| ObjectiveHypertension is one of the most common cardiovascular diseases,and myocardial hypertrophy is the most common complication.The main manifestations are compensatory hypertrophy of cardiomyocytes and proliferation of mesenchymal fibroblasts.Once the formation of hypertensive myocardial hypertrophy,it would eventually lead to arrhythmia,myocardial infarction,heart failure and sudden cardiac death and other malignant cardiovascular events.Therefore,it is of great significance to explore the mechanism of hypertension myocardial hypertrophy.At present,a large number of clinical studies have confir med that acupuncture can improve heart function and reduce cardiova scular events through multiple targets and channels.Taichong and Taixi points are the original points of liver and kidney meridians.The former could regulate the liver qi and reduce blood pressure.The latter can nourish the kidney yin,and electroacupuncture at the two points can reduce blood pressure.Therefore,in this experiment,SHR rats were used to establish the model of hypertensive myocardial hypertrophy,and Taichong and Taixi points were intervened by electroacupuncture.Through PRM and Western blot techniques,the pathogenesis of hypertensive myocardial hypertrophy and the mechanism of electroa cupuncture at Taichong and Taixi points were discussed from the protein level,providing a new theoretical basis for acupuncture treatment of hypertensive myocardial hypertrophy.Methods1.Experimental group and treatment30 SHR rats were randomly divided into three groups according to their weight:model group(SHR group),acupuncture group(EA group),non acupoint group(sham group),10 rats in each group.Ten Wistars of the same week age(WKY group)were used as the control group.WKY group and SHR group:the same grasping and fixation methods were used as sham group and EA group,but acupuncture and electroacupuncture were not used;Sham group:Acupuncturing the corresponding acupoints of rats:Taichong acupoint corresponds to the front depression of the third and fourth phalanx junction of the hindlimb,and Taixi acupoint corresponds to the front 3-5mm of the heel of rats.EA group:single side Taichong and Taixi points were selected for acupuncture.After acupuncture,two needle handles were connected to the electroacupuncture therapeuticapparatus,and electroacu punc ture stimulation was given(density wave,2Hz/15Hz,strength of 1mA,20min each time,once a day,for 8 consecutive weeks).2.Evaluation indicators(1)Blood pressure and heart ratebefore the experiment and after the first,second,third,fourth,fifth,sixth,seventh and eighth weeks of the experiment,the changes of systolic blood pressure,diastolic blood pressure and heart rate in four groups were detected.(2)Heart index,left and right kidney indexafter the eighth week of the experiment,the whole heart weight,left ventricular weight,left and right kidney weight and body weight of the rats in four groups were measured,and the whole heart index,left ventricular index,left and right kidney index were calculated.(3)Morphological observation of heart and kidneyafter the eighth week of the experiment,HE staining was used to observe the changes of heart and kidney tissue in four groups and calculate the cross-sectional diameter and area of myocardial cells.(4)Expression of cardiac differential protein:after the eighth week of theexperiment,PRM technology was used to detect the expressions of cardiac differential protein in four groups.(5)Verification of detection proteinafter the eighth week of the experiment,Western blot was used to detect the myocardial expression protein of SOD1,SOD2,HSP60,HSP70,myosin light chain and fatty acid binding in four groups.Results1.The results of blood pressure and heart rate showedthe main effects of systolic blood pressure,diastolic blood pressure and heart rate treatment time were significant(P<0.01),the main effects of grouping factors were significant(P<0.01),and the interaction between time and grouping factors was significant(P<0.01)in four groups.Further,time factor was fixed at the same level.From the 0th week to the 8th week,compared with WKY group,systolic blood pressure,diastolic blood pressure and heart rate were increased in SHR group and sham group(P<0.01);Compared with SHR group,systolic blood pressure,diastolic blood pressure and heart rate were decreased in EA group(P<0.01);Compared with sham group,systolic blood pressure,diastolic blood pressure and heart rate were decreased in EA group decreased(P<0.01).2.The results of heart index showedCompared with WKY group,the whole heart weight,the whole heart index,the left ventricular weight and the left ventricular index were increased in SHR group(P<0.01),the whole heart index and the left ventricular index were increased(P<0.01,P<0.05),there was no significant difference of the whole heart weight and the left ventricular weight in Sham group(P>0.05);Compared with SHR group,the whole heart weight,the whole heart index and the left ventricular weight were decreased in EA group(P<0.01);Compared with sham group,the whole heart weight,the whole heart index,the left ventricular weight and the left ventricular were index decreased in EA group(P<0.01,P<0.01,P<0.05,P<0.05).3.The results of renal index showed thatCompared with WKY group,the left and right renal indexes were increased in SHR group(P<0.05),the left and right renal indexes were increased in Sham group(P<0.01,P<0.05);Compared with SHR group,the left and right renal indexes were decreased in EA group(P<0.05);Compared with sham group,the left and right renal indexes were decreased in EA group(P<0.01,P<0.05).4.The results of the volume fraction of myocardial collagen and the ratio of ?/? collagen showed thatCompared with WKY group,the volume fraction of myocardial collagen and the ratio of ?/? collagen were increased significantly in SHR group(P<0.01),the volume fraction of myocardial collagen and the ratio of ?/?collagen were increased significantly in Sham group(P<0.01);Compared with SHR group,the volume fraction of myocardial collagen and the ratio of ?/?collagen were decreased significantly in EA group(P<0.01);Compared with sham group,the volume fraction of myocardial collagen and the ratio of ?/?collagen were decreased significantly in EA group(P<0.01).5.The results of the cross-sectional diameter and area of myocardial cells showed that:Compared with WKY group,the cross-sectional diameter and area of myocardial cells were increased significantly in SHR group(P<0.01),the cross-sectional diameter of myocardial cells were increased(P<0.01),there was no significant difference of area of myocardial cells in Sham group(P>0.05);Compared with SHR group,the cross-sectional diameter and area of myocardial cells were decreased in EA group(P<0.01);Compared with sham group,the cross-sectional diameter and area of myocardial were cells decreased significantly in EA group(P<0.01,P<0.05).6.The morphological observation results of heart and kidney showed that:Myocardial HE staining:SHR group:Compared with WKY group,the arrangement of myocardial fibers was disordered and broken,myocardial cells were hypertrophied,and interstitial edema was obvious,myocardial interstitial infiltration and fibrocyte proliferation were obvious;EA group:Compared with SHR group and sham group,the swelling and hypertrophy of myocardial cells were decreased,interstitial edema was reduced,and myocardial fibers were orderly arranged,myocardial interstitial inflammatory cells were soaked.Renal HE staining:SHR group:Compared with WKY group,protein particles were separated out;inflammatory cells infiltrated.EA group:Compared with SHR group and sham group,protein particles precipitation and inflammatory cell infiltration decreased.7.the results of differential protein expressions showed thatCompared with WKY group,the protein expressions of creatine kinase B,creatine kinase M,SOD2 and HSP27 were decreased(P<0.01),the protein expres sions of ATP synthetase,HSP70,myosin light chain 3,fatty acid binding protein,myosin light chain 2,SOD1,sorting connexin protein and MAPK kinase were decreased(P<0.05),the protein expressions of type ? collagen,?-enolase,propionyl-CoA-carboxylase and calpain were increased were increased(P<0.01,P<0.01,P<0.05,P<0.05),there was no significant difference in the protein expressions of HSP60 and dynamic-related protein 1(P>0.05)in SHR group,the protein expressions of creatine kinase B,creatine kinase M,myoglobin light chain 3,fatty acid binding protein were decreased(P<0.01),the protein expressions of ATP synthetase,dynamic-related protein 1,SOD1,SOD2,HSP27 were decreased(P<0.05),the protein expressions of type? collagen,propionyl-CoA-carboxylase,?-enolase,calpain were decre Mased(P<0.01,P<0.01,P<0.01,P<0.05),there was no significant difference of the protein expressions of HSP60,HSP70,myoglobin light chain 2,sorting connexin protein,MAPK kinase in Sham group(P>0.05);Compared with SHR group,the protein expressions of creatine kinase B and creatine kinase M were increased(P<0.01),the protein expressions of ATP synthetase,HSP70,myosin light chain 3,fatty acid binding protein,myosin light chain 2,sorting connexin protein,MAPK kinase were increased(P<0.05),the protein expressions of type I collagen,calpain,?-enolase and propionyl-CoA-carboxylase were decreased(P<0.01,P<0.01,P<0.01,P<0.05),There was no significant difference of the expression of HSP60,dynamic-related protein 1,SOD1,SOD2,HSP27(P>0.05)in EA group;Compared with Sham group,The protein expressions of creatine kinase B,creatine kinase M,fatty acid binding protein,dynamic-related protein 1 were increased(P<0.01);The prot ein expressions of ATP synthetase,HSP70,myoglobin light chain 3,myoglobin light chain 2 were increased(P<0.05),The protein expres sions of type ? collagen,propionyl-CoA-carboxylase,calpain,a-enola se were decreased(P<0.01),There was no significant difference in the expression of SOD1,SOD2,HSP27,sorting connexin protein,HSP60,MAPK kinase in EA group(P>0.05).8,The results of protein test showed thatCompared with WKY group,the protein expressions of SOD1,SOD2,myosin light chain,fatty acid binding protein and HSP70 was decreased(P<0.05,P<0.01,P<0.05,P<0.05,P<0.05),there was no significant difference in the protein expressions of HSP60 in SHR group(P>0.05),There was no significant difference in the protein expressions of SOD1,SOD2,HSP60,HSP70,myosin light chain and fatty acid binding in Sham group(P>0.05);Compared with SHR group,the protein expressions of SOD1,SOD2,HSP60,HSP70,myosin light chain and fatty acid binding protein were increased significantly in EA group(P<0.01,P<0.05,P<0.05,P<0.01,P<0.01,P<0.01);Compared with Sham group,the protein expressions of SOD1,myosin light chain,HSP70,fatty acid binding protein were increased(P<0.05),there was no significant difference of the protein expressions of SOD2,HSP60 in EA group(P>0.05).Conclusion1.Systolic pressure,diastolic pressure,heart rate,whole heart weight,left ventricular weight,whole heart index,left ventricular index,left and right kidney index,cross-sectional diameter and area of myocardial cells,myocardial collagen fiber volume,?/? collagen fiber ratio were all increased in SHR rats,while electroacupuncture could reverse the above mentioned indexes.2.From the change of HE staining picture:cardiomyocyte hypertrophy,interstitial edema,interstitial inflammatory cell infiltration,fibrocyte proliferation in myocardium tissue and Precipitation of protein particles,inflammatory cell infiltration in renal tissue in SHR group while electroacupuncture can restore the pathological performance of appealing myocardium.3.In SHR rats,the protein expressions of creatine kinase B,creatine kinase M,ATP synthetase,myosin light chain 3,fatty acid binding protein,myosin light chain 2,SOD1,SOD2,HSP27,HSP70,sorting connexin protein,MAPK kinase protein decreased,while the expression of type ? collagen,a-enolase,calpain,propionyl-CoA-carboxylase were increased.4.Electroacupuncture can increase the protein expressions of creatine kinase B,creatine kinase M,ATP synthetase,myosin light chain 3,fatty acid binding protein,myosin light chain 2,sorting connexin protein,HSP70,MAPK kinase protein in SHR rats,but decrease the expression of calpain,type ?collagen,?-enolase,propionyl-CoA-carboxylase protein. |
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