The Study Of Machanism Effect Of Tianma Gouteng Decoction To Cell Circadian Rhythm When It Regulate The Expression Of Clock Genes

ObjectivesBecause Tianma Gouteng Decoction has the effect of calming liver wind?heat-clearing and blood-activating therapy?Liver and kidney tonic,it is the Clinical classic representative of prescriptions which is used to treat hypertension.There are lots of literatures and reports about using Tianma Gouteng Decoction to treat hypertension.The prophase of this experiment will study the effect of Tianma Gouteng Decoction to the rat model with hypertension,and detect the circadian changes in blood pressure of rats by using the technology of telemetry,and detect the clock genes' expression of each point of time in the aortic of rats.And the result is the Tianma Gouteng Decoction has the adjustment to the circadian blood pressure of rats with hypertension,and can improve the blood pressure rhythm in rats,but it is unclear about the specific mechanism of action and targets.The article is combined with previous studies to clarify the adjustment of Tianma Gouteng and its active ingredients to the possible mechanism of action and targets of circadian blood pressure by examining the intervention effect of Tianma Gouteng Decoction and its active ingredient to clock gene expression of vitro cultured vascular smooth muscle cells(A7r5 cells)and to Ang ?-AT1 downstream pathway proteins.During experiment,through Tianma Gouteng Decoction and its active ingredient to the expression of Ang ? clock genes on each point of time in A7r5 cells and the expression of circadian rhythm to Bmall,Clock,Per2 and mRNA,it demonstrates that Tianma Gouteng Decoction and its effective ingredients adjust the possible mechanism and effect point for circadian rhythm of expression for clock genes.1.Research for Tianma Gouteng Decoction and its effective ingredients adjusting circadian rhythm of expression for clock gene.(1)Effects for Tianma Gouteng Decoction and its effective ingredients for expression of circadian rhythm for clock gene mRAN.Through experiment of Tianma Gouteng Decoction and its effective ingredients to cell A7r5 induced by Ang?,we take method of RT-PCR to research changes for expression of circadian rhythm for clock gene which demonstrates that Tianma Gouteng Decoction and its effective ingredients can make changes for expression of Bmall,Clock and gege mRNA of Per2.The similar change for normal cells of high in day and low in night is shown.(2)The effective ingredients of Tianma Gouteng Decoction affect expression of circadian rhythm for clock gene of cell A7r5 disposed by anticholinergic ATI.After using receptor blocking pharmacon AT1,the expression of circadian rhythm in A7r5 managed by Isorhynchophylline for clock gene in Bmall,Clock and Per2 is reduced.The result demonstrates that Isorhynchophylline adjusts gene Bmall pattern oscillation by intervening receptor AT1 so that cell changes patterns during day and night.(3)Effects of expression for circadian rhythm by Tianma Gouteng Decoction and its effective ingredients to receptor mRNA of AT1Through Tianma Gouteng Decoction and cell A7r5 induced by Ang? from effective ingredients,the regular changes of expression of circadian rhythm for clock gene patterns are shown which are possibly made by intervening receptor Ang?-AT1.2.Research for expression point of circadian rhythm for clock gene by Tianma Gouteng Decoction and its effective ingredients.(1)The intervening effect for expression of tau protein Src from cell A7r5 induced by Ang? and phosphorylation from Tianma Gouteng Decoction.Through Tianma Gouteng Decoction and its ingredients to cell A7r5 induced by Ang II,the method of Western-Blot is taken to investigate changes of Src and phosphorylation tau protein expression in cell.The result demonstrates that Tianma Gouteng Decoction and its effective ingredients adjust circadian rhythm changes for clock gene of cell by reducing Src and expression of phosphorylation tau protein.(2)The intervening effect for protein expression of Src in cell A7r5 induced by Ang?managed by inhibitor of isorhynchophylline.Through A7r5 cell managed by inhibitor PTK adjusted by Isorhynchophylline,we can see that expression for phosphorylated protein of Src is reduced,the product of phosphorylation Src is taken as down stream point of Ang?-AT1 pass and Src protein possibly participate in point for adjusting circadian rhythm of clock gene.