Effects Of Core Fucosylation On Pulmonary Interstitial Fibrosis And EMT

Idiopathic Pulmonary Fibrosis(IPF)is an irreversible chronic disease that eventually progresses to ILD in interstitial lung disease in respiratory failure.There is no definitive cause,and the approximate lifetime is 2.5-3.5 years Mean survival in lung cancer,and the current data show that the annual incidence rate showed an annual increase,but because of its early clinical signs and symptoms of occult diagnosis and treatment of barriers to set up according to its 2016 Chinese experts in the treatment of pulmonary fibrosis showed that the consensus,In addition to lung transplantation,there is no effective target therapy drugs to explore the mechanism of pulmonary interstitial fibrosis has become an urgent problem to be solved at this stage.After biological information has been copied,transcribed and translated to form proteins,proteins are covalently bound to each other.This process is called protein post-translational modification.With the further study of genomics,post-translational modification of proteins has been extensively studied.Glycosylation,Phosphorylation and Acetylation Because 80% of the proteins are found to be glycoproteins,glycosylation is catalyzed by a glycosyltransferase that links the sugar chain to a specific site on the protein,Affecting the physical properties of certain biological macromolecules,regulating signal transduction,and receptor activation in the pathway,thereby participating in many pathological and physiological processes.1.6 Core fucosylation(FUT8)is a method that combines guanosine diphosphate fucose GDP-FUC)transferred to the innermost N-acetylglucosamine 6 carbon atoms forming glycosidic bonds,is the only key enzyme in mammals,less expressed in normal tissues,the previous study found that in the pathological changes of renal fibrosis The expression is increased during the course of the process,so we hypothesized whether or not it participates in the process of pulmonary interstitial fibrosis.Previous studies have found that pulmonary interstitial fibrosis is mainly the formation of large numbers of myofibroblasts and large deposits of extracellular matrix,alveolar structure loss,resulting in irreversible damage,so the source of myofibroblasts become blocked interstitial lung fibrosis In recent years,epithelial-mesenchymal transition(EMT)has become a hotspot in the mechanism research.EMT is an epithelial cell gradually transformed into myofibroblasts.Α-SMA and E-caderin are considered as epithelial-mesenchymal transitional Marker,the expression of α-SMA is a characteristic of myofibroblasts,responsible for tissue remodeling and fibrosis,E-caderin is a characteristic of lung epithelial cells,its fibrosis gradually converted to N-cadherin,EMT is more Gene,multi-signal pathways [17]-[20],in the past drug research,most of the blocking single pathway to inhibit EMT process,the effect is not good,according to the literature,EMT classic pathway TGF,EGFR,WNT and other multi-pathway Of the key proteins are glycoproteins,and modified by FUT8,whether we can interfere with the EMT process from the perspective of inhibiting FUT8 affect the occurrence and development of interstitial fibrosis.Part I Observes the Effect of Core Fucosylation on Pulmonary FibrosisPurpose: Suppression of core fucosylation can affect bleomycin-induced pulmonary interstitial fibrosis.Methods: The experiment was divided into Control group,FUT8 sh RNA group,BLM group and BLM + FUT8 sh RNA group.The model of pulmonary fibrosis was established by intraperitoneal injection of bleomycin in mice,and the core fucosylation.Whether its expression in its model,the determination of adenovirus titer,injection of adenovirus into the tail vein of mice to make it silence FUT8 gene,and its expression level was detected by western.The gross pathological changes of the four groups were observed with HE and MASSON,and pathological analysis was performed.The expression of extracellular matrix(collagen I and collagen III)was detected by immunohistochemistry and western bolt.Results: Immunofluorescence showed that the expression of LCA was significantly increased in BLM group.Masson and HE staining was used to observe the decrease of alveolar interstitial cells,thinning of pulmonary interstitium and pulmonary fibrosis in FUT8 sh RNA + BLM group compared with BLM group.Immunohistochemistry And immunoprecipitation Collagen Ⅰand Collagen Ⅲin FUT8 sh RNA + BLM group compared with BLM group decreased significantly.Conclusion:Core fucosylation exists in pulmonary interstitial fibrosis.Pulmonary interstitial fibrosis.Part II Observe the Effect of Core Fucosylation on EMTPurpose: Core fucosylation can affect EMT in pulmonary interstitial fibrosis.Methods: Control group,FUT8 sh RNA group,BLM group and BLM + FUT8 sh RNA group were used to detect the expressions of α-SMA and E-caderin in the four groups by western and immunofluorescence.The expressions of α-SMA and LCA,E-caderin Immunofluorescence with LCA to see if core fucosylation can affect the EMT process.Results: The expression of E-cadherin in BLM+ FUT8 sh RNA group was weaker than that in fibrosis group,but the expression of a-SMA was also decreased Weakened.As the expression of LCA decreased,the expression of E-cadrein increased.Conclusion:Inhibition of core fucosylation can block EMT in pulmonary interstitial fibrosisn be inhibited by inhibiting core fucosylation.