| Purpose:To observe the effects of klotho in myocardium and kidney tissue and the structure changes of myocardium tissue of chronic heart failure rats,curing by Shencao Tongmai Granules,Through establishing chronic heart failure(CHF)rat models.studying the machanism and target of action of Shencao Tongmai Granules on CHF by terms of Gene,protein and histomorphology,which is providing the futher method of prevention-treatment of Chinese medicine,at the same time,as the scientific basis for CHF.Material and Method:1.Through ligating anterior descending branch of coronary artery to created chronic heart failure(CHF)rat models with exhausted swim,less food and other methods,then determining whether the model of CHF rats was sucessful or not by the status and the behavior of rats,also using the Doppler echocardiography to check ejection fraction(EF)? 55% or CI ? 180ml/min/kg.2.Setting Sham operation group as control group,And the CHF models' rats randomly set as TCM,western medicine,and model group.Model group and control group did not use any medicine,only fed with distilled water which is same volume with other groups,rats in TCM group fed 9.2g medicines / Kg / d and rats in western group fed 1.5mg lisinopril / Kg / d by mice and men dose formula.4 weeks later,collecting blood from tail vein,then all rats should be killed by anesthesia,And left the heart myocardium and kidney tissue.3.Determing the concentration of BNP and klotho of heart myocardium and kidney tissue by ELISA,Western blot,and real time quantitative PCR among TCM group,Western group and control group,model group.Comparing the structure changes of histomorphology and cardiomyocytes of four groups under light microscope.Results:1.Comparing the concentration of BNP among four groups,Medicine group(western and chinese)and model grouop was significantly increased,p<0.05.The concentration of BNP of control group was 450±37.03pg/ml,model group was 1017.55±98.6pg/ml,Chinese Medicine group,Western group was 681.08±75.31pg/ml,686.53±74.86pg/ml respectively.p>0.05.2.The releasing of klotho mRNA of myocardium in model group,was significantly decreased,was 0.0004±0.00004(contrasting with the control group P <0.01);After curing by TCM and western medicine,klotho mRNA expression was significantly increased,is 0.0020±0.0009(compared with model group and control group,P <0.01,compared with the control group P <0.01);TCM group was 0.0012±0.0007(contrasting with model group,P <0.01,contrasting with the control group P <0.01).3.CHF rat model group klotho protein expression of myocardium was significantly lower,was 0.0291±0.0025(compared with control group P <0.01);by medicine intervention klotho protein of each treatment group was increased,0.2475±0.1407,(compared with model group and control group,P <0.01);TCM group was 0.2146±0.0765(contrasting with model group,P <0.01,and the control group P <0.01).4.CHF rat model group klotho mRNA of kidney tissue expression was obviously reduced,was 0.0022±0.0007(contrasting with the control group P <0.01);After curing by TCM and western medicine,the klotho mRNA was increased,Western medicine group more obvious was 0.0067±0.0020(compared with model group,P <0.01,and the control group P <0.01);Chinese medicine group 0.0063±0.0022(compared with model group,P <0.01,and the control group P <0.01).5.The releasing of klotho protein of kidney tissue was declined significantly in model group,was 0.0639±0.0203(contrasting with the control group P <0.01);After curing by TCM and western medicine,the klotho protein was increased,Western medicine group more obvious was 0.2150±0.0303(contrasting with model group,P <0.01,contrasting with the control group P <0.01);TCM group 0.2057±0.0457(contrasting with model group,P <0.01,contrasting with the control group P <0.01).6.Control group,Cardiomyocytes are intact,the muscle fibers are neat and orderly,the Z line is clear and uniform,the sarcoma has obvious periodicity,the myocardium has no edema in the myocardium,and there are a large number of mitochondria in the muscle fibers.model group,Sarcomere structure is unclear or disappear,Myofibrils are broken and dissolved,Sarcomere disorder,Myocardial tissue interstitial with obvious edema,necrotic area area is large,necrosis serious,mitochondrial morphology is different,see swelling,rupture or disappear,vacuately serious.Chinese medicine group,Sarcomere clearer,there are still local muscle fibers broken,dissolved,mitochondria arranged in the filaments between the slight edema,there are still left some mitochondrial cavitation.Western chinese medcine,Sarcomere clearer,part of the muscle fibers rupture,dissolved,mitochondria arranged between the filaments,mild swelling,and less cavitation mitochondria.Conclusion:1.Ligating anterior descending branch of coronary artery with less food and exhausted swim prove that this modeling method could be successed to create the chronic heart failure by the obvious outlook and behavioral of rats and echocardiography.2.Through 4 weeks treatment of Shencao Tongmai Granules,found the concentration of BNP was decreased by ELISA,by using real time quantitative PCR,Western blot,observed the klotho mRNA and protein of myocardial tissue in rats was increased,and The myocardium was observed by electron microscopy to show that the Shencao Tongmai granules could inhibit or reverse the effect of ventricular remodeling.3.After 4 weeks treatment of Shencao Tongmai Granules,using real time quantitative PCR,Western blot,observed the klotho mRNA and protein of kidney tissue in rats was increased,to show that Shencao Tongmai Granules could inhibit kidney damages.4.According to results we get in this experiment,we could guide a new way to heal chronic heart failure,such as Delayed aging of endothelial cells,on the terms of diet,to provide appropriate quality protein to patients with chronic heart failure to protect the kidneys and delay kidney injury,meanwhile,the specific mechanisms of kidney injury associate with CHF requires futher study by scientific research and clinical trials,to verify the correctness of the these views. |
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