| Bleomycin(BLM)is a natural product of aminoglycosides.It was isolated from the culture medium of Streptomyces clostridium in 1966.It has been used for more than 50 years and has strong anticancer activity,especially for lymphoma,phosphorus cell carcinoma,lung cancer and reproductive system cancer has a good effect,while no significant bone marrow suppression,does not inhibit the body’s immune function,making it an important drug for clinical cancer chemotherapy,but its severe pulmonary fibrosis Toxic side effects,great restrictions on its clinical enough amount of course of treatment.Therefore,how to ensure the full amount of BLM adequate course of treatment based on the prevention and treatment of pulmonary fibrosis is the key to further improve the prognosis.Intergrin-linked kinase(ILK)is a new protein with protein kinase activity,which is identified by Hannigan et al.Using yeast two-hybrid technique.It has shown that it plays an important role in a variety of growth factor signaling pathways.Downstream of the target protein,and then regulate cell proliferation,growth,differentiation,migration,invasion and tumor angiogenesis and other processes.Epithelial-mesenchymal transition(EMT)is an important process of pulmonary fibrosis,and a number of studies have shown that ILK is closely related to EMT of cells and promotes the development of EMT.The activated ILK mediates the upregulation ofα-smooth muscle protein and is involved in the formation of EMT.Another recent study found that Wnt signaling pathway involved in multiple organ fibrosis process,and confirmed Wnt / β-catenin(β-catenin)signal transduction system is necessary for lung development of the molecular signaling system,The association is also closely related to the pathway involved in fibrosis,which is mainly associated with the nuclear aggregationof β-catenin.When β-catenin phosphorylation is inhibited,it can not be ubiquitin recognition,which can not be protease complex degradation,and then in the cytoplasm of a large number of aggregation and nuclear aggregation,leading to tissue fibrosis.Recent studies have found thatβ-catenin and ILK important dialogue: β-catenin nuclear aggregation and glycogen synthase 3β(Gkycogen synthase kinase-3beta GSK-3β)phosphorylation,while Virchows and other studies have shown that activated ILK also Can affect the phosphorylation of GSK-3β,so the two may have synergistic effect in pulmonary fibrosis.Pulmonary fibrosis is a progressive increase in interstitial lung disease,pathological manifestations of diffuse alveolitis,alveolar unit structural disorders and pulmonary fibrosis,the exact pathogenesis is still unclear.Generally due to chemical or other causes of pulmonary inflammation,followed by exudation,fibroblast proliferation and extracellular matrix protein deposition,eventually leading to irreversible lung injury,although there is no characteristic drug treatment,but a major breakthrough at home and abroad found that,Macrolide antibiotics can prevent the development of fibrotic diseases,and even improve lung function.He Kun and other studies have found that erythromycin can prevent bleomycin-induced pulmonary fibrosis.Niu Xiaojuan and other studies have found that azithromycin can reduce the pulmonary fibrosis rat model of collagen deposition,with dexamethasone similar role,and less adverse reactions.But macrolide antibiotic anti-fibrosis mechanism is still in the exploration.Objective:The aim of this study was to investigate the possible mechanism of azithromycin on pulmonary fibrosis by observing the content of ILK andβ-catein in the intervention group and model group,and to provide possible solutions for the clinical application of bleomycin-induced side effects.Methods:Select 100 healthy Kunming mice,weight 18-22 g,raised after one week,were randomly divided into group A model group(n = 20)and intracheal injection of BLM(3.5mg / kg,with 1 ml saline solution),in the first day after modeling saline 1ml / 10 g,1 / d,a total of 28 days;B group of control group:20,tracheal injection of 1ml saline,After the first day of saline 1ml gavage,1 /d,a total of 28 days.Group C was treated with azomycin and intraperitoneally injected with BLM(3.5 mg / kg,dissolved in 1 ml saline).on the first day after modeling azithromycin(1ml 5mg/Kg NS)orally,1 times / day,a total of14 days.Group D was treated with azomycin and intraperitoneally injected with BLM(3.5 mg / kg,dissolved in 1 ml saline).on the fifteenth day after modeling azithromycin(1ml 5mg/Kg NS)orally,1 times / day,a total of 14 days.Group E was treated with azomycin and intraperitoneally injected with BLM(3.5 mg / kg,dissolved in 1 ml saline).on the first day after modeling azithromycin(1ml 5mg/Kg NS)orally,1 times / day,a total of 28 days.The above groups were scarificed at 7,14,21,28,respectively,each of the 5.The right frontal femoral artery was sacrificed and the right lung homogenate was immediately sacrificed and placed in a refrigerator at-70 ℃.After the specimen was collected,the contents of ILK and β-catenin were determined by ELISA.Left lung tissue was placed in 4% formaldehyde fixative solution after HE staining and Masson staining to evaluate the degree of alveolar inflammation and pulmonary fibrosis.The experimental data were processed by SPSS17.0 statistical software package.The count data were expressed by(X ± SD).At the same time point,the same index comparison between groups to meet the normality and homogeneity of variance under the independent sample T test,does not meet the nonparametric test for independent samples of normal or homogeneity of variance under the condition of two variables related to the use of Spearman rank correlation analysis,P <0.05 as the test standard.Result:1 Lung histopathological analysis:At the same time point,A group of mice alveolitis degree and degree of fibrosis were significantly higher than B group.C group and E mice were mild pulmonary alveolar inflammation and fibrosis,two groups Compared with group A,the degree of alveolitis and fibrosis in mice were significantly reduced.In group D,the same degree of alveolar inflammation and fibrosis were observed at the same time point in group A and lung cancer on day 7 and day 14,and azithromycin was given The inflammation and fibrosis of lung tissue of group D were significantly reduced at the same time point as in group A on day 21 and day 28.The pathological results of each group showed that pulmonary fibrosis was divided into two stages: early inflammation and advanced fibrosis,which accorded with the pathological process of human pulmonary fibrosis,and the inflammatory reaction gradually decreased with the progress of fibrosis,and the degree of fibrosis gradually increased.2 The content of ILK and-catenin in lung tissue2.1Comparison between in different groups1)At the same the same time;A group of ILK,beta-catenin content in lung tissue of mice were significantly higher than group B,especially in the group A 28 day increases the most significant,the difference had statistical significance(P < 0.05);2)At the same time: the levels of ILK and β-catenin in lung tissue of group C were significantly lower than those in group A(P <0.05);3)AT the same time points: the contents of ILK,-catenin beta D group of lung tissue in mice at seventh days and fourteenth days than in the A group had no obvious change,there were no significant differences(P > 0.05);in twenty-first days,twenty-eighth days in the lung tissues of ILK,beta-catenin content in A group were significantly decreased,with the the difference statistically significant(P<0.05);4)The same time point: group E of lung tissue in mice ILK and beta-catenin.Compared with A group were significantly decreased,the differences were statistically significant(P <0.05);5)The same time point: group C of lung tissue in mice ILK and beta-catenin.Compared with E group did not change significantly,there were no significant differences;2.2Comparison between in same groups 1)The different time: A group of lung tissue in mice,ILK content of beta-catenin gradually increased,the differences were statistically significant(P<0.05);2)The different time points: B,C and E in lung tissue of three groups of mice ILK and beta-catenin content had no obvious change,there were no significant differences(P> 0.05);3)The different time points: the contents of ILK and beta-catenin in lungs of mice in D group were different in azithromycin before and after intervention,D group of mice lung tissue ILK and beta-catenin content in seventh days,fourteenth days and twenty-first days and 28 days were significantly lower,the differences were statistically significant(P<0.05).3 Correlation analysis of ILK and beta-catenin in 3 lung tissues:The contents of different time points in group A bleomycin model group lung tissue in mice ILK and beta-catenin were higher than group B blank group of mice lung tissue ILK and beta-catenin were significantly increased,two were positively related,with statistical significance(P = 0.001,r = 0.978).Conclusion:1 Azithromycin can block bleomycin induced pulmonary fibrosis in mice occurrence,progress,and could become a clinical solution with bleomycin tumor patients to prevention and treatment of pulmonary fibrosis.2 Azithromycin intervention effect of pulmonary fibrosis in mice is associated with drug use time and drug treatment,inflammation in the early pulmonary fibrosis stage,late pulmonary fibrosis in mice fibrosis stage also has certain effect.3 ILK,beta-catenin all participate in the process of pulmonary fibrosis,both exist positive correlation,suggest the possible synergies in the process of fibrosis. |
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