| AIM:Experimental study shows that the drug(Ber and Rut) can improve the spontaneously hypertensive rat heart function,and inhibition of vascular remodeling.In this study,via observe the blood pressure of SHR which taking capsules to clear the role of lower blood pressure,application the protein separation and identification techniques to the blood of SHR,analysis by 2-DE and establish spectrometry of different proteins.Via mass spectrometry of different proteins that treatment by LY caspule,discuss the Step-down mechanism of Chinese medicine to provide scientific basis for the LY caspule in the treatment of hypertension-related basic research,and discuss antihypertensive medicine target protein molecules,increase the basic and clinical study of Chinese medicine treatment of hypertension,also to provide the theory and the scientific basis for the clinical application of Chinese medicine.METHODS:12-week-old SHR and Wky rats were used in this study.30 SHR rats were divided into Model group,LY caspule group,Ogilvy & Mather cilexetil group and 10 rats were Wky group.Each of the five male and female rats.Wky group and Model group fed with NS,as LY caspule group with suspension of Ber and Rut(3:1),as Ogilvy & Mather cilexetil group with ester solution olmesartan by Ig for 4 weeks.All the rats were used the pressure method and measured the arterial blood pressure in the fourth weekend.We sampled aortic blood when the rats were anesthesia,mixed and low-temperature centrifugal(2500rmp,4℃).Then we can separate the plasma and put the serum in the poly ethylene plastic tube.All of the samples were keeped in the refrigerator at -80℃.In the proteomic analysis experiment,firstly,we measured protein content by bradford,separated protein in different PH gradient by IEF,then separated protein in different Mr gradient by SDS-PAGE and then used the technique of silver stain.After that we can get the two-dimensional gel protein map of Model group,LY caspule group,Ogilvy & Mather cilexetil group and Wky group.By imagemaster computational software we got the differential expressed proteins of the samples.We used MALDI-TOF-MS to determine the PMF of the differential expressed proteins,searched and determined the protein that the peptide corresponding to,then got the information of the differential expressed proteins.RESULT:1.LY caspule on blood pressure:Compared with the Model group,the rats' blood pressure of LY caspule group,Ogilvy & Mather cilexetil group were significantly reduced(P＜0.01).2.Successful construction of four 2DE map of proteins.By analyzing the difference of four 2DE map of proteins,there were 11 significant different protein spots. Compared with the Wky group,4 protein spots were detected up-regulately and 6 spots down-regulately in Model group.3.After the the treatment of LY capsule and Ogilvy & Mather,the down-regulately, and the up-regulately were closed to the Wky group.In the LY capsule group,there was a new protein spots.4.Differences in protein spots 11 through the enzyme-gel digestion,mass spectrometry and database search identified a total of six protein spots,and five protein spots have not been satisfactory due to the peptide mass fingerprint database or search can not be the result of higher reliability can not be identified.Identification of protein are:E3 ubiquitin protein ligase,inner membrane protein mitochondrial,zinc finger protein,Rho GTPase activating protein,T cell receptor,isoform CRA_a.Function of these proteins involved in cell metabolism,signal transduction, electron transport,such as the cytoskeleton.CONCLUSION:1.The effect to hypertension of LY caspule is existing.LY caspule can lower blood pressure of SHR Lasting and stable.2.By analyzing the difference of four 2DE map of proteins,there were 10 significant different protein spots in Model group,there are seven of them decreased,and three of them increased.The blood protein expression of SHR which after the treatment of LY caspule had significant differences,and there was a new protein spots.3.We used MALDI-TOF-MS to determine the PMF of the 11 differential expressed proteins to search and determin the protein what the peptide corresponding to,then we can get the information of the 6 differential expressed proteins.4.After the the treatment of Ogilvy & Mather,the up-regulately Rho GTPase and the down-regulately,such as inner membrane protein mitochondrial,zinc finger protein, T cell receptor,isoform CRA were closed to the Wky group.In addition,after the the treatment of LY capsule,E3 ubiquitin protein ligase was found.5.At the same time,we observed the effect of the different protein expression which were after the the treatment of LY caspule,and discussed the mechanism of LY caspule may invovle multiple procedures or multiple factors in protein level.LY caspule is a full of great research value and application prospects Chinese Medicine. |
PDF Full Text Request