Screening And Function Analysis Of Molecular Markers Of AMPK And Its Downstream Genes Related To Aerobic Ability

Objects:In this study, the molecular markers associated with long-distance running ability were screened through analyzing the distribution of AMPK and its downstream genes'polymorphisms in Han of northern China long-distance runners. The molecular markers associated with long-distance running training sensitivity were screened through analyzing the relationship between genotype and the changes of physiological indexes after long-distance running training. To clarify the function of different allele contribute to gene expression, the biological function of polymorphisms loci was investigated. It may provided experimental evidence for evaluate the relationship between molecular markers and aerobic endurance capability.Methods:1. Genetic distribution for 17 SNP and 2 STR locos of AMPK a 2,MEF2A,PFK2,TORC2,ACC-β,CPT-1βand HSL genes in long-distance runners and controls were determined based on MALDI-TOF and Genescan. The relativity between gene locus and long distance running ability was analyzed.2. Genetic distribution for 9 SNP locus of AMPKα2,CPT-1βgenes in young men were determined by MALDI-TOF. The relativity between gene locus and the changes of physiological indexes after long-distance running training were analyzed.3. The molecular biological methods such as gene recombination and cell transfection were used to research the effects of Glut4 promoter carrying different allele in rs5418 contributes to the expression activity of reporter gene.Results and conclusions:1. Polymorphisms and their association with long distance performance suggests, (1)For AMPKα2, the G allele (rs2796516) might be served as molecular marker of elite female long distance runners. AC haplotype (rs2746342-rs11206889) might be used as molecular marker of elite male International long distance runners. CC haplotype (rs2746342-rs11206889) might be used as molecular marker of elite female 5/10km long distance runners. (2)Comparing short and long chain allele split by 4, the LL genotype of (CA)n of HSLi6 might be a molecular marker of elite 5/10km long distance runners.2.Polymorphisms and their association with training sensityvity suggests, (1)for AMPK a 2, the locus with higher lever in training sensitivity include the genotype of AA(rs1418442),CC(rs11206889) and some haplotypes responded to LVMI, certain genotypes responded to IVSD and special genotypes responded to SVI/150W. (2)For CPT-1β, the genotype of AA(rs131758) had a higher lever in the training sensitivity responded to HR/RE and PWS. the haplotype of AT(rs131758-rs470117) had a higher lever in the training sensitivity responded to SVI,T and COI in rest state. Above genotypes/haplotypes might be used as molecular markers of long-distance running training sensitivity.3. is a functional molecular marker of aerobic endurance capability. The relative activity of reporter gene carrying A allele of rs5418 in promoter of Glut4 in recombinant vector is significantly higher than that of G allele. The biological activity of this loci is that different alleles can alter promoter activity significantly, thus affecting the activity of Glut4 gene expression.