| ObjectivesThe model of acute lung injury induced by paraquat in rats was studied in this study,and the aim of this study was to investigate the effects of different dosage of hydrogen sulfide on the expression of LC3 and Beclin1 in autophagy in lung tissue.Methods30 healthy and clean SD male rats after 1 week of feeding,were randomly divided into normal saline control group(group N,0.9% 2ml/kg normal saline),paraquat poisoning model group(group PQ,paraquat solution 100mg/kg),low dose H2 S intervention model group(group L,100 mg /kg +Na HS 0.78mg/kg paraquat solution),middle dose H2 S intervention model group(group M,paraquat solution100mg/kg + Na HS 1.56mg/kg),high dose H2 S intervention model group(group H,100mg/kg + Na HS 3.12mg/kg of paraquat solution),CSE inhibitor PPG intervention model group(group P,paraquat solution 100mg/kg + PPG 30mg/kg),each group of 5rats.According to the experimental group,saline and paraquat solution were given group rats.After 3 hours,the groups were given saline,Na SH,PPG by intraperitoneal injection.After 24 hours,all the rats were sacrificed and the pathological changes of lung tissues were observed under electronic microscope,using BCA method to detect the content of bronchoalveolar lavage fluid.weighing it and calculating the W/D,using enzyme linked immunosorbent assay to determin the levels of Interleukin 10 and tumour necrotic factor-?,and Western Blot were used to detect the expression of autophagy protein LC3 II,LC3I and Beclin1.Result1.Pathological changes of lung tissueGeneral view: the lung tissue of group N rats seemed pink,smooth shape,no bleeding and edema,the lung tissue of group PQ rats seemed dark red,showing significant bleeding lesions,group L,group M,group H,the appearance of lung tissue compared with the group PQ,the degree of congestion and edema reduced in turn,the lung tissue of group P compared with that of group PQ showed more serious damage,showing large tracts of fresh bleeding lesions,severe pulmonary congestion and edema.Microscopic observation: the alveolar structure of group N rats is complete,there is no widening of interstitial lung,no leakage of red blood cells and inflammatory cell infiltration in alveolar and interstitial lung,no edema and hyaline membrane formation,and there is no atelectasis.In group PQ,the alveolar structure was damaged incompletely,showing obvious alveolar edema and pulmonary stroma was thickened,there were a large number of red blood cells and granulocytes infiltration,a large number of transparent film formation and large area of atelectasis.Compared with group N,the lung injury score of rats in this group was significantly higher(P<0.01).Compared with group PQ,there was no significant improvement in lung injury in group L and no significant decrease in lung injury score(P >0.05).Compared with group PQ,the alveolar and pulmonary interstitial edema in group M and group H decreased,the red blood cell and granulocyte infiltration decreased significantly,the area of the transparent membrane and atelectasis decreased,and the lung injury score decreased significantly(P <0.01).Compared with group PQ,the alveolar structure of group P was damaged more seriously,and large area alveolar edema,massive cell infiltration,thickening of pulmonary interstitium,large number of hyaline membrane formation and large area atelectasis,and pulmonary injury score were statistically significant(P <0.05).2.The changes of Lung wet / dry weight ratio(W / D)Compared with group N,the W/D value of group PQ was significantly increased,which was statistically significant(P <0.01).Compared with group PQ,the W/D value of group L was significantly lower(P <0.05).Group M and group H compared with group PQ,W/D values were significantly lower,with statistical significance(P<0.01).Compared with group PQ,the W/D value of group P was significantly increased,with statistical significance(P <0.05).3.The changes of IL-10 and TNF-aThe change of IL-10 value: Compared with group N,the IL-10 value of group PQ was significantly increased,with statistical significance(P <0.01).Compared with group PQ,the IL-10 content in group L was not significantly higher(P >0.05).The plasma IL-10 levels of group M and group H were significantly higher than those in group PQ(P <0.01).Compared with group PQ,the plasma IL-10 level of group P had no significant change(P >0.05).The change of TNF-avalue: Compared with group N,the TNF-a value of group PQ was significantly increased,with statistical significance(P <0.01).Compared with group PQ,TNF-a value of group L was not significantly reduced,there is no statistical significance(P >0.05).Compared with group PQ,TNF-a values of group M and group H were significantly lower,with statistical significance(P <0.01).Compared with the group PQ,the TNF-a value of group P was significantly increased,with statistical significance(P <0.01).4.The change of BALF protein concentrationCompared with group N,the BALF concentration of group PQ increased significantly(P <0.01).Compared with group PQ,the BALF concentration of group L was not significantly decreased(P >0.05).Compared with group PQ,the BALF concentration of group M and group H were significantly decreased(P <0.01).The BALF concentration of group P was significantly higher than that of group PQ(P<0.01).5.The changes of autophagy protein LC3 and Beclin1 expression in lung tissueCompared with group N,the expression of LC3-?,LC3-?and Beclin1 protein in group PQ were significantly increased(P <0.01).Compared with group PQ,the expression of LC3-?protein was significantly decreased in group L(P <0.05).The expression of LC3-?protein in group M and group H was significantly lower than that in group PQ(P <0.01).Compared with group PQ,the expression of LC3-?protein in group P was significantly increased(P <0.05).Compared with group PQ,the expression of LC3-? protein in group L was not significantly decreased(P >0.05).The expression of LC3-? protein in group M and group H was significantly lower than that in group PQ(P <0.01).Compared with group PQ,the expression of LC3-? protein in group P was significantly increased(P <0.01).Compared with group PQ,the expression of Beclin-1 protein in group L was not significantly decreased(P >0.05).The expression of Beclin-1 protein in group M and group H was significantly lower than that in group PQ(P <0.01).Compared with group PQ,the expression of Beclin-1 protein in group P increased significantly(P<0.05).Conclusion1.In the acute lung injury caused by paraquat,the autophagy of lung tissue was aggravated;2.Hydrogen sulfide can inhibit autophagy in acute lung injury in rats to and protect it.The mechanism may be related to down-regulation of autophagy protein LC3 and Beclin1. |
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