Effect of Buyang Huanwu decoction combined with bone marrow mesenchymal stem cell transplantation on expression of integrin in a rat model of middle cerebral artery occlusion / 中国组织工程研究

ABSTRACT

BACKGROUND:Previous studies have found that combined use of Buyang Huanwu decoction and bone marrow mesenchymal stem cel (BMSC) transplantation can play a synergic role against cerebral ischemia injury. OBJECTIVE:To analyze the effect of Buyang Huanwu decoction combined with BMSC transplantation to promote angiogenesis after cerebral ischemia. METHODS:Ninety-six Sprague-Dawley rats were randomly divided into four groups, and used to make middle cerebral artery occlusion models. In combined group, rats were given intragastrical administration of Buyang Huanwu decoction 10 mL/kg once a day, beginning at 3 days prior to modeling, and then given intragastrical administration of Buyang Huanwu decoction once at 2 hours after modeling, fol owed by its intragastrical administration every 12 hours. In BMSC and combined groups, BMSC suspension was injected into the rat ventricle after 2-hour cerebral ischemia/2-hour reperfusion, and then 30 minutes later, CD34 and CD45 antibodies were injected. In antibody group, CD34 and CD45 antibodies were injected. In model group, only normal saline was given. SerumαVβ3 level detection, immunohistochemical observation, Q-PCR and western blot tests were performed in the combined group at 12, 24, 36, 48 hours after reperfusion, while these indices were detected in the other three groups at 36 hours after reperfusion. RESULTS AND CONCLUSION:(1) The level of serumαVβ3 was lower in the antibody group than the model group (P<0.05), higher in the BMSC and combined groups than the antibody group (P<0.05), and higher in the combined group than the BMSC group (P<0.05). (2) Immunohistochemical findings showed that compared with the antibody group, the number of CD34 positive cel s was higher in the model, BMSC and combined groups (P<0.05). (3) Results from the Q-PCR and western blot assay showed that compared with the model group, the pFAK protein expression level was lower in the antibody and BMSC group, but FAK gene expression level had no overt changes;while the protein levels of FAK (24 hours after reperfusion) and pFAK (12 hours after reperfusion) were significantly increased in the combined group than the antibody and BMSC groups (P<0.05, P<0.01). Moreover, this increase exhibited a gradual y rising trend with the extension of reperfusion time. To conclude, the combined use of BMSC transplantation and Buyang Huanwu decoction can reverse the effect of CD34+CD45 antibodies that lead to the decrease in the number of vascular endothelial cel s and levels of integrinαVβ3 and downstream signaling molecules, thereby to promote angiogenesis in the MCAO model.