ABSTRACT
AIM:To investigate the protective effect of Huoxue-Dingxuan capsule-medicated serum on mouse brain microvascular endothelial cell line bEnd .3 with hypoxic injury .METHODS:The Wistar rats were randomly divided into control group and Huoxue-Dingxuan capsule group .The serum was collected at the 7th day after drug administration . The bEnd.3 cells were divided into normal group , hypoxia serum model group and Huoxue-Dingxuan capsule serum group . After administration of the medicated serum , bEnd.3 cell were treated with hypoxia for 6 h.The cell morphology was ob-served under microscope , the apoptosis rate and cell cycle were detected by flow cytometry , and the activity of superoxide dismutase ( SOD) and the content of malondialdehyde ( MDA) were detected by ELISA .RESULTS:Compared with other groups, Huoxue-Dingxuan capsule-medicated serum significantly resisted the injury caused by hypoxia , obviously improved the morphology of bEnd .3 cells, significantly reduced the number of apoptotic cells induced by hypoxia , and effectively in-hibited the occurrence of hypoxia-induced G1/S phase arrest in the bEnd.3 cells.At the same time, the medicated serum inhibited MDA production , and increased SOD activity .CONCLUSION: Huoxue-Dingxuan capsule attenuates hypoxia-induced bEnd .3 cell damage by enhancing the antioxidant capacity of cells and inhibiting cell apoptosis .
ABSTRACT
Objective: To investigate the effect of Huoxue Dingxuan Capsule (HDC) containing serum on gene expression profile in mice brain microvascular endothelial cell bEnd.3 and its molecular mechanism. Methods: Thirty rats were randomly divided into control group and HDC group, each group had 15 rats, and the serum was collected in two groups of rats. BEnd.3 was divided into blank serum group, hypoxia model group, and 10% HDC containing serum group. The cells were intervened in different conditions, and after oxygen deprivation for 6 h, the cytoskeleton was observed under laser microscope. And to detect the effect of HDC containing serum on gene expression profile in mice brain microvascular endothelial cell bEnd.3 by Affymetrix U133 plus2.0 gene expression microarray, and for clustering analysis by molecular annotation system MAS3.0. Results: Differentially expressed genes were identified based on P 3. There were 405 differentially expressed genes between blank group and model group, in which 176 genes were up-regulated and 229 genes were down-regulated. There were 368 differentially expressed genes between HDC sero group and model group, in which 146 genes were up-regulated and 222 genes were down-regulated. These differentially expressed genes had the functions of positive regulation of endothelial cell migration, process of acid metabolism, production of vascular endothelial growth factors, positive regulate activity of NF-κB transcription factors, oxidative stress-induced senescence, mitosis prophase, platelet aggregation (blockage formation), ect. And many signaling pathways were regulated by these genes including vascular endothelial growth factor signaling pathway, interleukin signal pathway, p53 pathway, TNF-signal pathway, PPAR signaling pathway, PI3K-Akt signaling pathway, apoptosis signal pathway etc. Conclusion: HDC has significant inhibitory effects on damage of bEnd.3 cells induced by hypoxia. Its mechanism is related to oxidative stress, inhibition of apoptosis, promotion of angiogenesis, inflammation, and immune response. Huoxue Dingxuan Capsule can regulate the function of vascular endothelial cell on gene level by several signaling pathways.