The Effect Of The Tumour Necrosis Factor-alpha (TNF-a) Combining The Prostaglandins E₂(PGE₂) And TNF-a Combining The Oxymatrine On The Signaling Material Of Pulmonary Fibroblast

Objective:To study the effect of the tumour necrosis factor-alpha(TNF-α)combining the prostaglandins E₂(PGE₂) and TNF-α combining the oxymatrineon the signaling material of pulmonary fibroblast.To probe the mechanism ofpulmonary fibrosis from the facet of signal conduct and provide the evidence forlook for the target of molecular to treat pulmonary fibrosis.Methods:SDneonatal rats' pulmonary fibroblasts were cultured,and were subcultured by 4-5generations. The pulmonary fibroblasts were stimulated by the differentconcentration TNF-α,TNF-α combined different concentration PGE₂ and TNF-αcombined different concentration oxymatrine.1.Measure the content of (inositol1,4,5-trisphosphate(IP₃),cyclic adenosine monophosphate(cAMP), cyclicguanosine monophosphate(cGMP) of pulmonary fibroblast by means ofradioimmunoassay(RIA).2.Measure the calcium concentration of pulmonaryfibroblast by flow cytometry(FCM).3.The expression of TNF-α receptor ofpulmonary fibroblast by means of immunohistochemistry.Results: 1.Whendifferent concentration of TNF-α actting on the pulmonary fibroblast,thepositive rates of TNF-αR,the IP3cpm and the concentration of calcium alldisplayed increasing trend with the dose of TNF-α increasing ;compared withother groups,control group was obviously significantly(P<0.05);the picture offlow cytometry (FCM) of calcium of 10μg/L TNF-α group (blue picture)moved to right significantly;when TNF-α actting 4 hours,compared with othergroups the value of cAMP and the ratio of cAMP/cGMP of 10μg/L TNF-αgroup was higher significantly(P<0.05),but when actting 24 hours these lowerthan other groups.2.When TNF-α and PGE₂ actting the pulmonary fibroblasthomochronously, compared with other groups the positive rates of TNF-α,theIP3cpm(when it on 30s and 60s) and concentration of calcium of 10μg/LTNF-α group was higher significiantly; compared with other groups the pictureof FCM of 2.0μg/L PGE₂ group moved left signiciantly;the value of cAMPdisplayed increasing earlier while descend latter;the value of cAMP of 10μg/LTNF-α group descend signicantly(P<0.05);when actting 4 hours, the ratio ofcAMP/cGMP of 10μg/L TNF-α associated with 0.5μg/L PGE₂ group washigher than 10μg/L TNF-α group significiantly the ratio of cAMP/cGMP of allgroups of 4 hours were higher than those of 24 hours.3.When TNF-α andoxymatrine (OM) actting on the pulmonary fibroblasthomochronously;compared with other groups, the positive rates of TNF-αR andconcentration of calcium of 10μg/L TNF-α group was higher signiciantly;thepicture of FCM of 800mg/L OM group(blue picture) moved to leftsignificiantly,the value of cAMP and the ratio of cAMP/cGMP displayedincreasing trend with the dose of OM increasing and they got to the top in800mg/LOM group.Conclusion: 1. When the TNF-α combined its receptor acton pulmonary fibroblast, this actting may be by means of changing the secondsignaling messenger of the signaling pathway mediated by GPCR.2.ExtrinsicPGE2 may be a new medicine treat pulmonary fibrosis and its mechnism may bechange the second signaling material by means of inhibit the TNF-α to combineits receptor .3. The mechnism of oxymatrine inhibit pulmonary fibrosis may beaccomplished by means of inhibit the combination of TNF-α and its receptorand cause the changing of the second signaling material.4. The target of treatpulmonary fibrosis may be control TNF-α combine its receptor.