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Objective:To investigate the neuroprotective effect of long-term prophylactic use of buphthalein on mice with permanent distal middle cerebral artery occlusion and its relationship with the nuclear factor erysid 2 related factor 2 (Nrf2) pathway.Methods:Nrf2 + /+ wild-type and Nrf2 -/- knockout mice were randomly divided into control group (equal volume vegetable oil), low-dose butylphthalide group (20 mg/kg) and high-dose butylphthalide group (60 mg/kg), with 6 mice in each group. The drug was administered once a day by gavage for 1 month, and then a permanent middle cerebral artery occlusion model was induced by electrocoagulation. After the model was made, the drug was continued and the mice were sacrificed on the 10 th day. The modified Longa grading scale and the rotating rod test were used to evaluate neurological deficits on the 3 rd and 10 th day after the model was made. After the mice were sacrificed, the cerebral infarct volume was measured by triphenyltetrazolium chloride staining. The brain water content was measured by dry and wet weight method. The expression of Nrf2 pathway related factors, including Nrf2, heme oxygenase 1 (HO-1) and NAD(P)H quinone oxidoreductase 1 (NQO1) were measured by quantitative real-time PCR and Western blotting. Results:On the 10 th day after modeling, compared with the Nrf2 -/- control group, the neurological deficit was significantly milder, the volume of cerebral infarction and brain water content were significantly smaller, and the mRNA and protein levels of Nrf2, HO-1 and NQO1 were significantly higher in the Nrf2 + /+ control group, and the differences were statistically significant ( P<0.05). For Nrf2 + /+ mice, compared with the control group, the cerebral infarct volume was significantly reduced ( P<0.05), the brain water content was significantly reduced ( P<0.05), and the neurological function recovery was significantly better ( P<0.05), and the levels of Nrf2, HO-1, and NQO1 mRNA and protein were significantly higher in the high-dose butylphthalide group (all P<0.05). For Nrf2 -/- mice, there were no significant differences in neurological function, cerebral infarction group volume, brain water content, Nrf2, HO-1, NQO1 mRNA and protein levels among the groups. Conclusion:Long-term butylphthalide pretreatment can significantly improve the neurological function, reduce cerebral infarction volume, reduce brain water content, and increase Nrf2, HO-1, NQO1 mRNA and protein expression levels in mice with permanent distal middle cerebral artery occlusion, suggesting butylphthalide may play a neuroprotective effect by up-regulating the expression of Nrf2 gene and its downstream antioxidant stress factors HO-1 and NQO1.
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Objective To investigate the correlation between plasma cathepsin L (CatL) levels and establishment of cerebral collateral circulation in acute ischemic stroke patients with cerebral artery stenosis.Methods The patients with acute ischemic stroke with at least one cerebral large artery (including internal carotid artery,middle cerebral artery,vertebral artery,and basilar artery) stenosis ＞ 70％ diagnosed by whole cerebral angiography were enrolled.ASITN/SIR blood flow classification system was used to systematically evaluate the establishment of cerebral collateral circulation.Grade 0-2 was defined as poor collateral branch and 3-4 was defined as good collateral branch,Enzyme linked immunosorbent assay was used to detect the plasma CatL level.Results A total of 79 acute ischemic stroke patients with cerebral artery stenosis were enrolled,including 63 male and 16 female.Their mean age was 58.76 ± 12.24 years old.There were 51 patients (64.56％) in the poor collateral branch group and 28 (35.44％) in the good collateral branch group.There was no significant difference in plasma CatL levels between the good collateral circulation group and the poor collateral circulation group (7.09± 2.27 mg/L vs.8.79±3.53 mg/L;t =2.751,P =0.069).Multivariable logistic regression analysis showed that only the high National Institutes of Health Stroke Scale score was the independent risk factor for poor collateral circulation (odds ratio 0.935,95％ confidence interval 0.823-0.963;P=0.046),and there was no significant independent correlation between plasma CatL levels and collateral circulation (odds ratio 0.910,95％ confidence interval 0.766-1.081;P =0.285).Conclusion There was no significant correlation between plasma CatL levels and cerebral collateral development in acute ischemic stroke patients with cerebral artery stenosis.