| Objective: pulmonary arterial hypertension(PAH)is a clinical syndrome characterized by progressive increase of pulmonary vascular resistance,and its main mechanism is still unclear.At present,most studies believe that pulmonary vascular remodeling is one of the most important factors leading to pulmonary hypertension.The abnormal activation of pulmonary adventitial fibroblasts(PAFs),including proliferation,migration,increased collagen secretion and phenotypic transformation,plays an important role in vascular remodeling.The expression of galectin-3(Gal-3)was increased in PAH patients and animal models.This study will explore the role of galectin-3 regulating lncrna comp induced pulmonary artery fibroblast activation in pulmonary hypertension.A new mechanism of PAFs activation was proposed to provide a new target for the treatment of PAH.Methods:(1)The serum Gal-3 level of PAH patients was measured by ELISA and compared with healthy people.Compare the correlation of Gal-3 and other pulmonary hypertension related test results.The concentration of Gal-3 in lung tissue of control rats and PAH rats modeled by monocrotaline(MCT)was detected by Western blot.Primary cultured PAFs were treated with transforming growth factor-β1(TGF-β1),the concentration of Gal-3 was detected by Western blot.After Gal-3intervention,the proliferation indexes PCNA,FGF2 and collagen secretion index Col-I of fibroblasts were detected by Western blot.The cell proliferation activity was detected by CCK-8.(2)An arraystar LncRNA gene chip sequencing of Gal-3 stimulated PAFs was detected and the differentially expressed LncRNA COMP was screened for q PCR verification.After overexpression of Gal-3 and knockdown of LncRNA COMP,the concentrations of its downstream protein COMP,fibroblast proliferation index PCNA and collagen secretion index Col-I were detected by Western blot.The expression of Col-I in PAFs was detected by immunofluorescence.The cell proliferation was detected by CCK-8.Cell migration was detected by scratch test.The concentration of COMP in lung tissue of MCT rats was detected by Western blot.(3)The concentrations of COMP,AKT and p-AKT in PAFs were detected by Western blot after stimulation of TGF-β1.si RNA of COMP and Akt activator SC79 were used to stimulate cells.The concentrations of fibroblast proliferation index PCNA and collagen secretion index Col-I were detected by Western blot.The expression of Col-I in PAFs was detected by immunofluorescence.The cell proliferation was detected by CCK-8.Cell migration was detected by scratch test.Results:(1)The concentration of Gal-3 in serum of PAH patients was higher than healthy people and there was a strong correlation between Gal-3 and NT-pro BNP;In the animal model,the concentrations of Gal-3 in the lung tissue of MCT rats were higher than normal rats.The concentrations of Gal-3 was increased after the stimulation of TGF-β1,and Gal-3 were positively correlated with cell proliferation indexes PCNA,FGF2 and collagen secretion index Col-I.(2)An arraystar LncRNA gene chip sequencing on Gal-3 stimulated PAFs was detected and the differentially expressed LncRNA COMP was screened for q PCR verification.Overexpression of Gal-3 at the cell level can increase the expression of LncRNA COMP and its downstream protein COMP,increase PNCA and Col-I,and increase the cell migration.Knockdown of LncRNA COMP can reverse PAFs migration and collagen secretion.The expression of comp in lung tissue of MCT rats was increased.(3)The concentration of Gal-3 and p-AKT/AKT was increased after stimulation of TGF-β1.The activation of Akt pathway led to the increased expression of Col-I,and cell migration.Knockdown COMP can inhibit the over activation of Akt pathway and inhibit the collagen secretion and cell migration of PAFs.Conclusion: Gal-3 up-regulates the expression of LncRNA COMP in PAFs,leading to the increase of the synthesis of COMP,activating the AKT pathway,enhancing PAFs cell migration and collagen secretion,and promoting the activation of PAFs in PAH. |
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