中性粒细胞条件性敲除Hif-1α基因小鼠的构建和鉴定OA

Aim To construct neutrophil-specific conditional hypoxia-inducible factor 1α(HIF-1α)knockout mice and ana-lyze their genotypes,providing an animal model for studying the function of HIF-1α in neutrophils.Methods Hif-1αflox/+mice were crossed with Ly6g-iCre mice to obtain Hif-1αflox/+Ly6g-iCre+mice,which were then intercrossed to generate Hif-1αflox/floxLy6g-iCre+target mice.Genomic DNA was extracted,and genotypes were identified by PCR amplification followed by agarose gel electrophoresis.H&E staining was used to evaluate the effects of neutrophil-specific knockout on major organs including the heart,liver,spleen,lungs,and kidneys.Western blot was per-formed to detect HIF-1α protein expression in bone marrow neu-trophils and peripheral blood neutrophils under hypoxic condi-tions.Results Genotyping results confirmed the successful construction of Hif-1αflox/floxLy6g-iCre+mice.Conditional dele-tion of HIF-1α in neutrophils did not affect mouse growth,de-velopment,or reproduction.Western blot results showed that HIF-1α expression was significantly reduced in both bone mar-row and peripheral neutrophils of Hif-1αflox/floxLy6g-iCre+mice under hypoxic conditions.No obvious pathological changes were observed in major tissues and organs including the heart,liver,spleen,lungs,and kidneys.Conclusion Neutrophil-specific conditional HIF-1α knockout mice are successfully con-structed and identified,providing an experimental animal model for further investigation of the role of HIF-1α in neutrophil-related diseases.