Inhibitory Effect Of Peroxiredoxin-1 On Inflammation Formation In Mice With Silicosis

Objectives Peroxiredoxin?Prx-1?is a novel peroxidase.In this study,lentiviral vectors transfection was used to inuce Prx-1 overexpress in lung tissue of mice.The aim is to explore the effect of Prx-1 on SiO₂-induced inflammation formation indeuced by silicosis,and the possible mechanism was also been found.The study would provide a new research direction for the new targets of silicosis target therapy.Methods Twelve weeks,male,C57 mice were randomly divided into 4 groups?n=10?:1)control group:normal saline;2)silicosis group:bronchial intubation to give SiO₂ perfusion?5mg/mouse?;3)transfection control group:bronchial intubation to give SiO₂?5mg/mouse?+empty lentivirus?10⁷TU/mouse?;4)Prx-1 transfection group:bronchial intubation to give SiO₂?5mg/mouse?+Prx-1 lentivirus?10⁷TU/mouse?.All mice were raised normally for one week,and then they were sacrificed.H.E staining was used to observe the morphological changes of lung tissues;immunohistochemistry was used to detect the expression of 8-OHdG,CD68,M1 macrophages?iNOS?;Finepointe WBP whole body plethysmography system was used to detect lung function;Western blotting was used to detect the expression levels of Prx-1,NF-?B and P-Src proteins;HE staining was used to count the number of macrophages in the alveolar lavage fluid under high power field;and real-time PCR was used to detect IL-1?,TNF-?and TGF-?1.Results 1 H.E staining showed that the lung tissue of the control group was normal,with slender alveolar septa and little inflammatory cell infiltration;the lung tissue of the silicotic group and the transfected control group showed obvious silicotic nodules,with macrophage and lymphocyte infiltration in the alveolar space and alveolar septa;compared with the silicotic group and the transfected control group,the pathological changes of the lung tissue of the Prx-1 transfected group were reduced,and the silicotic nodules were smaller.2 The results of immunohistochemistry showed that the positive expression scores of 8-OHdG,CD68,and iNOS in the silicosis group and the transfection control group were significantly increased compared with the control group;the positive expression scores of8-OHdG,CD68,and iNOS,in the Prx-1 transfection group were significantly decreased compared with the silicosis group and the transfection control group.3 The results of pulmonary function showed that compared with the control group,the tidal volume?TVb?,minute ventilation volume?MVb?,maximum inspiratory flow rate?PEF?,and respiratory rate?f?were decreased in the silicosis group and the transfection control group;compared with the silicosis and transfection control group,the tidal volume?TVb?,minute ventilation volume?MVb?,maximum inspiratory flow rate?PEF?and respiratory rate?f?were increased and decreased in the Prx-1 transfection group.The macrophage count in alveolar lavage fluid showed that the number of macrophages increased in the control group compared with the silicosis group and the transfection control group,and decreased in the Prx-1 group compared with the silicosis group and the transfection control group.5Immunoblotting results showed that:compared with the control group,the expression of Prx-1,NF-?B and P-Src protein was up-regulated in the silicosis group and the transfected control group;compared with the silicosis group and the transfected control group,the Prx-1 protein expression was further up-regulated in the Prx-1 transfected group,and the expression of NF-?B and p-Src protein was decreased.6 Realtime PCR results showed that compared with the control group,the expression levels of IL-1?,TNF-?,and TGF-?1 in the silicosis group and the transfected control group were significantly upregulated;compared with the silicosis and transfected control group,the expression levels of IL-1?,TNF-?,and TGF-?1 in the Prx-1 group were significantly downregulated.Conclusions Overexpression of Prx-1 has an inhibitory role in silicosis-induced inflammatory response,mediated by repression of ROS/NF-?B/Src pathway activation,inhibition of macrophage polarization and synthesis of IL-1,NF-?.Figure 9;Table 2;Reference 47...