| BackgroundThe research of the research group showed that the integrin/integrin ligase/PI3 K signaling pathway may mediate the synthesis and degradation of extracellular matrix(ECM),and the integrin/integrin ligase/PI3 K signaling pathway may be a pathway for silicosis.One;studies have shown that the synthesis and degradation of ECM are involved in the occurrence and development of silicosis fibrosis,but the effect of Echistatin/BYL-719 intervention in the occurrence of silicosis fibrosis is less.Therefore,this study chose two inhibitors of this pathway(Echistatin/BYL-719)to explore the possible occurrence and development of silicosis.ObjectiveThe silicosis rat model was constructed by inhalation tracheal instillation,and Echistatin/BYL-719 intraperitoneal injection was used to intervene to explore the changes of related indicators before and after the intervention in rats,and analyze the changes of extracellular matrix components in the occurrence and development of silicosis fibrosis in rats In the role.Methods48 SPF male rats were selected and divided into blank control group,model group,Echistatin intervention agent group,BYL-719 intervention agent group,28 d and 56 d for each group,a total of eight groups,each with 6 rats.During the experiment,the body weight of the rats was measured at a fixed time every week.The rats in the 28 d and 56 d subgroups were anesthetized with chloral hydrate on 28 days and 56 days,weighed,and blood was collected from the abdominal aorta,and the rats were sacrificed.The lower lobe of the right lung was fixed with 4% paraformaldehyde for pathological observation;themiddle lobe of the right lung was cut on ice with a size of approximately 2 x 2 x 2 mm and fixed with 2.5% glutaraldehyde for electron microscope observation;left Lungs are stored in a refrigerator at-80 degrees for testing indicators;Western blotting is used to detect silicosis fibrosis indicators(type I collagen,type III collagen),extracellular matrix indicators(laminin(LN),bone bridge)in left lung tissue Protein(OPN)),extracellular matrix regulatory enzyme indicators(matrix metalloproteinase 1(MMP1),matrix metalloproteinase 3(MMP3))and other protein expressions.Result1.Observation of general symptoms of poisoning: One week after the model was established,the rats had no obvious abnormal symptoms.From the second week,the model group and the two intervention agent groups developed asthma,and pulmonary breath sounds could be heard.The model group and the two intervention agent groups had fatigue,laziness,no appetite,and hair loss.As time progressed,the symptoms above the rats in the model group gradually worsened,and the symptoms above the two intervention groups were slightly milder than those in the model group.At the time of sacrifice and autopsy,individual animals had lung cysts.2.HE staining pathological results: 28 days: the alveolar structure of the lung tissue of the rats in the saline group was normal without thickening,but there was slight inflammation in the alveolar interstitium;there were cellular nodules or silicic nodules in the lung tissue of the silicosis model group,And accompanied by a large number of inflammatory cell infiltration,the alveolar wall thickened to varying degrees,with the nodules most prominent.56 days: The general situation is the same as that of each group on 28 days;inflammatory cell infiltration is more obvious;the number of silicon nodules increases.In the Echistatin disintegrin group and BYL-719 PI3 K inhibitor group,the degree of fibrosis was reduced,the number of silicosis was reduced,but there were still inflammatory cell infiltration and alveolar space widening.3.During the feeding process,the silicosis model group and the two interventionagent groups had a certain decrease in dietary water consumption compared with the normal saline group,and their state was not as active as the normal saline group,and their weight decreased.Compared with the silicosis model group,the two intervention groups had a certain degree of improvement in weight loss.4.According to the analysis of variance,28 days after administration,the difference in the content of type I collagen in the lung tissues of the experimental groups was statistically significant(F=28.782,P<0.05).The silicosis model group and the normal saline group were significantly different.The content of type I collagen in the lung tissue of rats increased(P<0.05).Compared with the silicosis model group in the disintegrin intervention group and PI3 K inhibitor intervention group,the content of type I collagen in the lung tissue of rats decreased(P<0.05));The difference in the content of type III collagen in the lung tissue of the experimental groups was statistically significant(F=4.416,P<0.05),the silicosis model group was compared with the normal saline group,the content of type III collagen in the lung tissue of the rat Compared with the silicosis model group,the disintegrin intervention group and PI3 K inhibitor intervention group showed a decrease in the content of type Ⅲ collagen in the lung tissue of rats(P<0.05).5.According to the analysis of variance,28 days after the administration,the differences in the content of osteopontin in the lung tissues of the experimental groups were statistically significant(F=1.253,P<0.05).The silicosis model group was compared with the normal saline group.The content of osteopontin in the lung tissue increased(P<0.05).There was no significant difference in the content of osteopontin in the lung tissue between the disintegrin intervention group and the silicosis model.The PI3 K inhibitor intervention group was compared with the silicosis model group,The content of osteopontin in the lung tissue of rats increased(P<0.05);the difference in the content of laminin in the lung tissue of the experimental groups was statistically significant(F=2.072,P<0.05),the silicosis model group was compared with Compared with the normal saline group,the content of laminin in the lung tissue of rats increased(P<0.05).There was nosignificant difference in the content of laminin in the lung tissue of the rats compared with the disintegrin intervention group and the silicosis model(P<0.05)),compared with the PI3 K inhibitor intervention group and the silicosis model group,the content of laminin in the lung tissue of rats was reduced(P<0.05).6.According to the analysis of variance,28 days after the administration,there was a statistically significant difference in the content of MMP1 in the lung tissue of the experimental groups(F=5.754,P<0.05).The silicosis model group was compared with the normal saline group.The content of MMP1 in the lung tissue increased(P<0.05).Compared with the silicosis model group,the disintegrin intervention group and PI3 K inhibitor intervention group showed that the content of MMP1 in the lung tissue of rats increased(P<0.05);There was a statistically significant difference in the content of MMP3 in the tissues(F=6.371,P<0.05).Compared with the normal saline group,the MMP3 content in the lung tissue of rats increased in the silicosis model group(P<0.05).The disintegrin intervention group and Compared with the PI3 K inhibitor intervention group and the silicosis model group,the levels of MMP3 in the lung tissues of rats increased(P<0.05).7.According to the analysis of variance,56 days after the administration,the difference in the content of type I collagen in the lung tissue of the experimental groups was statistically significant(F=40.273,P<0.05).The silicosis model group and the saline group were significantly different.The content of type I collagen in the lung tissue of rats increased(P<0.05).Compared with the silicosis model group in the disintegrin intervention group and PI3 K inhibitor intervention group,the content of type I collagen in the lung tissue of rats decreased(P<0.05));The difference in the content of type Ⅲcollagen in the lung tissue of the experimental groups was statistically significant(F=7.438,P<0.05),the silicosis model group was compared with the saline group,the content of typeⅢ collagen in the lung tissue of the rat Compared with the silicosis model group,the disintegrin intervention group and PI3 K inhibitor intervention group showed a decrease inthe content of type Ⅲ collagen in the lung tissue of rats(P<0.05).8.According to the analysis of variance,after 56 days of administration,there was a statistically significant difference in the content of osteopontin in the lung tissue of the experimental groups(F=2.135,P<0.05).The silicosis model group was compared with the normal saline group.The content of osteopontin in the lung tissue was increased(P<0.05).Compared with the silicosis model group in the disintegrin intervention group and PI3 K inhibitor intervention group,the content of osteopontin in the lung tissue of rats increased(P<0.05);There was a statistically significant difference in the content of laminin in the lung tissue of rats in the experimental groups(F=4.077,P<0.05).Compared with the silicosis model group and the saline group,the content of laminin in the lung tissue of rats increased(P<0.05).Compared with the silicosis model in the disintegrin intervention group and PI3 K inhibitor intervention group,the content of laminin in the lung tissue of rats was reduced(P<0.05).9.According to the analysis of variance,after 56 days of administration,the differences in the content of MMP1 in the lung tissues of the experimental groups were statistically significant(F=11.491,P<0.05).The silicosis model group was compared with the normal saline group.The content of MMP1 in the lung tissue increased(P<0.05).Compared with the silicosis model group,the disintegrin intervention group and PI3 K inhibitor intervention group showed that the content of MMP1 in the lung tissue of rats increased(P<0.05);There was a statistically significant difference in MMP3 content in tissues(F=8.547,P<0.05).Compared with the silicosis model group and the normal saline group,the MMP3 content in the lung tissues of rats increased(P<0.05),the disintegrin intervention group Compared with the PI3 K inhibitor intervention group and the silicosis model group,the levels of MMP3 in the lung tissues of rats increased(P<0.05).Conclusion1.Echistatin/BYL-719 intervention can degrade the extracellular matrix of silicosis inrats;2.Echistatin/BYL-719 intervention can affect the regulation of extracellular matrix by MMP1 and MMP3;3.Echistatin/BYL-719 intervention can reduce the degree of pulmonary fibrosis. |
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