Association Study Between Adrenocortical Hormone Synthase Gene’s Polymorphism And HAPE And The Effect Of Hypoxia On The Expression Of Glucocorticoid Receptor And Inflammatory Factors

Objective To explore the relationship between adrenocortical hormone synthase gene’s polymorphism and HAPE and the effect of hypoxia on the expression of glucocorticoid receptor and inflammatory factors.Methods 1.The Tag SNPs of CYP17,HSD3B1,CYP21,CYP11B1 genes in169 HAPE patients（HAPE-p）and 309 matched healthy controls（HAPE-r）were genotyped by Sequenom Mass ARRAY?SNP,and the genotype,allele frequency,odds ratio（ORs）and 95%were calculated.2.The human macrophages（HAM）,alveolar epithelial cells（HPAEpi C）and pulmonary artery endothelial cells（HPAEC）were cultured under normal oxygen（21%O₂）and low oxygen（3%O₂）by 6 hr,12 hr and 24 hr.The expression of proteins and m RNA of HIF-1α,HIF-2α,GRα,NF-k B,TNF-α,IL-6 were detected by Elisa and RT-q PCR in the culture supernatant and cells.3.78 SD rats were randomly divided into normoxic control group（n=6）,LPS group（n=24）,hypoxia experimental group（n=24）and hypoxia+LPS group（n=24）.The three experimental groups were divided into four time points of 6 hr,12 hr,24 hr and 36 hr,with 6 rats in each group.Hypobaric oxygen chamber simulates an altitude of 5000 m,and lipopolysaccharide（LPS）induces basic inflammation（0.5 mg/kg by tail vein injection）.The normoxic control group and LPS group are outside the hypobaric oxygen chamber,while the hypoxia group and hypoxia+LPS group are placed in the hypobaric oxygen chamber.The lung tissue were colleted at 6 hr,12 hr,24 hr and 36 hr after tail vein injection of LPS,the morphology of lung tissue was observed by HE staining,and the gene expression of HIF-1α,HIF-2α,GRα,NF-κB,TGF-β,IL-10,VEGF were detected by RT-q PCR.Results 1.Four SNPs,including the allele C of rs6203（p=0.034,OR[95%CI]=1.344[1.022-1.767]）in HSD3B1,allele G of rs3740397（p=0.044,OR[95%CI]=1.314[1.007-1.714]）and allele C of rs10786712（p=0.039,OR[95%CI]=0.751[0.572-0.986]）in CYP17A1,and allele T of rs6402（p=0.006,OR[95%CI]=0.504[0.306-0.830]）in CYP11B1,were significantly associated with HAPE.The distribution of the genotypes of these SNPs also significantly differed between the HAPE-p and HAPE-r groups.Moreover,rs10883783,rs4919686,rs3740397,rs3824755,and rs10786712 were present in a linkage disequilibrium（LD）block within CYP17A1.Haplotype analysis revealed that six haplotypes within that LD block differed significantly between the two groups（p<0.05）.The haplotypes TACCT（p=0.049,OR[95%CI]=1.536[1.000-2.359]）and TACGT（p=0.000,OR[95%CI]=12.706[5.078-31.795]）were associated with HAPE risk.In contrast,the haplotypes AAGGC（p=0.038,OR[95%CI]=0.492[0.249-0.973]）,TACCC（p=0.039,OR[95%CI]=0.551[0.311-0.976]）,TACGC（p=0.000,OR[95%CI]=0.014[0.001-0.130]）,and TAGGT（p=0.025,OR[95%CI]=0.720[0.541-0.960]）were associated with a lower risk of HAPE.2.The protein and m RNA expression level of HIF-1α,HIF-2α,NF-k B,TNF-α,IL-6 in HAM,HPAEpi C and HPAEC cells were upregulated under hypoxic condition comparing with that in normoxic condition（p<0.05）,while the GRαwere down-regulated（p<0.05）.3.The expression of HIF-1α,HIF-2α,NF-κB,VEGF m RNA in rat lung tissue were significantly upregulated in hypoxia group and hypoxia+LPS group compared with in the normoxic control group and LPS group（p<0.05）,while the GRα,TGF-β,IL-10 m RNA were significance downregulated（p<0.05）.Conclusions 1.The 4 SNPs in HSD3B1（rs6203）,CYP17A1（rs3740397 and rs10786712）,CYP11B1（rs6402）genes and 6 haplotypes of CYP17A1 gene correlated with HAPE,the potential molecular mechanisms need further study.2.The expression of proteins and m RNA of the HIF-1α,HIF-2α,NF-k B,TNF-α,IL-6 were upregulated in human macrophages,alveolar epithelial cells and pulmonary artery endothelial cells under hypoxia,while the GRαwere downregulated,and the relationship between those genes needs to be further explored.3.The expression of HIF-1α,HIF-2α,NF-κB,IL-10,TGF-β,VEGF m RNA in lung tissue of SD rats under LPS induced basic inflammation plus hypoxia were significantly upregulated,while the GRα,TGF-β,IL-10 were downregulated,lead to the result of inflammation imbalance in lung tissue,and the relationship between those genes needs to be further explored.