The Expression Of Hypoxia Inducible Factor-1 Alpha(HIF-1?)in The Fibroblasts From Granulation Of Benign Tracheal Stenosis

Benign airway stenosis is one of the major disease,including airway occupying or scar contracture stenosis caused by tracheal tuberculosis,trauma,operation,tracheotomy,endotracheal intubation,foreign matter,benign tumor,amyloid,and airway collapsing stenosis caused by thyroid cyst and relapsing polychondriti.The etiology of benign airway stenosis is varied.With the development of technology of emergency in the intensive care unit,the morbidity rate of airway stenosis caused by the intubation and tracheotomy gradually increased.Recent studies have found that the damage of tracheal mucosa broke the local mucosal blood circulation,which leads to mucosal ischemia and hypoxia and the subsequent formation of mucosal erosion,ulcer and tracheal cartilage.Studies of animal experiments and clinical cell culture in vitro showed that hypoxia was an important factor leading to the scar formation,fibrosis and inflammation.Hypoxia-inducible factor-1(HIF-1)is a key nuclear transcriptional factor when tissue responds to hypoxia and a kind of oxygen-dependent nuclear transcription factor existed in a mammal and human cells.Under hypoxic conditions,HIF-1 could combine with hypoxia responsing elements of target genes and then activates the transcription of target genes,which included vascular endothelial growth factor(VEGF),transforming growth factor beta(TGF-?)The purpose of this study was to investigate the expression of HIF-l?its downstream target genes in the fibroblasts,which laid a foundation for the study of the formation and mechanism of benign airway stenosis in the cell level.Objective: To study the expression of HIF-1 alpha and downstreamtarget genes in the fibroblast cells from granulation tissue of patients with benign tracheal stenosis in vitro.Methods: 1 This study was designed to culture the fibroblasts in vitro through the cultivation of tissue,purify the fibroblasts by difference adherence,then digest and subculture with 0.05% Trypsin-EDTA,then identify by morphological observation,HE staining and immunocytochemical identification method.2 The observation the expression of HIF-1a and target genes in fibroblast by immunofluorescence.Results: 1 The adherent cells were typically fusiform with larger cell body,clear boundaries,strong three-dimension and high refraction under the inverted microscope.2 The results of immunohistochemistry showed that vimentin staining was positive,which a lot of brown yellow granules in the cytoplasm.3 HE staining showed that the cells were long spindle shape with clear contour and big nucleolus.4 expression of protein HIF-1 alpha and its downstream target gene of TGF beta 1 was red and vascular endothelial growth factor(VEGF)was green in cytoplasm of fibroblasts by immunofluorescence,respectively.It is suggested that HIF-1 alpha and its downstream target gene TGF beta 1 and VEGF expressed on fibroblasts.Conclusion: Our research showed that the expression of HIF-1 alpha and its downstream target gene TGF beta 1 and VEGF on the fibroblasts from human benign airway stenosis granulation tissue by the immunofluorescence method.So it laid a strong foundation for the role of HIF-1 alpha and its downstream target gene TGF beta 1 and VEGF in the formation of benign airway stenosis and mechanism in the cell level.