Effects Of Hepatopulmonary Syndrome Serum On The Myogenic Differentiation.proliferation And Migration Of Human Pulmonary Fibroblasts

Hepatopulmonary syndrome(HPS),a syndrome of serious chronic liver disease-derived pulmonary microvascular disease,the main feature is that " pulmonary microvascular dilation,gas exchange and hypoxemia ",it occurred mainly in patients with advanced cirrhosis.Pulmonary vascular remodeling(PVR)may be one of the main pathogenesis;the pathophysiological process is a interaction results of complex networks by many kinds of cells,appearance of smooth muscle like cells in non-muscle vascular site,muscular membrane and adventitial thickening,intimal plaque formation,resulting in pulmonary PVR;Of those,the accumulation of a large number of pulmonary artery smooth muscle cells(PASMCs)in the intima which is the core of change.Myofibroblasts(myofibroblast,MF)is a kind of special fibroblasts,the ultrastructural features intermediate between fibroblasts and PASMCs,is a kind of PASMCs like cells.CBDL serum promote PASMCs phenotypic conversion from contraction type(high differentiation)to the undifferentiated synthesis type and migration,and is the basic pathological changes of PVR.Recently more and more studies find that adventitial fibroblasts can differentiate into MF in the presence of hypoxia.We hypothesized that the activation of MF may migrate into the intima and abnormal proliferate,directly involved in the accumulation of a large number of PASMCs in endometrial in the process of PVR.However,the mechanism is not clear.Early we have studied the role of hepatopulmonary syndrome rat serum in proliferation,migration and phenotype conversion of rat PASMCs,the result is HPS rat serum can promote PASMCs conversion to synthetic phenotype,and enhance the ability of cell proliferation and migration.many cytokine produced bypathologic liver through the blood circulation to adventitia LFs of lung,which caused pathological pulmonary vascular remodeling,this is the main purpose of this study.In this study,we observed myogenic differentiation,proliferation and migration of human LFs.Methods:1.culture and identification of human primary lung LF,LF were passage cultured after 3-4 days and were detected using light microscopy,immunofluorescence assay.2.HPS serum were collected according to the inclusion criteria,selected qualified HPS patients and healthy controls,and a small sample was extracted from peripheral blood serum.3.respectively treatment of LFs with normal and hepatopulmonary syndrome patients serum,immunohistochemical detected the expression of SM-alpha –actin in LFs.4.respectively treatment of LFs with normal and hepatopulmonary syndrome patients serum,Western blot detected the expression of SM-alpha –actin and SM-MHC in LFs..5.respectively treatment of LFs with normal and hepatopulmonary syndrome patients serum,3H-Td R and transwell assay detected the ability of proliferation and migration in LFs..Results:1.Under the inverted microscope,primary cultured human fibroblast was monolayer growth and palisade arranged radially,cell morphology were long spindle shaped,with thick processes,abundant cytoplasm,the nucleus is clear oval.The immunofluorescence staining of vimentin was positive in Fibroblast.2.HPS patient's serum were collected,stored for-80.3.immunohistochemistry results,compared with normal serum group,HPS patients serum cultured cells were rod-shaped and nucleus were oval shaped,a large number of brown particles in cytoplasm.Under inverted microscope,normal serum stimulated cells were monolayer growth,like flies or triangle shape,clear nucleus,abundant cytoplasm,,immunohistochemical staining of SM a-actin was no obvious brown yellow precipitate in the cytoplasm.4.western bolt showed that,compared with normal serum,SM-alpha-actin and SM-MHC protein expression were increased in HPS serum group.And the expression of SM--actin and SM-MHC protein were increased significantly with the extension of the time,especially in 48 h and 72 h,(P<0.05).5.3H-TdR and Transwell assay detected,HPS patients serum can enhance the ability of proliferation and migration in LF in vitro significantly,especially in 48 h and 72 h.Conclusion:1.Hepatopulmonary syndrome patients serum can promote myogenic differentiation of human LF in vitro.2.Hepatopulmonary syndrome patients serum can enhance the proliferation and migration ability of human LF.