The Expression Of ADAMTS-1in Pulmonary Fibrosis Tissue Of Rats And The Intervention Of Oxymatrine

Objective: To detect the dynamic changes of pulmonary ADAMTS-1expression indifferent periods of rats with bleomycin-induced pulmonary fibrosis, and effects ofoxymatrine on ADAMTS-1expression, thereby clarifying the molecular mechanisms bywhich ADAMTS-1is involved in pulmonary fibrosis and oxymatrine suppresses pulmonaryfibrosis.Methods: Sixty rats were randomly divided into four groups:control group（n=15）groupand the intervention group were sacrificed on7,14, and28days, model group (n=15)、thelow-dose (50mg/kg) Oxymatrine intervention group(n=15)，the high-dose (100mg/kg)oxymatrine intervention group (n=15).5rats from each group were sacrificed on7、14、28days.Pathological changes of pulmonary fibrosis were determined by hematoxylin eosinstain and Masson stain. Expression levels of pulmonary ADAMTS-1protein and mRNAwere detected through immunohistochemical method and RT-PCR, respectively.Results:1.hematoxylin eosin stain and Masson stainHE stain:the rat alveolar structure of control group was normal, clear, there are noalveolar septa widened and abnormal collagen deposition. On7days, the model rats showedobvious acute inflammation, and there are many inflammatory cells in the alveolar spaceand pulmonary interstitial. On14days, lung interval significantly widened, collagendeposition increased and fibrosis change with patchy areas formed. On28days,A lot ofcollagen fibers hyperplasied, the pulmonary tissue structure disorder.there was a littleinflammatory cell infiltrated and the stable pulmonary fibrosis has formed. There weresignificantly difference between every group.Masson staining:Stromal collagen showing ablue-green,at14days and28days a large number of collagen fibers gathered and deposited.There was scattered collagen fibers in control rats’ bronchial wall and pulmonary interstitial. The distribution and quantity of collagen fibers in low-dose Oxymatrine intervention groupwas less than that in model group. Compared with the low-dose intervention group andmodel group, the aggregation of the collagen fiber in high-dose oxymatrine interventiongroup was significantly eased.. Masson stain: Stromal collagen showing a blue-green,at14days and28days a large number of collagen fibers gathered and deposited. There wasscattered collagen fibers in control rats’ bronchial wall and pulmonary interstitial. Thedistribution and quantity of collagen fibers in low-dose Oxymatrine intervention group wasless than that of collagen fibers in model group. Compared with the low-doseintervention group and model group,the aggregation of the collagen fiber in high-doseoxymatrine intervention group was significantly eased.2.HYP content determinationWith the time of constructing pulmonary fibrosis model, the content of HYP becamehigher. From14days the HYP content of the model group increased gradually.The HYP ofthe model group is much higher than control group,low-dose oxymatrine group andhigh-dose oxymatrine group(P <0.05, P＞0.05).3.the expression of ADAMTS-1in pulmonary fibrosisADAMTS-1was expressed in both model group and oxymatrine intervention groupon7,14,28days.Its expression level decreased gradually with the time increment. Theexpression of ADAMTS-1in oxymatrine intervention group was higher than model group.Compared with control group,the expression of ADAMTS-1in model group wassignificantly decreased, and there were significant difference between them（P <0.01）.Compared with model group,the expression of ADAMTS-1in low-dose interventiongroup was increased, but there was no significant difference（P>0.05）. Compared withmodel group,the expression of ADAMTS-1in high-dose oxymatrine intervention groupwas significantly higher（P <0.05）. Compared with low-dose intervention group, theexpression of ADAMTS-1in high-dose oxymatrine intervention group was significantlyhigher, and there was significant difference（P <0.05）. Compared with control group,theexpression of ADAMTS-1in low-dose intervention group was significant difference（P<0.05）. 4.4.Detecting the expression of ADAMTS-1and COLI mRNA inPulmonary fibrosis by RT-PCROn7days,14days,28days,ADAMTS-1mRNA expression in all groups, andespecially in control group. Expression were significantly different among the groups andtime points (P <0.05),except the result of model group and low-dose intervention group’scomparation. COLI mRNA were expressed at7days,14days,28days of rats’pulmonaryfibrosis, Compared with control group, which expressed in each group and each point-intime all have significantly differen（tP <0.05）,and that the expression of COLI mRNA in themodle group is the highest at28days.Conclusion:1.The expression of ADAMTS-1is up-regulated in the pulmonary tissues of rats withpulmonary fibrosis.2. The levels of pulmonary COLI are decreased in rats with pulmonary fibrosis.3.Oxymatrine can inhibit pulmonary fibrosis through diminishing COLI content viapromoting the expression of ADAMTS-1.