Study On The Mechanism Of Breviscapine With Exercise Inhibiting Neuronal Autophagy After Stroke

Objective: To record the influence on breviscapine with exercise inhibiting neuronal autophagy after cerebral ischemia reperfusion injury in rats.To investigate the mechanism of breviscapine combined with running platform training to inhibit the neuronal excessive autophagy of rats after stroke.Methods: In adult male Sprague Dawley rats,left middle cerebral artery occlusion(MCAO)model was prepared and randomly divided into the following groups:sham-operated group(Sham),saline group(MCAO+Saline),running platform training group after MCAO(MCAO+Exercise),breviscapine low-dose group(MCAO+0.1mg/ml Bre),breviscapine medium-dose group(MCAO+0.2mg/ml Bre),breviscapine high-dose group(MCAO+0.4mg/ml Bre)and breviscapine with running platform training group(MCAO+0.1mg/ml Bre+Exercise).Rats in Sham group and MCAO+Saline group were not exercised,whereas rats in groups of MCAO+Exercise and MCAO+0.1mg/ml Bre+Exercise were involved in running training with gradually increasing intensity(30min per day for 7 days).At 1,3 and 7 days after cerebral ischemia-reperfusion injury,behavioral methods such as Foot-fault test,neurological deficit score and rod fatigue test were used to evaluate the recovery of neurological function and improvement of motor function.At the 7th day,the neuronal damage in the penumbra of cerebral infarction was detected by cresyl violet staining.Western Blot was applied to detect the expression levels of autophagy related proteins on day 3 and 7,whereas immunofluorescence was used to locate autophagy and apoptotic cells on day 7.Results:(1)The optimal dose of breviscapine(0.1mg/ml Bre)was determined as the intervention dose for the subsequent experiment.(2)Compared with the MCAO+Saline group,the symptoms of neurologic loss were significantly alleviated in the MCAO+Exercise group,MCAO+0.1mg/ml Bre group and MCAO+0.1mg/ml Bre+Exercise group.MCAO+Exercise group improved the recovery of neurological function comparing with MCAO+0.1mg/ml Bre and MCAO+0.1mg/ml Bre+Exercise group,suggesting that the combination group did not necessarily have a better effect.(3)Western Blot and immunofluorescence results showed that breviscapine combined with running platform training significantly inhibited the expression of autophagy-related proteins in neurons.(4)Brevicapine combined with exercise training inhibited the expression of autophagy-related proteins,and the expression of JNK2 protein was also decreased suggesting that the inhibition of excessive autophagy may be through the regulation of JNK pathways.Conclusion:(1)Breviscapine combined with running platform training after cerebral ischemia can reduce cerebral infarction volume and promote the improvement of neurological function in rats.(2)Breviscapine combined with running platform training after cerebral ischemia can inhibit the expression of autophagy-related proteins in neurons.(3)We concluded that breviscapine combined with running platform training inhibited the expression of LC3 B and Beclin1 proteins,which may inhibit neuronal over-autophagy and promote the protection of neurons after brain injury by regulating JNK-related pathways.