| BackgroudThe occurrence and development of cerebral ischemia-reperfusion injury is a complex pathophysiological process.Among them,oxidative stress is closely related to neuronal death following cerebral ischemia-reperfusion injury.Studies have found that the activation of endogenous antioxidant system plays a crucial role in the survival of nerve cells after cerebral ischemia-reperfusion injury.Dj-1(also known as PARK7,or PARK7)is one of the multifunctional proteins selected through evolution.It plays an important role in anti-apoptosis,mediating cell signal transduction and resisting oxidative stress in the body.Recently,it has been found that Dj-1 plays an important role in protecting nerve cells by relying on its good antioxidant effect.Dj-1 is closely related to the pathogenesis of Parkinson’s disease.Dj-1 can protect neurons from oxidative stress damage in Parkinson’s disease.Recently,the protective effect of DJ-1 in ischemic stroke has been extensively studied,but the specific mechanism remains unclear.Transcription factor NF-E2-related factor 2,a member of the transcription factor family,acts as a master regulator of the body’s antioxidant defense system through binding with antioxidant response elements(ARE).As an antioxidant protein,dj-1 plays an important role in the regulation of Nrf2 activity.However,whether DJ-1 promotes the nuclear transfer of Nrf2 and activates its antioxidant function by upregulation of Nrf2 or by regulating the subcellular localization of Nrf2 has not been reported yet.ObjectiveThe purpose of this study was to explore the role of DJ-1 in cerebral ischemia-reperfusion oxidative stress injury and its related regulatory mechanism,so as to provide theoretical basis for further prevention and treatment of cerebral ischemia-reperfusion oxidative stress injury.Methods(1)In this study,male adult SD rats were selected as research subjects,and Middle Cerebral Artery Occlusion(MCAO)model was established to simulate Cerebral ischemia reperfusion(I/R)injury in rats.(2)DJ-1 si RNA was injected into the left ventricle 24 h before MCAO/R,to observe neurological functional scores cerebral water content and the effect of brain tissue morphology in DJ-1 si RNA group after MCAO.(3)To analyze the effects of interference of DJ-1 expression on oxidative stress level after MCAO/R,superoxide dismutase(SOD)and malondiadehyde(MDA)were measured by ELISA.(4)Western Blot was used to analyze the effect of interference of DJ-1on the expression of DJ-1,Nrf2,HO-1 and NQO1 in brain tissue of rats after MCAO/R.(5)Adeno-Associated Virus(AAV)of Dj-1 overexpression was injected into the left cerebral cortex of SD rats one month before MCAO/R.(6)Western Blot was used to analyze the effect of overexpression of DJ-1 on the expression of DJ-1,Nrf2,HO-1 and NQO1 in brain tissue of rats after MCAO/R.(7)Immunofluorescence staining was used to observe expression and nucleation of Nrf2 after MCAO/R.Results(1)Compared with MCAO group,the neurofunctional scores and brain water content of DJ-1 si RNA group were significantly increased.HE and Nissl staining showed further aggravation of neuronal injury in cerebral ischemia area.SOD content was further reduced and MDA content was further increased.(2)After interference with DJ-1,the protein level of Nrf2 and its downstream HO-1 and NQO1 were also significantly reduced.(3)After overexpression of DJ-1,the protein level of DJ-1,Nrf2 and their downstream HO-1 and NQO1 were significantly increased.ConclusionAs an important neuroprotective factor in vivo,DJ-1 can reduce oxidative stress injury caused by cerebral ischemia reperfusion in rats,which may be realized by activating Nrf2 signaling pathway. |
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