| Objective: Pulmonary fibrosis(PF)is a lung disease,characterized by the transition of epithelial cells to mesenchymal cells of the lung and parenchymal inflammation.Its pathogenesis includes many complicated factors.The damage of pulmonary epithelial cells,especially alveolar type Ⅱ epithelial cells(ATⅡ),is an essential prerequisite for the occurrence and development of PF.When the lung tissue is damaged,the tissue around and between the alveoli is thickened and scarred,causing lung ventilation disorder.As the PF worsen,the lung tissue gradually loses its normal function.In lung tissue,epithelial sodium channels(ENa C)mainly express in ATⅡ and they are the rate-limiting step in alveolar fluid clearance.In addition,studies have shown that bone marrow mesenchymal stem cell conditioned media(BMSC-CM)may have a good effect on the treatment of pulmonary fibrosis,and ferulic acid(FA)also shows a good advantage in the treatment of liver and kidney fibrosis.We deduce that ENa C may be associated with the formation of PF,and ENa C expression in PF tissue will be significantly different from normal lung tissue.This study was designed to investigate the relationship between ATⅡ-quantityrelated ENa C and PF,to clarify the expression of ENa C in the PF model,to explore possible mechanisms,and to discover the possibility of FA involved in curing PF through regulating ENa C.Methods: 1.Bleomycin was applied on mice to establish PF model.2.Detection of changes in body weight and survival rate during the modelling period.3.The distribution of normal lung tissue and PF tissue was observed by Masson staining.4.The detection of α-smooth muscle actin(α-SMA)and α-and γ-ENa C expression was achieved by Western blot at the protein level of PF tissue model.5.Real-time polymerase chain reaction(q RTPCR)was applied to measure the expression of transforming growth factor-β(TGF-β),α-SMA and α-,β-and γ-ENa C at the m RNA level of PF tissue.6.CCK-8 cell viability assay was performed to detect the effects of BMSC-CM and FA on the survival rate of ATⅡ in PF cellular model caused by TGF-β.7.Western blot was applied in the measurement of PF caused by TGF-β to determine the effect of BMSC-CM and FA on α-and γ-ENa C,α-SMA,Epithelial-cadherin(E-cadherin)and fibronectin(FN)of ATⅡ at the protein level.8.The effect of FA on m RNA expression of these markers above in PF model induced by TGF-β1 was measured by Real-time polymerase chain reaction.Results: 1.Masson staining showed that the histopathological morphology of lung tissue in mice was significantly changed after modelling by Belomycin,and the characteristics of PF were obvious.2.The body weight of mice treated with bleomycin experienced a trend of decreasing first and then increasing.3.With the increase of modelling time,the survival rate of mice showed a decreased trend.4.The expression of α-SMA and TGF-β in mouse lung tissue with PF increased at the protein and transcriptional levels after modelling,but that of α-and β-ENa C showed the opposite trend.5.BMSC-CM increased the expression of ENa C and Pf-related marker E-cadherin but decreased the expression of α-SMA and FN at protein level.6.After treated with FA,the expression of ATⅡ-quantityrelated ENa C increased and the expression of PF-related markers,E-cadherin increased but FN decreased in protein and m RNA level.Conclusion: In fibrosis,the expression of ATⅡ-quantity-related ENa C in the lung tissue and primary cultured ATⅡ decreased.Moreover,FA and BMSC-CM may involve in the process of epithelial to mesenchymal by increasing the expression of ENa C. |
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