neurological deficits in a dose- and time-dependent manner especially with 14- and 28-day high-dose treatment (26% and 37% infarct reduction, respectively, p < 0.05). Cholesterol levels were reduced only after 28 days of treatment (p < 0.05), but did not correlate with infarct reduction. Baseline absolute cerebral blood flow was 30% higher after 14-day high-dose treatment (p < 0.05). |
| Prinz et al. [235] | 2008 | After tMCAO followed by reperfusion, mice were treated with intravenously or intraperitoneally rosuvastatin given up to 6 h after MCAO (0.02–20 mg/kg) | Wild-type mice aged 6 to 8 weeks | Intravenous rosuvastatin significantly reduced lesion size up to 4 h after MCAO in doses as low as 0.2 mg/kg (p < 0.05). Intraperitoneal administration provided protection only on reperfusion at a dose of 20 mg/kg (p < 0.05). Lesion protection was evident 5 days after brain ischemia and was associated with functional improvements at 2.0 mg/kg dose (p < 0.05). Neuroprotection with intravenous rosuvastatin was achieved with peak plasma concentrations <0.5 ng/mL and was associated with increased levels of phosphorylated Akt kinase and eNOS in the vasculature (p < 0.05). |
| Asahi et al. [212] | 2005 | Heterologous blood clots were used to induce MCAO after long-term simvastatin (20 mg/kg), atorvastatin (20 mg/kg) or vehicle treatment subcutaneously | Male SV-129 mice and male C57Bl/6 mice | In wild-type mice, both simvastatin and atorvastatin reduced ischemic lesions and residual clot after 14 days (p < 0.05). In eNOS knockout mice, atorvastatin reduced the volume of ischemic tissue and improved neurologic outcomes after arterial occlusion (p < 0.05). Both statins did not have protective effects in t-PA knockout mice after embolic focal ischemia, but only in a filament model where focal ischemia was achieved via mechanical occlusion (p < 0.05). |
| Chen et al. [215] | 2003 | 24 h after MCAO, rats were fed atorvastatin (1, 3 or 8 mg/kg) daily for 7 days. Rats were also treated with simvastatin 1 mg/kg with the same protocol | 48 Adult male Wistar rats | Rats treated with 1 and 3 mg/kg atorvastatin and 1 mg/kg simvastatin improved functional recovery (p < 0.05). VEGF production within the ischemic boundary area at 14 days after stroke increased in the 1 mg/kg atorvastatin group (p < 0.05) as well as cyclic guanosine monophosphate, angiogenesis, neurogenesis, and synaptophysin levels (p < 0.05). |
| Sironi et al. [242] | 2003 | Two groups of rats were treated with vehicle alone or simvastatin for 3 days before MCAO, while other two groups underwent MCAO and were treated with vehicle or simvastatin at 3 and 25 h after the induction of the injury. The brain infarct size was evaluated using MRI | Male Sprague-Dawley rats | Treatment with simvastatin (20 mg/kg) after MCAO prevented the increase in brain infarct volume occurring at 24 h and induced a 46.6% reduction after 48 h (p < 0.01). The neuroprotective effects of simvastatin were paralleled by an increase in eNOS immunoreactivity, detectable in the brain of simvastatin-treated rats. |
| Reuter et al. [238] | 2015 | Cultured hBMECs pretreated with simvastatin and subjected to OGD | hBMECs | Simvastatin significantly blocked the expression of MMP-2 under OGD (p < 0.004). MMP-9 synthesis rate was low and unaffected by simvastatin treatment, while the gene expression and protein secretion of TIMP-1 and TIMP-2 were both strongly induced (p < 0.001). |
| Fingolimod (FTY720) | ||||
| Rolland et al. [239] | 2013 | Fingolimod was given intraperitoneally at a dose of 1 mg/kg as single dose 1 h after ICH induction or daily administration 1, 24, and 48 h after ICH induction | 103 male CD-1 mice and 28 male Sprague-Dawley rats | Fingolimod enhanced neurological functions and reduced brain edema at 24 and 72 h following experimental ICH in CD-1 mice (p < 0.05). Fewer lymphocytes were found in blood and brain samples of treated animals (p < 0.05). Fingolimod decreased ICAM-1, IFN-γ, IL-17 levels 72 h after ICH (p < 0.05). Treated Sprague-Dawley rats showed less spatial and motor learning deficits along with significantly reduced brain atrophy and neuronal cell loss within the basal ganglia (p < 0.05). |
| Campos et al. [214] | 2013 | 3 cohorts: pMCAO not treated with t-PA; tMCAO followed by early (30 min after thrombin) t-PA administration; and tMCAO followed by delayed (3 h after thrombin) t-PA administration. Each of these cohort received fingolimod at different time points | C57BL/6 male mice | Fingolimod reduced neurological deficits and infarct volume after in situ thromboembolic MCAO (p < 0.05). Combination of fingolimod and t-PA improved neurological outcomes of the thrombolytic therapy and the risk of hemorrhagic transformation associated with delayed administration of t-PA (p < 0.05). |
| Donepezil | ||||
| Wang et al. [244] | 2014 | 3 groups: the sham operation group (SO), the model group (MG) and the treatment group (TG). Pathological appearance of the hippocampal CA1 region and calpain I and CDK5/p25 expression were observed on the 4th, 6th and 8th week from I/R surgery | 250 3-month old male mice | At each postoperative time point, the normal neuron count of the hippocampal CA1 region in the treatment group increased significantly (p < 0.05), whereas calpain I and CDK5/p25 expression, SOD activity and MDA content were significantly lower than those in the model group (p < 0.05). |
| Min et al. [231] | 2012 | After transient global ischemia, donepezil (5 mg/kg once a day) was administered intragastrically for 21 days | Male Mongolian gerbils | Donepezil significantly inhibited delayed neuronal death in the hippocampal CA1 region (p < 0.01). Memory impairment was significantly improved by donepezil treatment (p < 0.05–0.01). Western blot analysis showed that donepezil treatment prevented reductions in p-CaMKII and p-CREB protein levels in the hippocampus (p < 0.01). |
| Yuan et al. [252] | 2011 | Cultured cells were exposed to both OGD and electrophysiological experiment | HEK293 cells from a human embryonic kidney cell line | Donepezil showed to attenuate OGD-induced apoptosis in Kv2.1/HEK293 cells and to inhibit Kv2.1 currents in a dose-dependent manner under normoxic condition (p < 0.01). Donepezil further inhibited Kv2.1 currents after OGD treatment (p < 0.05). |
| Akasofu et al. [210] | 2008 | Prolonged opening of sodium channels with veratridine led to depolarization-induced neuronal cell injury, which was prevented by 0.1 µM tetrodotoxin | Cortical cell cultures from fetal rats of the Wistar strain | Pre-treatment with donepezil (0.1–10 µM) for 1 day significantly decreased cell death and increased cell viability in a concentration-dependent manner (p < 0.05). At 0.1–10 µM, donepezil concentration-dependently decreased the veratridine-induced increase of calcium concentration, whilst at 10 µM it reduced the veratridine-induced increase of sodium concentration (p < 0.05 for both). |
| Lee et al. [225] | 2007 | After permanent ligation of bilateral common carotid arteries, rats were administered cilostazol (30 mg/kg/day orally) and donepezil (0.3 mg/kg/day intraperitoneally) | Rats | Concurrent treatment with cilostazol and donepezil prevented neuropathological alterations in the white matter by activation of phosphorylated CREB and Bcl-2, resulting in improvement of spatial learning memory (p < 0.05). |
| Citalopram | ||||
| Espinera et al. [220] | 2013 | After focal ischemic stroke, citalopram 10 mg/kg was injected intraperitoneally 24 h after stroke and then daily for 7, 14, 21, or 28 days | Adult male C57 mice | Citalopram had no significant effect on infarct formation or edema 3 days after stroke, but enhanced sensorimotor functional recovery after 14 days (p < 0.05). Citalopram improved neuroblast proliferation and migration (p < 0.01) as well as neurogenesis (p < 0.05) and peri-infarction vessel density (p < 0.05) in the post-ischemic brain. |
| Kronenberg et al. [223] | 2012 | Mice were subjected to 30-min MCAO/reperfusion and serial MRI scans; a subset of animals received citalopram from day 7 after MCAO | Male 129/SV mice | Delayed citalopram reversed the behavioral phenotype blocked the degeneration of dopaminergic midbrain neurons, and attenuated striatal atrophy after 4 months (p < 0.05). |
| Natalizumab | ||||
| Becker et al. [213] | 2001 | Rats underwent 3 h of MCAO followed by 45 h of reperfusion. 2 h after ischemia, one group received anti-α4 integrin antibody intraperitoneally and another an isotype control antibody | Male Lewis rats | Neurological deficits were less frequent in treated rats at 24 (p < 0.01) and 48 h (p = 0.01) after ischemia. White blood cell count was higher in treated rats (p < 0.01) with a lymphocyte/monocyte predominance. Infarction volume was reduced in treated animals (p = 0.012). |
| Relton et al. [237] | 2001 | Rats underwent 1-h MCAO followed by 23-h reperfusion. 24 h before MCAO were injected intravenously with anti-α4 integrin antibody (2.5 mg/kg) or isotype control antibody | Male spontaneously hypertensive rats or Sprague-Dawley rats | Treated animals showed reduced total infarct volume (p < 0.05–0.01). Moreover, treatment reduced brain myeloperoxidase activity (p < 0.05). No significant difference in white blood cell count was observed. Leukocyte counts were elevated in TA-2-treated rats. |
| Liesz et al. [226] | 2011 | 24 h before or 3 h after ischemia, mice were administered 300 mg of CD49d-specific monoclonal antibody intraperitoneally after; control animals received rat IgG2b isotype control monoclonal antibody | Male mice C57BL/6J aged 10–12 weeks | VLA-4 blockade improved outcome after 7 days from MCAO via the inhibition of cerebral leukocyte invasion and neurotoxic cytokine production (p < 0.01). VLA-4 inhibition reduced the post-ischemic VCAM-1 up-regulation (p < 0.01). |
| Langhauser et al. [224] | 2014 | 24 h before or 3 h after cerebral ischemia (both tMCAO and pMCAO), mice were treated with 300 μg of a monoclonal antibody anti-CD49d | Male C57Bl/6 mice | VLA-4 blocking reduced T cell and neutrophil invasion after 5 days following MCAO and inhibited the up-regulation of VCAM-1 (p < 0.05). Anti-CD49d antibody could not influence stroke outcome positively, irrespective of the model or the time point investigated. |
| Neumann et al. [232] | 2015 | After focal cerebral ischemia was induced by pMCAO, anti-CD49d treatment was administered intravenously | LysM-eGFP mice | The systemic blockade of VLA-4 resulted in reduction of adherence of neutrophils (p < 0.05) and inhibition of their infiltration (p < 0.01) 24 h after focal ischemia. Moreover, anti-VLA-4 treatment improved neurological outcome and reduced infarct volume at day 3 after stroke (p < 0.05). |
| Llovera et al. [230] | 2015 | After cMCAO (for small lesions confined to the cortex) or fMCAO (for lesions in the cortex and subcortical structures) was assessed, anti-CD49d treatment was administered intraperitoneally 3 h after stroke induction | 315 male C57BL/6J mice | Anti-CD49d treatment reduced infarct volume (p < 0.05) and leukocytes invasion into the ischemic brain (p < 0.001) after 7 days from cMCAO (p < 0.05). After fMCAO, mice had fewer cerebral leukocytes than after cMCAO (p < 0.001), but anti-CD49d treatment did not affect leukocyte invasion after fMCAO. |
| Cyclosporine A | ||||
| Uchino et al. [243] | 1998 | CsA was given intraperitoneally daily for 1 week before and 1 week after forebrain ischemia of 7 or 10 min duration | Rats | Systemically administered CsA ameliorated the damage to the CA1 sector of the hippocampus due to transient ischemia (p < 0.001). |
| Cho et al. [217] | 2013 | Rats underwent MCAO and then randomly treated by intracarotid CsA 10 mg/kg 20 min before MCAO (pre-treatment group); intracarotid CsA 10 mg/kg immediately after reperfusion (post-treatment); and intracarotid saline immediately after reperfusion | 27 Sprague-Dawley rats | On day 1, a significant reduction of infarct size in the pre-treatment group compared to the post-treatment (p < 0.004) was evaluated. A significant reduction of microglial cell count in the pre-treatment group compared to either saline or post-treatment groups was found (p < 0.001). |
| Yu et al. [250] | 2004 | Rats underwent MCAO then were randomly treated with either: low dose CsA, MP, low dose CsA plus MP, high dose CsA, or vehicle | Adult Sprague-Dawley rats | Animals receiving high dose CsA alone exhibited a minor motor asymmetry and less neurologic deficits 3 days after stroke (p < 0.0001) as well as those receiving low dose CsA and MP treatment but only on day 1 post-stroke (p <0.005). Animals receiving high dose CsA alone exhibited significantly (p < 0.0001). |
| Yuen et al. [253] | 2011 | Rats were equally divided into sham control, intraperitoneal physiological saline (at 0.5/24/48 h after stroke), CsA (20 mg/kg at 0.5/24 h intraperitoneally), EPO (5000 IU/kg at 0.5/24/48 h, subcutaneously), combined CsA and EPO after occlusion of distal left internal carotid artery | 50 adult-male Sprague-Dawley rats | On day 21, improvement in neurological function was found in CsA and EPO group (p < 0.05) and was higher when the combined treatment was administered (p < 0.004). Attenuation of inflammatory response, apoptosis, and oxidative stress was found with combined therapy with CsA and EPO (p < 0.05). |
| Edaravone | ||||
| Fujiwara et al. [221] | 2016 | Before 90-min MCAO followed by reperfusion, rats were randomly assigned to intravenous vehicle or intravenous edaravone 3 mg/kg | Male Sprague-Dawley rats | Edaravone decreased infarct volume and edema formation and IL-1β and MMP-9 levels 3 h after ischemia levels (p < 0.05). Edaravone was shown to reduce levels of many other pro-inflammatory cytokines. |
| Yamashita et al. [248] | 2015 | Thrombolysis was evaluated by using a He-Ne-laser-induced thrombosis model in mesenteric microvessels. 3 experimental groups (placebo, alteplase 0.6 mg/kg, alteplase 0.6 mg/kg + edaravone 10.5 mg/kg) | Male Wistar–ST rats | In the alteplase group, thrombus volume decreased (p < 0.01) after 20 min. In the alteplase+edaravone group, thrombus volume was more evident (p < 0.001). |
| Wu et al. [245] | 2014 | Rats were subjected to tMCAO and then administered edaravone 2.4 mg/kg; a subset of these animals were administered both edaravone 2.4 mg/kg and borneol 0.6 mg/kg | Sprague-Dawley rats | Edaravone was demonstrated to scavenge free radicals. Edavarone and borneol reduced the infarct area (p < 0.001) and the effect was increased when drugs were administered synergistically (p < 0.001). |
IL-1Ra: interleukin-1 receptor antagonist; pMCAO: permanent middle cerebral artery occlusion; CV: cardiovascular; PMN: polymorphonuclear; tMCAO: transient middle cerebral artery occlusion; DCP: dorsal area of caudate putamen; VCP: ventral area of the caudate putamen; rIL-1β: recombinant interleukin-1β; MRI: magnetic resonance imaging; MMP: metalloproteinase; CBF: cerebral blood flow; eNOS: endothelial nitric oxide synthase; t-PA: tissue-type plasminogen activator; VEGF: vascular endothelial growth factor; hBMEC: human brain microvascular endothelial cells; OGD: oxygen glucose deprivation; ICH: intracerebral hemorrhage; ICAM-1: intercellular adhesion molecule-1; IFN-γ: interferon-γ; IL: interleukin; I/R: ischemia/reperfusion; CDK5: cyclin-dependent kinase 5; SOD: superoxide dismutase; MDA: malondialdehyde; CaMKII: calmodulin-dependent protein kinase II; CREB: cyclic adenosine monophosphate responsive element binding protein; Kv channels: voltage-gated potassium channels; VCAM-1: vascular cell adhesion molecule-1; VLA-4: very late antigen-4; LysM–EGFP: lysozyme M promoter driving expression of enhanced green fluorescent protein; cMCAO: coagulation of the distal middle cerebral artery; fMCAO: occlusion of the middle cerebral artery with an endovascular filament; CsA: cyclosporine A; MP: methylprednisolone; EPO: erythropoietin.