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Objective To screen protein disulfide isomerase(PDI)inhibitors from zinc- and selenium-rich green tea and to examine the effect of PDI inhibitors on platelet aggregation. Methods Protein structure file of PDI(PDB ID:4EKZ)was downloaded from protein data bank and the current known 20 compounds in green tea were used to establish a small chemical library. The effects of 6 hit compounds by virtual screening on enzymatic activity of PDI were validated. The antiplatelet activities of the effective compounds tested on PDI enzymatic activi ty were further evaluated. Results 2 of 6 hit compounds by virtual screening ,ECG and EGCG displayed inhibitory effect on enzymatic activity of PDI. In addition ,both compounds showed the potential inhibitory effect on platelet aggregation induced by adenosine diphosphate (ADP) in vitro. Conclusion The effect of blood-activating and stasis-dissolving regulation of green tea is associated with its inhibitory effect on PDI. ECG and EGCG are major active components.
RESUMO
Objective To screen protein disulfide isomerase(PDI)inhibitors from zinc- and selenium-rich green tea and to examine the effect of PDI inhibitors on platelet aggregation. Methods Protein structure file of PDI(PDB ID:4EKZ)was downloaded from protein data bank and the current known 20 compounds in green tea were used to establish a small chemical library. The effects of 6 hit compounds by virtual screening on enzymatic activity of PDI were validated. The antiplatelet activities of the effective compounds tested on PDI enzymatic activi ty were further evaluated. Results 2 of 6 hit compounds by virtual screening ,ECG and EGCG displayed inhibitory effect on enzymatic activity of PDI. In addition ,both compounds showed the potential inhibitory effect on platelet aggregation induced by adenosine diphosphate (ADP) in vitro. Conclusion The effect of blood-activating and stasis-dissolving regulation of green tea is associated with its inhibitory effect on PDI. ECG and EGCG are major active components.
RESUMO
Aim Toinvestigatethepro-angiogenic effects of Danshensu derivative ADTM and explore its underlying possible signaling pathway using zebrafish embryosasinvivomodels.Methods Theangiogenesis activities of ADTM were determined in experimental models of normal and VEGFR tyrosine kinase inhibitorⅡ(VRI )-induced vascular defective zebrafish embry-os.Embryos were treated with various concentrations (50,100,200 μmol · L-1 ) of ADTM for indicated time.The diameter and the numbers of endothelial cells of zebrafish SIVs were evaluated,respectively.In VRI model,the number of intact and defective ISVs in each zebrafish embryo was counted.The total RNA of zebrafish embryos was extracted and transcriptional profiling was analyzed by deep sequencing.Quantita-tive real-time PCR(qPCR)was performed to 4 genes selected from transcriptional profiling to validate the data collected from transcriptome analysis.Results ADTMsignificantlyincreasedsubintestinalvessels (SIVs)diameter in a concentration-dependent manner in normal zebrafish as well as restored VRI-induced blood vessels defect in VRI-exposed zebrafish. The transcriptome data analysis demonstrated that 19 signif-icantly changed genes were mapped to insulin signaling pathway.The qPCR data are in good agreement with those obtained by deep sequencing and support the consistency between the two methods for determining relative expression levels in the zebrafish model.Con-clusion Inzebrafishmodel,ADTMexhibitsthe effects of angiogenesis and blood vessel restoration. The underlying mechanism may be involved in the acti-vation of insulin signaling pathway.