Effect of Xuefu Zhuyu Capsule on Angiogenesis in Hindlimb Ischemic Rats

Abstract

Objective To investigate the effect and mechanism of Xuefu Zhuyu Capsule (XZC) on pro-angiogenesis in the hindlimb ischemic model rats. Methods A total of 100 Sprague Dawley rats were randomly divided into a model group, a regular-dose XZC group (0.48 g center dot kg(-1)center dot d(-1)) and a high-dose XZC group (0.96 g center dot kg(-1)center dot d(-1)) using random number table method. The model of hindlimb ischemic rats were made through femoral artery embolization with Bletilla microsphere agent. XZC were given on the first day after embolization surgery and lasted 5 days. Finally 72 models were obtained with 12 in each group for each time point. The lower ischemic limb was amputated on the third day after embolization surgery. Histopathological characters and the number of blood vessels of granulation tissues were observed at 36 and 48 h after amputation, respectively. The main genes were obtained from microarray analysis and were validated using real-time quantitative polymerase chain reaction. Results The vascular number of granulation tissues at both 36 and 48 h were characterized by new and fresh vessels. The number of angiogenesis in the high-dose XZC group at 36 and 48 h was greater compared with that in the regular-dose XZC and model groups (P < 0.01), and high-dose XZC at 36 h increased more vessels than that at 48 h (P < 0.01). Consequently, granulation tissues from the high-dose XZC group at 36 h were chosen for microarray analysis. In all, 2,085 differentially expressed genes (DEGs) were detected and 25 DEGs were determined to be directly related to angiogenesis. Four biological process terms were found including angiogenesis, regulation of angiogenesis, positive regulation of angiogenesis, and positive regulation of vascular endothelial growth factor receptor signaling pathway (P < 0.05). Microarray analysis also showed 49 pathways including 11 pathways related to angiogenesis. Conclusion XZC promoted angiogenesis moderately and the mechanism involved multiple DEGs and multiple pathways.