| Objective: To observe the effect of reduced glutathione(GSH)on the gene expression of pulmonary surfactant in acute lung injury(ALI)mice induced by agkistrodon halys venom,and to explore the protective effect of GSH on acute lung injury induced by agkistrodon halys venom.methods:1.Group: 60 Kunming mice(40 5g)were randomly divided into six groups: group A(0.9% ns intraperitoneally),group B(1800mg / kg GSH intraperitoneally),group C(0.75 mg / kg Agkistrodon venom intraperitoneally),group D(0.75 mg / kg Agkistrodon venom intraperitoneally + 1800 mg / kg GSH),group E(1.0mg / kg Agkistrodon venom intraperitoneally),group F(1.0mg / kg Agkistrodon venom intraperitoneally + GSH)1800mg/Kg)?2.To establish the Ali model of mice induced by agkistrodon halys venom:(1)in addition to group A and B,the other four groups were intraperitoneally injected with different doses of Agkistrodon halys freeze-dried powder solution,and group A and B were intraperitoneally injected with equal volume of 0.9% ns.(2)Three hours later,1800 mg / kg of reduced glutathione was injectedintraperitoneally in groups B,D and F respectively,and 0.9% NS was injected intraperitoneally in groups A,C and E.(3)After injection of glutathione for 5hours,the experimental animals in each group were killed and lung samples were collected.3.To determine the detection index of experimental design:(1)to make paraffin section of lung tissue and he staining,observe the pathological changes under optical microscope and pathological score of lung injury;(2)to prepare lung tissue homogenate and measure the activity of GSH Px in lung homogenate;(3)to measure the expression level of SP-A,SP-B and SP-C mRNA in lung homogenate by fluorescence quantitative PCR.Results:1.Pathological changes of lung tissue and pathological score of lung injury(IQA): Compared with the lung tissue of group A,there was no significant difference in IQA of group B(P > 0.05);IQA of group C,D,E and F was significantly higher(P < 0.01).In group C,The structure of pulmonary alveoli is disordered and destroyed,the pulmonary interstitium is thickened,the alveoli are edematous,part of the alveolar hyaline membrane is formed,the red cells in the pulmonary interstitium and alveoli leak out and dissolve,and a large number of inflammatory cells infiltrate.Compared with group C,the damage of lung tissue in group D was alleviated,and the alveolar structure was basically intact,but the alveolar septum was thickened,inflammatory cells infiltrated and IQA decreased significantly(P < 0.01).In Group E,lung injury was the most serious,alveolar collapse and severe atelectasis,extensive hyaline membrane was formed,a large number of red blood cells leaked out and dissolved,inflammatory cells infiltrated,and a large number of neutrophils gathered into clusters,macrophages showed patchy aggregation,IQA significantly increased(P < 0.01).Compared with group E,there was no significant improvement in lung injury in group F and no significant difference in IQA(P > 0.05).The score of lung injury in mice with ALI induced by agkistrodon halys Pallas venom after intraperitoneal injection of reduced glutathione.2.Changes of GSH Px activity in lung homogenate: compared with group A,GSH Px activity in lung homogenate in group B increased(P < 0.01),while GSH Px activity in lung homogenate in group D,E and F increased significantly(P < 0.01);GSH Px activity in lung homogenate in group C had no significant difference(P > 0.05).Compared with group C,GSH Px activity in group D increased(P < 0.05).There was no significant difference between group E and group F(P > 0.05).GSH Px activity increased after GSH injection in mice.3.The expression level of SP-A,SP-B and SP-C mRNA in lung tissue homogenate: compared with the blank control group A,there was no significant difference in the expression level of SP-A,SP-B and SP-C mRNA in lung tissue of group B(P > 0.05);the expression levels of SP-A,SP-B and SP-C mRNA in lung tissue of group C and E were all decreased(P < 0.05).Compared with low dose Agkistrodon C group,the expression of SP-A,SP-B and SP-C mRNA in lung tissue of high dose Agkistrodon E group decreased(P < 0.05).Comparedwith group C,the expression levels of SP-A,SP-B and SP-C mRNA in lung tissue of group D were significantly higher(P < 0.01).Compared with group E,the expression level of SP-A and SP-B mRNA in lung tissue of group F was significantly higher(P < 0.01),but there was no significant difference in the expression level of SP-C mRNA(P > 0.05).The expression levels of SP-A,SP-B and SP-C mRNA in lung tissue of mice with acute lung injury induced by agkistrodon halys venom were all decreased,and the more serious the poisoning was,the more obvious the decrease was.The expression of SP-A,SP-B and SP-C mRNA in lung tissue of mice with acute lung injury induced by agkistrodon halys Pallas venom was increased after injection of reduced glutathione.However,the expression of SP-A and SP-B mRNA in lung tissue of mice with acute lung injury induced by agkistrodon halys venom was only increased after injection of reduced glutathione.Conclusions:1.The expression levels of SP-A,SP-B and SP-C mRNA in lung tissue of mice with acute lung injury induced by agkistrodon halys venom were all decreased,and the more serious the poisoning was,the more obvious the decrease was.2.Glutathione can enhance the activity of GSH Px,reduce the pathological score of lung tissue,reduce the lung injury,and enhance the expression of SP-A,SP-B and SP-C mRNA in lung tissue.3.The activity of GSH PX in lung tissue of mice with acute lung injuryinduced by agkistrodon halys Pallas venom was increased.The expression of SP-A and SP-B mRNA was increased after injection of reduced glutathione,but the lung injury could not be reduced... |
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