Abstract:
BACKGROUND: Buqi Huoxue Compounds have significant clinical efficacy in treating ischemic stroke with Qi deficiency and phlegm stasis; however, the exact mechanism of action is not clear.
OBJECTIVE: To observe the effect of Buqi Huoxue Compounds on the expression of vascular endothelial growth factor, basic fibroblast growth factor, brain-derived neurotrophic factor and autophagy related protein Beclin1 and p62 in a rat model of cerebral ischemia/reperfusion.
METHODS: Forty male Sprague-Dawley rats were randomly divided into sham operation group, model group, Buqi Huoxue Compounds group and autophagy inhibitor group, with 10 rats in each group. In the latter three groups, a rat model of cerebral ischemia/reperfusion injury was established. The Buqi Huoxue Compounds group was intragastrically given Buqi Huoxue Compounds (6.49 g/kg, administered three times a day) 2 hours after reperfusion; the autophagy inhibitor group was intragastrically given Buqi Huoxue Compounds (6.49 g/kg, administered three times a day) 2 hours after reperfusion and intraperitoneally given 3-methyladenine 2 hours before gavage and at days 1-3 of gavage. The sham operation group and model group were given equal amounts of saline by gavage for 7 consecutive days. Neurological function, cerebral infarct volume, brain tissue morphology and expression of vascular endothelial growth factor, basic fibroblast growth factor, brain-derived neurotrophic factor and autophagy-related proteins Beclin1 and p62 in the ischemic cortical region of rats were detected at 24 hours after the final administration.
RESULTS AND CONCLUSION: Zea-Longa scoring results showed that the neurological function of rats was severely damaged after modeling and neurological deficit of rats in the Buqi Huoxue Compounds group was less than that in the model group and the autophagy inhibitor group (P < 0.05). TTC staining showed that cerebral infarct foci were observed in the model group, Buqi Huoxue Compounds group, and autophagy inhibitor group, and the cerebral infarct volume in the Buqi Huoxue Compounds group was lower than that in the model group and the autophagy inhibitor group (P < 0.05). The results of hematoxylin-eosin staining in ischemic brain tissues showed that there were large gaps between nerve cells in the model group and cell arrangement was not neat, and cytoplasmic agglutination and pyknosis were observed. Immunohistochemical staining results showed that vascular endothelial growth factor was mostly expressed in neuronal cells, glial cells and capillary endothelium; basic fibroblast growth factor and brain-derived neurotrophic factor were mostly expressed in neuronal cells and glial cells; and there was no significant difference in the expression of vascular endothelial growth factor, basic fibroblast growth factor, and brain-derived neurotrophic factor among the four groups (P > 0.05). The results of western blot assay showed that compared with the sham operation group, Beclin1 protein expression was decreased (P < 0.05) and p62 protein expression was elevated (P < 0.05) in the model group; compared with the model group, Beclin1 protein expression was increased (P < 0.05) and p62 protein expression was reduced (P < 0.05) in the Buqi Huoxue Compounds group; compared with the Buqi Huoxue Compounds group, Beclin1 protein expression was decreased (P < 0.05) and p62 protein expression was elevated (P < 0.05) in the autophagy inhibitor group. To conclude, Buqi Huoxue Compounds attenuate cerebral ischemia-reperfusion injury in rats by promoting autophagy.

Key words: Buqi Huoxue Compounds, ischemia/reperfusion, neurologic deficit, autophagy, vascular endothelial growth factor, basic fibroblast growth factor, brain-derived neurotrophic factor, Beclin1, p62