| Objective:To observe the therapeutic effects of the classical formula of Zhen Wu Tang on rats with hypothyroid heart disease(referred to as hypothyroid heart)and to explore its mechanism of action based on AMPK/PGC-1α signaling pathway,so as to provide a theoretical and scientific basis for its clinical application and provide effective reference for scripture development.Methods:The hypothyroidism model rats were established by continuous gavage of 0.1%PTU suspension(10 mg/kg·d)for 28 days.The general condition of rats was observed,thyroid function was measured by ELISA,and cardiac function was evaluated by M-mode echocardiography,and the successful establishment of the model of hypothyroidism was comprehensively evaluated.The rats with successful modeling were randomly divided into model group(n=10),Zhenwu decoction low-dose group(n=10),Zhenwu decoction medium-dose group(n=10),Zhenwu decoction high-dose group(n=11),Zhenwu decoction positive drug(Youmetolol)group(n=10)and normal group(n=10).The dose of Zhenwu decoction in low-dose,medium-dose and high-dose groups was 7.5 g/kg·d,15 g/kg·d and 30 g/kg·d,respectively,while that in positive group was 9μg/kg·d.The general condition of the rats in each group was observed and recorded after continuous intragastric administration for 28 days.The cardiac function of the rats in each group was detected by M-mode echocardiography,the serum concentrations of TSH,FT4,T,E2 and CRH were detected by ELISA,and the pathological changes of the heart tissue of the rats were observed by HE staining,and the efficacy of Zhenwu Decoction on hypothyroidism model rats was observed.The concentration of myocardial ATP in each group was detected by chemiluminescence method,and the protein expressions of AMPK,p-AMPK,PGC-1α,Nrfl and Tfam in heart tissue were detected by Western Blot,so as to explore the mechanism of Zhenwu Decoction in the intervention of hypothyroidism.Results:1.Compared with the normal group,body mass,anal temperature and food intake were significantly lower(P<0.01),water consumption was significantly reduced(P<0.05),urine output was significantly increased(P<0.05),TSH concentration was significantly increased(P<0.01),FT4 concentration was significantly decreased(P<0.05),LVEDD and LVEDV were significantly reduced(P<0.01),and LVESD and LVESV were significantly increased(P<0.01).LVESD and LVESV significantly increased(P<0.01),HR significantly slowed(P<0.01),and SV,CO,LVEF,and LVFS significantly decreased(P<0.01).In conclusion,the hypothyroid heart model was prepared successfully,and the modeling rate was 85%.2.Compared with the normal group,the body mass,anal temperature,feeding volume and drinking volume of rats in the model group decreased significantly(P<0.01)and urination volume increased significantly(P<0.01);compared with the model group,the body mass of each drug administration group increased significantly(P<0.05),anal temperature increased significantly(P<0.01),feeding volume and drinking volume of the low and medium dose groups of Zhenwu Tang and the positive drug group increased significantly(P<0.01,P<0.05),and urine output was significantly reduced in each drug administration group(P<0.01).3.ELISA results showed that compared with the normal group,the concentration of TSH was significantly increased(P<0.01)and the concentration of FT4,T,E2 and CRH was significantly decreased(P<0.01)in the model group;compared with the model group,the concentration of TSH was significantly decreased(P<0.01)and the concentration of FT4 was significantly increased(P<0.05)in each administration group,and there was an increase in the concentration of T in each administration group of Zhenwu Tang in rats The T concentration of rats in each administration group of Zhen Wu Tang had a tendency to increase,but there was no statistical difference(P>0.05),the T concentration of rats in the positive drug group increased significantly(P<0.05),E2 and CRH in Zhenwu decoction group(P<0.05,P<0.01).4.M-mode echocardiographic results showed that compared with the normal group,the LVESD and LVESV of rats in the model group were significantly increased(P<0.01),while HR was significantly slower(P<0.01)and CO,LVEF and LVFS were significantly decreased(P<0.01);compared with the model group,the HR of rats in each administration group was significantly faster(P<0.01),CO was significantly increased(P<0.01,P<0.05),and LVEF and LVFS were significantly increased(P<0.01).5.The results of cardiac index showed that the cardiac index of rats in the model group was significantly lower compared with the normal group(P<0.05);compared with the model group,the cardiac index of rats in each administration group tended to increase,but there was no statistical difference(P>0.05).6.HE staining results showed that the arrangement of cardiomyocytes in the model group was disordered,distorted and blurred,and nuclear fragments were commonly found in cardiomyocytes;the morphology and structure of cardiomyocytes in each drugadministered group were improved and the degree of damage was reduced.7.The results of ATP assay showed that the myocardial ATP content of rats in the model group was significantly reduced compared with the normal group(P<0.01);compared with the model group,the ATP content in the myocardium of rats in each administration group was significantly increased(P<0.01).8.Western Blot results showed that the p-AMPK/AMPK ratio,PGC-1α,Nrfl and Tfam were significantly decreased in rats in the model group compared with the normal group(P<0.05,P<0.01);compared with the model group,the p-AMPK/AMPK ratio was significantly increased in rats in the medium and high dose groups of Zhenwu Tang and the positive drug group(P<0.05).The expression of PGC-1α,Nrfl and Tfam was significantly higher in rats of each administration group(P<0.05,P<0.01).Conclusion:1.The hypothyroid heart model could be successfully replicated by continuous gavage with PTU suspension for 28 days.2.The formula of Zhen Wu Tang can significantly improve the general condition,thyroid function and cardiac function of hypothyroid heart model rats and reduce the damage of heart tissue.3.Chenwu Decoction can activate AMPK/PGC-1α signaling pathway,promote AMPK protein phosphorylation,up-regulate the expression of PGC-1α,Nrfl and Tfam,increase the content of ATP in myocardial tissue,improve mitochondrial function,alleviate myocardial energy metabolism disorders,and thus alleviate myocardial damage in rats with hypothyroidism. |
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