ABSTRACT
Objective To explore the effect of astragalus extracts on adherence, migration, cell viability, microvascular formation, and eNOS expression in bone marrow-derived endothelial progenitor cells (EPCs) of rats. Methods EPCs were cultured, isolated and identified in vitro and divided into four groups: low titer group (10-4 g/L of astragalus extracts), middle titer group (10-3 g/L of astragalus extracts), high titer group (10-2 g/L of astragalus extracts) and control group. The effects of astragalus extract on adhesion, migration or microvascular formation were observed under the inverted microscope and com?pared between all groups;Cell viability was detected by MTT assay;mRNA transcription and protein expression levels of en?dothelial nitric oxide synthase (eNOS) in EPCs were detected by reverse transcription PCR (RT-PCR) and Western blot re?spectively. Results Compared with the control group, cell adhesion, migration and microvascular formation of EPCs all en?hanced with addition of astragalus extracts in a dose dependent manner (F=15.256, 13.633, 97.549 respectively, and P <0.05 in all cases); Cell vitality of EPCs increased with administration of astragalus extracts in a time and dose dependant manner. (F=9.755 for time and F=10.18 for dose). mRNA transcription and protein expression of eNOS in EPCs were up-reg?ulated with addition of astragalus extracts in a dose dependent manner (F=56.356 and 77.125 respectively, and P<0.05 in both cases). Conclusion Astragalus extracts play important role in angiogenesis in EPCs probably through up-regulating expressions of eNOS.
ABSTRACT
Aim To explore the effect of protein kinase D1 (PKD1 )on adherence,migration,proliferation, microvascular formation,and eNOS expression in bone marrow-derived endothelial progenitor cells(EPCs)of rats.Method The EPCs were isolated from bone marrow of rats,cultured and detected,the effects of PKD1 and its specific blocking agent CID755673 on adhesion,migration,proliferation,or microvascular formation were observed,as well as mRNA and protein expression of endothelial nitric oxide synthase (eNOS ) in EPCs.Results PKD1 significantly promoted adhe-sion,migration,proliferation and microvascular forma-tion of EPCs,and upregulated the mRNA and protein expression of eNOS in EPCs,according to the cell cul-ture experiments of EPCs in vitro.Conclusion PKD1 has the role of angiogenesis through regulation of EPCs,which might be dependent on eNOS.
ABSTRACT
AIM:Todeterminetheeffectofsalviaextractonangiogenesisofthemyocardiumintheratswith myocardial infarction (MI) and to analyze its possible mechanism .METHODS: Left coronary artery of Sprague-Dawley rats was ligated to establish a MI model .The rats were randomly divided into MI model group , 3 different dose groups of salvia (10, 20 and 40 mg? kg-1? d-1), and sham operation group.Each group consisted of 8 rats.The rats in all treat-ment groups were orally administered with the salvia extract , and the rats in MI group and sham operation group were fed with the same volume of saline .The rats were sacrificed 4 weeks later .The hemodynamic changes of the rats were deter-mined , and the segmental heart samples were used for morphological observation by hematoxylin and eosin staining , Masson staining, or electron microscopic analysis.The expression of vascular endothelial growth factor (VEGF) and cluster of dif-ferentiation 34 ( CD34 ) was analyzed according to immunohistochemistry .RESULTS: Compared with sham operation group, the morphological changes of the myocardium in MI group were disordered , part of myocardial cell outline disap-peared , and obvious fibrosis in the necrosis myocardial tissue and fuzzy or disappearing microvascular ultrastructure were al -so observed .Compared with MI group , the number of new microvessels in all the treatment groups increased obviously , and the morphological changes of the endothelial cells were relatively complete according to electron microscopy .Compared with sham operation group , the protein expression of VEGF and CD 34 in the cytoplasm of the myocardial tissues in MI group in-creased only a little .Compared with MI group , the protein expression of VEGF and CD 34 in the cytoplasm of the myocardi-al tissues in all treatment groups increased significantly ( P<0.01 ) .CONCLUSION: Salvia extract obviously promotes angiogenesis of the myocardial tissues in the rats after myocardial infarction .