The Mechanism Study Of Acupuncture Point Lower SHR Blood Pressure On Needle Liver And Kidney Original Research

ObjectiveStudy on effect of acupuncture acupoints by observing too Creek,Taichong,too Creek with Taichong andnon Acupoint Model of primary hypertensive rats blood pressure stimulus specific effects,quantitative proteomics and expression and effect of the medulla through the detection of the corresponding brain regions of TNF-alpha,IL-10 protein and the protein was verified the research,a preliminary study the mechanism of the antihypertensive effect of acupuncture.MethodsThe SPF class male 11W SHR 90 only,male normotensive Wistar rats were randomly divided intoacupuncture at Taichong acupuncture group,acupuncture Taixi group,non acupoint group,acupuncture at Taichong + Taixi group,model group,15 rats in each group.Normal blood pressure Wistar rats as control group.In conscious rats,downlink acupuncture twirling manipulation,acupuncture 1 times a day,the first acupuncture acupoint acupuncture 2.5min left,then right point 2.5min,each lasting for a total time of 5 min,7 days of continuous acupuncture,twist rate of 80+5/min,range 180-5 degree twist.In thedaily morning 10:00-12:00 received acupuncture treatment.Normotensive Wistar rats in blank control group,model control group rats by the method of grasping the fixed 5min,but not by acupuncture.In acupuncture after 15 min of rats were detected before and after treatment the blood pressure,measured 1 times,each rat of continuous measurement of 3-5 times,taking the average value of blood pressure.Theexperiment results after the application of immunohistochemical determination of TNF-alpha,IL-10 protein expression of the corresponding brain regions and the use of quantitative iTRAQ technology and nano upgrading two dimensional chromatography and high resolution mass spectrometry(2D-nano LC-ESI-OrbiTrap High Resolution MS/MS)isolation and identification of differentially expressed proteins in each ratmedulla oblongata.All data were statistically analyzed using SPSS16.Conclusion1 after acupuncture treatment,their blood pressure than the rats in the model group,the blood pressuredid not change significantly,no significant difference(P>0.05).Acupuncture Taichong acupoint Taixigroup,acupuncture group,acupuncture Chong Xi acupoint group rats blood pressure decreased,the difference was statistically significant(P<0.05).Acupuncture before and after treatment of blood pressureof non acupoint group rats blood pressure did not change signif icantly,no statistical significance(P>0.05).Compared between the two two groups:acupuncture group and non acupoint rats,acupunctureTaichong acupoint Taixi group,acupuncture group,acupuncture at Taichong + Taixi group rats blood pressure is reduced,the difference was statistically significant(P<0.05).With the SHR model group than in non acupoint,acupuncture group rats blood pressure did not change significantly,the difference was not statistically significant(P>0.05).2 after acupuncture treatment,the expression of the model group,Taixi group and non acupoint group rat medullary TNF-alpha,IL-10 protein decreased,Taixi group decreased the most obvious(compared with the model group,P<0.01),non acupoint group also have a certain degree of decline,but compared with the Taixi group,statistically significant differences(P<0.05);the model group,the expression of Taixigroup and non acupoint group rat striatal TNF-alpha,IL-10 protein showed an upward trend,Taixi groupelevated level is most obvious,compared with the model group,after statistics processing has a verysignificant difference(P<0.01),non acupoint group also rose to a certain extent,but compared with theTaixi group,statistically significant differences(P<0.05).Proteins identified 3 quantitative expression in the medulla of rats results.(1)compared with the normal group,model group rats medullary upregulation or downregulation of protein 19,among them the WesternBlot analysis of 6 important differences in protein,in addition to the 1 protein ATP synthase mitochondrialFO family G subtypes(ATP5L,ATP synthase mitochondrial F0 complex subunit G)can not be successfully--miscellaneous synthetic clear strip band,5 successfully completed the Western Blot analysis of proteinand quantitative proteomic results consistent.The 5 differential protein contains amino oxidase(MAO-A A,Monoamine oxidase A D(ATP6V1D),ATP enzyme,V-type proton ATPase subunit D),two hydrogen lipoic dehydrogenase(DLD,Dihydrolipoyl dehydrogenase),ATP proton ATPase 116 kDa subunit enzyme V-typea isoform 1(ATP6VOA1)and adenylate kinase 1(Adenylate kinase isoenzyme AKl).These differentially expressed proteins,mainly related to the disturb-ance of energy metabolism pathway in the medulla oblongata,including mitochondrial pyruvate dehydrogenase enzyme family,the family and the respiratory chain and other biological process.(2)compared with the model group,acupuncture at Taichong + Taixigroup SHR rat medullary up-regulated or down regulated protein 4.On one protein were Western Blotprotein was verified,verification results with the results of quantitative proteomic favorable.3.Differences in quantitative protein identified expression in the medulla of rats results.(1)compared with the normal group,model group rats medullary upregulation or downregulation of protein 19,among themthe Western Blot analysis of 6 important differences in protein,in addition to the 1 protein ATP synthasemitochondrial FO family G subtypes(ATP5L,ATP synthase mitochondrial FO complex subunit G)can not be successfully miscellaneous synthetic clear strip band,5 successfully completed the Western Blot analysis of protein and quantitative proteomic results consistent.The 5 differential protein contains ami nooxidase A(MAO-A,Monoamine oxidase A),ATP enzyme D(ATP6V1D,V-type proton ATPase subunit D),two hydrogen lipoic dehydrogenase(DLD,Dihydrolipoyl dehydrogenase),ATP proton ATPase 116 kDasubunit enzyme V-type a isoform 1(ATP6VOA1)and adenylate kinase 1(Adenylate kinase isoenzymeAKl).These differentially expressed proteins,mainly related to the disturbance of energy metabolism pathway in the medulla oblongata,including mitochondrial pyruvate dehydrogenase enzyme family,the family and the respiratory chain and other biological process.(2)compared with the model group,acupuncture at Taichong + Taixi group SHR rat medullary up-regulated or down regulated protein 4.Onone protein were Western Blot protein was verified,verification results with the results of quantitativeproteomic favorable.Results1 acupuncture Taichong,Taixi point and Taichong + Taixi point can reduce blood pressure and non acupoint,acupuncture has no antihypertensive effect.To verify the effect of acupuncture acupoint specificity,while Chong Xi acupoint in rats blood pressure lowering the highest degree,prompt group holeeffect than single point well,more close to clinical practice.2 acupuncture can regulate the expression of TNF-alpha and IL-10 in spontaneously hypertensive ratmodel of brain tissue protein.Closely related to the occurrence and development of medullary tissue in 3 hypertensive rats proteinspectrum changes and hypertension,especially the changes of energy metabolism in the medullary tissuerelated proteins have an important impact in hypertension.Acupuncture Taichong + Taixi can adjust thedifferentially expressed proteins in adult SHR medulla,which may be one of the mechanisms of acupuncture buck.