Effects Of Regulating Hypoxia Inducible Factor-1α On The Expression Of Cytokines In Cultured Human Nasal Epithelial Cells

Objective To investigate the infects of silencing hypoxia inducible factor-1α(HIF-1α) by transfecting short hairpin RNA(shRNA) into cultured human nasal epithelial cells (HNEC) and its further regulations on the expression of vascular endothelial growth factor(VEGF)、transforming growth factor(TGF)β1 and basic fibroblast growth factor/fibroblast growth factor-2 (bFGF/FGF-2).Methods the HNEC were isolated from 10 inferior turbinate-cutting patients. The GFP marked shRNA(ad-HIF shRNA and ad-HIF shRNA-neg) were designed previously.Cells were transfected with GFP marketed adenovirus which carry the shRNA targeting HIF-la and ad-HIF shRNA-neg(scramble control).We separated cells into four groups:Blank、ad-HIF shRNA-neg transfecting、ad-HIF shRNA transfecting、Hypoxic culture.Detecting the protein and mRNA level by Western Blotting and RT-PCR.Results Compared with the Blank, Transfecting ad-HIF shRNA-neg dose not significantly influence HNEC’s HIF-1α、VEGF、TGF-β1、bFGF protein(q1=1.3167; q2=0.7513; q3=0.3251; q4=0.6592) and mRNA(ql=2.3514 q2=37.0480; q3=17.1131; q4=1.0917) expression (p>0.05).By transfecting ad-HIF shRNA, the protein and mRNA expression of HIF-la were significantly reduced 40%(q=31.4690) and 39%(q=16.5900),following,the protein and mRNA expression levels of VEGF、TGF-β1、 bFGF were significantly reduced (protein:q1=19.1571; q2=33.8090; q3=45.8940) (mRNA:ql=22.3920; q2=17.8800; q3=11.5991) (p<0.05).HNEC in Hypoxic culture, the protein and mRNA expression of HIF-la were significantly up-regulated 50%(q=32.3540) and 46% (q=15.4570), following, the protein and mRNA expression levels of VEGF、TGF-β1、bFGF were significantly up-regulated (protein:ql=20.2410; q2=35.2010;q3=47.3150) (mRNA:ql=21.4713; q2=18.3710; q3=10.3651)(p<0.05).Conclusion Silencing the expression of HIF-la by transfecting ad-HIF shRNA or up-regulating the expression of HIF-la through Hypoxic culture, can effectively inhibiting or enhance the expression of VEGF、TGF-β1、bFGF,which suggests HIF-la regulates the expression of VEGF、bFGF、TGF-β1 through direct or indirect ways.