Detection The Express Level Of AKT?p-AKT Protein In Lung Tissue By The Experiment Of Rat That Hirudin Intervents To Idiopathic Pulmonary Fibrosis Caused By Bleomycin

Objective:Hirudin was chosed as study drug in this paper,mainly observed changing of IPF rats' general condition and Lung morphology,using HE and Masson to observe their lung morphology pathology changes,took Western-blot method detect the express level of AKT?p-AKT protein in rat lung tissue,discovered the function of hirudin cure idiopathic pulmonary fibrosis(IPF)and seek its mechanism for IPF.Methods:Selected 180 rats of equal age and weight from the SPF laboratory,and randomly selecting 18 rats from 180 rats as blank group(K group),rest 162 rats mold-ing IPF by bleomycin.Randomly choosed 108 rats from successful molding,based on random number table method,divided into model group(M),hormone(J),high-hirudin(G),middle-hirudin(Z)and low-hirudin(D)and middle-hirudin+hormone group(ZJ)with 18 rats in each group.Started to take medicine after 2 days success molding,respe ctively killed rats on 7th day,14 th day and 28 th day and collecting changing data of r ats' general condition and lung tissue,observe the data changing under HE and Mass on stain,took Western-blot method to detect the expression of AKT and p-AKT protein in rats' lung tissue.Result:1.Observe rats' general conditions Found the K group is good,M group rats' skin are dull and dry,their breathing are all rapid and shallow,and lost weight noticeably,compared to the K group,P<0.01.J group rats' mental state are fair at early stag ?e,finally fur is yellow and weight increase slower.Hirudin G group and ZJ group rats' breathed slightly smooth and gained weight rapidly compared to J group.Zgroup and D group rats' fur are yellow,less activity,rapid-shallow-breathing,put on weight slower than J group'.2.Observed appearance of rats' lung tissue K group rats' lung tissue was in nor mal,M group rats' appearance changing was coinciding with typical IPF pathological changes.The volume are significantly narrowed,uneven colored,the quality of a material become to harden.J group rats did not have a decide change in the early stage,gradually become slightly smaller,some tissues is consolidation.G group rats' volume reduced a little,the degree of consolidation are lighter than J group,Z group and D group rats' appearance equal to J group'.3.Observed results of histopathology after HE and Masson stain K group rats' histopathology are normal under high power microscope,As time progresses M group rats' disease have an number of inflammatory cell infiltration,alveolar interval widening,collagenous fiber and fibroblast proliferation formation,pulmonary interstitial fibrosis.Except for 14 d,the pulmonary fibrosis score was slightly lower than M group',P>0.05,and there was no statistical significance.At the other time points,the lung pathological damage and fibrosis score of J group' was lower than that of M group',P<0.01 or P<0.05,there was statistical difference.The score of ZJ group and G group was lower than that of J group' at different time.At different time points,the pathological damage and fibrosis score of lung tissue in Z group' and D group' were slightly higher than J group'.At different time points,the score of ZJ group was slightl y lower than G group'.4.The results of AKT protein expression In different time,K group rats' lung AKT protein levels are always minimum,at the 7th day and the 28 th day in M group rats' AKT protein content was obviously higher than J group,and P<0.01.at the 7th day G group rats' AKT protein content lower than J group',at the 14 th day and the28 th day,G group and ZJ group' lung AKT protein content lower than J group,and P<0.01,there was a significant statistical significance.At the 7th day and the 28 th day,Z group rats' lung AKT content was obviously higher than J group's,and P<0.01,there was a significant statistical significance.5.The expression results of p-AKT protein It reveals that,K group rats' lung p-AKT protein content is the lowest at three time points,M group' is highest,J group pAKTprotein content was lower than M group,and P<0.01,there are statistically significantdifference.ZJ group rats' lung p-AKT protein content was lower than J group's,and P<0.01,there were statistically significant difference.At the 7th day and the 14 th day Ggroup' p-AKT protein content below J group,and P<0.01.At the 28 th day G group'p-AKT protein content is higher than J group's,and P>0.05,but no statistical difference.Conclusion: Hirudin can intervene IPF by reducing the expression of AKT and p-AKT protein expression in lung tissue,PI3K/AKT signaling pathway is one of the mechanism of hirudin acting on IPF;intervention effect of hirudin on IPF with dose effect relationship;hirudin and hormones have synergistic effect in the process of intervention in IPF.