EGCG Modulates The Rhythm Of Clock Genes In Vascular Smooth Muscle Cells Induced By Ang?

Circadian clock system plays a critical role in human health.Disorders of this system result in several diseases,including insomnia,hypoimmunity,metabolic syndrome and cancer.In addition,one of important biomarkers of healthy human is the rhythmic blood pressure during day and night.Vascular smooth muscle manages the contraction and relaxation of blood vessels,and thus regulates blood pressure.Therefore,investigating the biological rhythm of vascular smooth muscle cells(VSMCs)is important for homeostasis of blood pressure and health.Based on these factors,this study we successfully set up a circadian rhythm gene expression model of vascular smooth muscle cell induced by Ang ?.Based on this model,qPCR,western blot,transcriptome sequencing and biorhythm fluorescence detection were employed to investigate the EGCG modulating circadian genes expression of VSMCs induced by Ang ?.The main results of this study are shown as follows:The qPCR results demonstrated that EGCG significantly reduced the amplitude of the four circadian genes,which are Bmall,Per2,Cryl and Rev-erba.Meanwhile,EGCG prolonged the periods and delayed the phases.Western blotting results showed that EGCG significantly prolonged the cycle of BMAL1 and REV-ERBa protein rhythmic expression,and down-regulated the amplitudes and delayed the phasesBased on the transcriptome sequencing data,analysis the KEGG signalling pathway of DEGs revealed that EGCG down-regulated the expression of differentially expression genes in circadian rhythm signal pathways,except for Dec]and Dec2.The results suggested that the potential mechanisms of EGCG-regulated rhythm of circadian genes is to stimulate the over expression of Dec1 and Dec2,and to down-regulate of the transcriptional activation of the BMAL1-CLOCK heterodimer by the corresponding protein,consequently regulating other circadian genes.The aorta and mesenteric artery isolated from the PER2::LUC mice undergo rhythm fluorescence detection.The results showed that EGCG reduced the amplitute of the PER2 expression and also delayed the rhythm phase...