Inhibitory effect of Edaravone Dexborneol on neuroinflammation in ischemic stroke rats and its mechanism
Objective To investigate the therapeutic effect of Edaravone Dexborneol(EDB)on ischemic stroke and explore its under-lying mechanism.Methods Sprague-Dawley rats were divided into four groups:sham group,MCAO group,EDB group(MCAO+EDB 2.5 mg/kg),and Edaravone group(MCAO+Edaravone 2.5 mg/kg).Middle cerebral artery occlusion(MCAO)was employed to induce the ischemic stroke(IS)rat model in model group and drug administration group.The rats in sham operation group were treated with the same procedure as model group except no thread thrombus.The neurological severity score was used to assess the neurological deficits.2,3,5-triphenyltetrazolium chloride(TTC)staining was used to assess the cerebral infarction.ELISA assay was used to detect the levels of inflammatory factors in brain tissues.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to examine the expressions of chemokines(CCL2,CCL5,CCR5,CCL7,CXCL1,CXCL2,and CXCL12),TLR4 and NF-κB.In vitro,BV2 cells were divided into six groups:control group,lipopolysaccharide(LPS)group,low,medium and high dose EDB groups(5,10,50 μg/mL),and Edaravone group(10 μg/mL).BV2 cells were stimulated with 10 μg/mL LPS for 24 h to simulate the neuroinflammatory state.The production of NO in cellular supernatants was detected by NO assay kit.The mRNA expressions of CCL2,TLR4,and NF-κB were determined by RT-qPCR.Protein expressions of CCL2,TLR4,NF-κB and p-NF-κB were determined by Western blot.Results Compared to model group,the neural function was improved in EDB group(P＜0.01),and the cerebral infarction area and the levels of inflammatory factors were significantly decreased(P＜0.01).Compared with Edaravone group,the neurological severity score,and the infarct volume and the inflammatory factors were significantly decreased in EDB group(P＜0.05).Compared to sham group,the expressions of chemokines CCL2,CXCL12,CCL7,CXCL2,CCL5 and CXCL1 in model group were significantly increased,especially CCL2(P＜0.01).Compared to model group,the mRNA expressions of CCL2,CXCL12,CXCL1,CXCL2 and CCL5 were significantly decreased in EDB group(P＜0.01).Compared with Edaravone group,the mRNA expression level of CCL2 was decreased in EDB group(P＜0.05).Compared with model group,the mRNA expressions of CCL2,TLR4 and NF-κB in EDB group were significantly decreased(P＜0.01).Compared to Edaravone group,the mRNA expression levels of CCL2 and NF-κB in brain tissue were significantly decreased in EDB group(P＜0.05).Compared to LPS group,the NO content was significantly decreased in EDB groups(P＜0.01),especially in high-dose group,and the mRNA and protein expressions of CCL2,TLR4 and NF-κB were significantly decreased(P＜0.01).Compared to Edaravone group,the mRNA expression levels of CCL2 and TLR4 were significantly decreased in EDB groups(P＜0.05),and the expression levels of TLR4 and NF-κB protein were also significantly decreased(P＜0.05).Conclusion EDB can exert a protective effect against neuroinflammation in ischemic stroke rats by inhibiting the CCL2-NF-κB signaling pathway.
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