To Investigate The Mechanism Of Jiawei Shengmai Buxin Dan In The Treatment Of Diabetic Cardiomyopathy With Qi Deficiency And Phlegm Obstruction Syndrome Based On Adiponectin-mediated Energy Metabolism Pathway

Diabetic cardiomyopathy(DCM)is one of microvascular complications in diabetes,which is characterizedwith high incidence,high motality and occult.It is classified into xiaoke disease,heart palpitations inChinese medicine.The disease location of DCM is heart,which is secondary to diabetes,wih deficiency of both qi and yin,phlegm obstruction and blood stasis for the onset.Jiawei Shengmai Buxin Dan(BXD)has been used more than 10 yearsin the first affiliated hospital of hunan university of Chinese medicine,with good effect in heart disease,which was proved in experiments before that can improve animal's cardiac function,improve insulin resistance.However,the modern biological basis of deficiency in origin,phlegm and blood stasis pathogenesis of DCM is still lacking,the mechanism of BXD in the treatment of DCM has not been elucidated.Adiponectin(APN)is considered an essence of“Qi”,in Chinese medicine,with definite effect of protecting cardiac muscle.To this aim,this project is carried out as following:Aim1.The relationship between APN and DCM characterized by Qi deficiency and phlegm obstruction;2.The biological basis of DCM in Chinese medicine based on adiponectin;3.Mechanism of BXD on DCM.Methods1.Using general characteristic,behavioral tests,energy metabolism,ultrasonic cardiogram,Elisa and so on,to identifyif db/db mice be suitable as DCM model characterized by Qi deficiency and phlegm obstruction;and to investigate the relationship between APN and pathogenesis of DCM.2.Elisa was employed to investigate the changes of adiponectin and insulin,Using AAV mediated APN cDNA to restore the deficiency of APN,then illustrated the role of APN in the pathogenesis of DCM in Chinese medicine.3.BXD was given to db/db mice from the age of 16 weeks old for 8 weeks continuous,then,the effect was investigated by behavioral tests,western blot and so on.Using siRNA interference technology,to elucidate the role of APN in treatment of DCM induced by BXD.Results1.General characteristic investigation of db/db mice showed that there is a significantly obesity,high blood glucose level and insulin resistance,indicated that the diabetes in db/db mice belonged to type 2 diabetes.Behavioral tests showed that there are significantly Qi deficiency symptoms characters without age dependence from 8 weeks old.However,the energy metabolism in the heart changed significantly in db/db mice with age dependence,indicated that there is a remarkable Qi deficiency in the heart.Meanwhile,the levels of APN showed similar changes with Qi,as manifested by the loss of age dependence in plasma,and continuous decline with ages in the heart,suggested that APN maybe one of biological basis of Qi in Chinese medicine.2.AAV-APN infection restored the symptoms of deficiency of Qi in db/db mice,as exhibited by the improved metabolic flexibility,repressed uptake and oxidation of fatty acid,increased glucose utility,meanwhile,the lipidtxicity was attenuated as evidenced by decreased lipid accumulation and its metabolites,DAG and ceramide.All of these results suggested that APN improved the metabolic flexibility in DCM.AMPK and PPARs are major modulators for the energy metabolism in the heart,both of them were invovled into fatty acid and glucose metabolism.As the key transcriptor,PPAR-? mediated many gene expression,including fatty acid and glucose uptake,oxidations,and so on.In the present study,we found there is a significantly decrease of AMPK activity,increase of PPAR-?.Consequently,the genes expression which modulated by PPAR-? were also upregulated significantly.Supplementary of APN restored them significantly.More importantly,APN repressed the glycoen deposition and fibrosis significantly in DCM.3.BXD improved DCM significantly as manifested by:1)Administration of BXD improved metabolic flexility significantly,restored the ratio between uptake and oxidation of fatty acid,repressed lipidtoxicity,increased glucose utility;2)BXD attenuated cardiac function of pump,but no effect on ventricular dilatation;3)APN invovled into the improvement of BXD on DCM.Conclusions1.db/db mice showed significantly pathogenesis in TCM,which characterized by Qi deficiency and phlegm obstruction.2.APN is not only the biological basis of Qi in general,but also as the basis of Qi in the heart.3.BXD could improve DCM,and APN was invovled into this processing.