Effects Of Adiponectin On The Expressions Of Adipor1、adipor2 And T-cadherin In Cardiomyocytes Injure After Hypoxia-reoxygenation

PartⅠDistribution and expression of adiponectin receptors in SD ratcardiomyocytesObjective To investigate the distribution and expression of adiponectin receptors in SD ratcardiomyocytes.Methods1. Primary myocardial cells were obtained from neonatal rats by enzymatic digestionmethod.The molecular markerα-actin of the cardiomyocytes was observed byimmunocytochemistry.The morphology of neonatal rat cardiomyocytes was studied by phasecontramicroscope.2. The cells cultured for 72~96 h were used in the experiments, The expressions of AdipoR1、AdipoR2、T-cadherin were detected by RT-PCR and immunocytochemistry.Results1. Cardiomyocytes were confirmed and the purity was more than 95%.2. The mRNA and proteins of AdipoR1、AdipoR2 and T-cadherin were all observed in SD ratcardiomyocytes, while the expression of AdipoR1 and T-cadherin were much higher than theexpression of AdipoR2 in cardiomyocytes(P<0.01).Conclusions1. AdipoR1、AdipoR2 and T-cadherin were all distributed in SD rat cardiomyocytes.2. AdipoR1、AdipoR2 and T-cadherin were all distributed in SD rat cardiomyocytes, especiallyAdipoR1 and T-cadherin, while AdipoR2 expressed lower.PartⅡEffects of adiponectin on the expressions of AdipoR1、AdipoR2 andT-cadherin in cardiomyocytes injure after hypoxia-reoxygenationObjective To investigate the effects of adiponectin on the expression of AdipoR1、AdipoR2and T-cadherin on cultured SD rat cardiomyocytes by hypoxia-reoxygenation.Methods1. Primary myocardial cells were obtained from neonatal rats by enzymatic digestion method.The molecular markerα-actin of the cardiomyocytes was observed by immunocytochemistry.Themorphology of neonatal rat cardiomyocytes was studied by phase contramicroscope.2. The cells cultured for 96 h were used in the experiments, and they were divided into groupsrandomly: control group, H/R group, H/R+3 ug/ml APN group, H/R+10 ug/ml APN group,H/R+20 ug/ml APN group, H/R+30 ug/ml APN group. 3. The changes of morphology of cardiomyocytes were observed by phase contractedmicroscope.The content of LDH was detected by chemistry chromatometry; The ratescardiomycocytes apotosis was detected by flowcytometry and terminal deoxynucleotidyltransferrase-mediated dUTP nick end labeling (TUNEL).4. The expression of AdipoR1,AdipoR2,T-cadherin were detected by RT-PCR and Westernblotting.Results1. Cardiomyocytes were confirmed and the purity was more than 95%.2. Compared with control group,the rate of apoptosis in group H/R was significantly increased,the TUNEL-positive cells(green) increased significantly,the release of LDH wasincreased,Pretreatment with APN, the above changes were significantly reversed.3. Compared with control group, the expresstion of T-cad mRNA and the protein was decreasedin group H/R. Pretreatment with APN, the above changes were significantly reversed. Comparedwith group H/R,the indexes improved obviously(P<0.01) in a dose-dependent manner.4. Compared with control group, the expresstion of AdipoR1 mRNA and the protein wasdecreased in group H/R. Pretreatment with APN, the above changes were significantly reversed.Compared with group H/R,the indexes improved obviously(P<0.01), but each APN treatmentgroup do not depend on the APN concentration changes.5. Compared with control group, the expresstion of AdipoR2 mRNA and the protein was in alittle down in group H/R. Pretreatment with APN, the levels of AdipoR2 mRNA and protein wasno obvious change(P＞0.01). Compared with group H/R,there was also no obvious change(P＞0.01), and levels were independent of APN concentration changes.Conclusions1. Hypoxia-reoxygenation can induce injury in rat Cardiomyocytes.2. APN can protect cardiomyocytes from the injury induced by hypoxia-reoxygenation .3. APN may be protect cardiomyocytes from hypoxia-reoxygenation injury by upregulating theT-cadherin expression. The protection may be mainly through combined with its receptor T-cad.4. AdipoR1 and AdipoR2 may be the adiponectin secondary receptors to the protection of thecardiomyocytes, and both were independent of APN concentration changes,and AdipoR2protection against damage effect of cardiomyocytes may be relatively weak.