The Role Of Lxrs Agonist T0901317 On The Expression Of Fat/cd36 Mrna In The Adult And Sd Rat Skeletal Muscle Cells And Fat/cd36 Gene Polymorphism With Type 2 Diabetes

Objective To establish the methods for purification,culture, identification and biological characteristics of adult human and rat skeletal muscle satellite cells(SCs).Methods Human skeletal muscle cell and SD rat skeletal muscle cell were obtained by the two-steps method of collegenase-1 and trypsin,and were subjected to primary as well as secondary culture in vitro.Morphological characteristics,myotube formation and growth curve were observed to evaluate the proliferative and differentiation ability of SCs.The SCs were identified with cellular immuno-chemical stain.Results The two-steps method with collegenase-l and trypsin was reliable for collection of SCs.The cells showed high proliferative ability in the proliferative media and could form myotubes in differentiation media.The satellite cells of adult rat and human can be proliferated and differentiated in different serum concent ration,and can be identified by desmin stain.The growth periods in growth curve of rat was shorter than that of aldult human.Conclusion The SCs cultured in vitro from rat skeletal muscle or human skeletal muscle have high proliferative and differentiation ability and maintain their biological characteristics.They can be used to the continual study. Objective To investigate the Liver X receptors agonists T0901317's effection on expression of FAT/CD36 gene mRNA in aldult human skeletal muscle cell.Methods Myotubes from humans were exposed to different T0901317 concentrations(0,0.5,and 1.0μmol/l)for 24 hours before experiments were performed.Then the expression of FAT/CD36 mRNA in skeletal muscle cell of each experimental human group was detected by SYBR Green I real-time quantitative polymerase chain reaction.The relative data were compared among groups by 2^{-△△Ct}method.Results(1)The Ct mean of control group,T0901317(0.5μmol/l)group,T0901317(1μmol/l)group was analyzed and the difference has significance(P＜0.05).(2)The expression of FAT/CD36 mRNA with Liver X receptors agonists T0901317 in human skeletal muscle cell in the T0901317(0.5μmol/l)group and T0901317(1μmol/l)group was 2.91 times and 3.03 times than the control group.Conclusion The expression of FAT/CD36 mRNA in human skeletal muscle cell afer the treatment of Liver X receptors agonists T0901317 was increasing and the fatty acid in human skeletal muscle cell increased,so we may propose that T0901317 may increase the risk of resistance in aldult human skeletal muscle. Objective To investigate the Liver X receptors agonists T0901317's effection on expression of FAT/CD36 gene mRNA in SD rat skeletal muscle cell.Metbods Myotubes from SD rats were exposed to different T0901317 concentrations(0,0.5,and 1.0μmol/l)for 24 hours before experiments were performed.Then the expression of FAT/CD36 gene mRNA in skeletal muscle cell of each experimental human group was detected by SYBR Green I real-time quantitative polymerase chain reaction.The relative data were compared among groups by 2^{-△△Ct}method.Results The Ct mean of control group,T0901317(0.5βmol/l)group,T0901317(1μmol/l)group was analyzed and the difference has no significance in the three groups.Conclusion The expression of FAT/CD36 gene in SD rat skeletal muscle cell after the treatment of Liver X receptors agonists T0901317 was the same as control group,so we propose that T0901317 may not increase the risk of skeletal muscle resistance of SD rats. Objective To study the correlation between fatty acid translocase (FAT/CD36)gene promoter region -3489C/T and code region 478C/T polymorphism and type 2 diabetes mellitus.Methods -3489C/T and 478C/T polymorphism were screened in 196 type 2 diabetic patients and 119 controls by polymerase chain reaction restriction fragment lenth polymorphism method(PCR-RFLP).Results(1)All subjects proved to be homozygote wildtypes(CC)for the 478C→T substitution.(2)No significant differences were found in alleles and genotype frequencies of FAT/CD36 gene -3489C/T polymorphism between diabetic patients and non-diabetic contronl.Conclusion FAT/CD36 gene -3489C/T or 478C/T polymprphism was not significantly associated with type 2 diabetes mellitus in Han people of Yunnan.