| ObjectiveTo explore the activation rule of bcl-2 homologous domain protein(Beclin-1),autophagy-related gene 7(ATG7),autophagy-related gene 3(ATG3)and microtubule-associated protein light chain 3(LC3)in the cerebral cortex after focal cerebral ischemia reperfusion(IR).MethodsHealthy male SD rats were randomly divided into sham operation group(sham group)and model group(IR group).The model group was divided into six subgroups: IR 0h,1h,3h,6h,12 h and 24 h.The middle cerebral artery occlusion(MCAO)was used to establish focal cerebral ischemia reperfusion model.Neurological function score after animals woke up,2,3,5-triphenyltetrazolium chloride(TTC)staining and the water content of brain tissue after 24 h of modeling were used to identify whether the models were successful.Transmission electron microscopy was used to observe the autophagy of neurons in each group.The expressions of Beclin-1,ATG7,ATG3 and LC3 were detected by immunohistochemistry and Western blot.ResultsThe results of model identification showed that the behavior of sham group rats were normal and the symptoms of neurological deficits were not seen.The edema and infarct zone of brain tissue were not seen.The neurological function scores of IR rats showed obvious defects,and the water content of brain tissue was significantly increased after IR 12 h.The obvious pale infarct area was visible after IR 6h.The results were significantly different from those of sham group.By transmission electron microscopy sham group normal neuronal cells were observed and autophagosomes were occasionally observed while the autophagy in neuronal calls of IR groups was significantly observed.After the immunohistochemical staining,a small amount expression of Beclin-1 positive cells in sham group was seen,while the expressions of IR groups were different up-regulated at different time points,and IR 3h and IR 24 h group showed higher expression levels.The same level of expression was also showed through Western blot.And immunohistochemical staining and western blot results showed that expression of ATG7,ATG3,LC3 was consistent with that of Beclin-1.ConclusionsAutophagy-related genes Beclin-1,ATG7,ATG3 and LC3 play an important role in cells autophagy of IR. |
