| Objective:To observe the effect of trioxygenated preconditioning on the morphology and function of rat pulmonary ischemia-reperfusion injured cells.Methods:30 SD rats(400-500 g,male and female)were randomly divided into three groups:trioxygenated preconditioning group(O3+I/R group,n=10),ischemia reperfusion group(1/R group,n=10)and sham operation group(S group,n=10).In the three-oxygen pretreatment group,three-oxygen body(50 ug/ml,1 mg/kg/d)was injected intraperitoneally at fi0xed time points every day for 7 consecutive days.The same volume(ml)of oxygen was injected at the corresponding time in the control and sham-operated groups.After anesthesia,all rats were anesthetized,trachea was intubated in the neck,ventilator was connected,and the left hilum was separated after thoracotomy.The sham-operated group was only threaded and not ligated after the free hilum.O3+I/R,I/R group:After freeing the hilum,4-0 silk suture ligation was used to ligate the hilum for 40 min and reperfusion for 100 min.After the perfusion was completed,the rats were sacrificed by cardiac bleeding.Use a syringe to extract the PBS buffer pre-cooled to 4? for alveolar lavage,slowly and repeatedly lavage 2-3 times,with an interval of no more than 2 minutes,and then withdraw the alveolar lavage fluid to complete the modeling.After the experiment,the rat lung tissues were taken out,2*2*2mm lung tissues were placed in osmium acid storage solution and fixed for electron microscopy observation,and some lung tissues were taken for determination of lung wet-dry specific gravity.LDH activity,expression of the pinkl gene,and the remaining part is placed in a refrigerator and stored frozen.Results:Compared with sham operation group,the W/D values of the three-oxygen preconditioning group and ischemia-reperfusion group were higher(P<0.05),and the W/D values of the three-oxygen preconditioning group and ischemia-reperfusion group were significantly higher than those of the three-oxygen preconditioning group(P<0.05).Compared with sham-operated group,ROS content in ischemia-reperfusion group and trioxygenated preconditioning group increased significantly(P<0.05),while ROS content in ischemia-reperfusion group and trioxygenated preconditioning group increased significantly(P<0.05).Compared with sham-operated group,LDH activity was significantly increased in ischemia-reperfusion group and trioxygenated preconditioning group(P<0.05),while LDH activity was significantly increased in ischemia-reperfusion group and trioxygenated preconditioning group(P<0.05).In the ischemia-reperfusion group,part of the mitochondrial crest of the tissue cells disappeared,the volume increased,some cells showed vacuolization,microvilli decreased more,and there were a large number of autophagosomes in the cells,some epithelial cells were lysed,ischemia In the reperfusion group,part of the mitochondrial crest of the tissue cells disappeared,the volume increased,some cells showed vacuolization,microvilli decreased more,and there were a large number of autophagosomes in the cells,some epithelial cells were lysed,and the vacuole-like changes Compared with the sham operation group,the total rna value of the three oxygen pretreatment group and the ischemia-reperfusion group were larger(P<0.05);the results of Q-PCR analysis by 2-?CT method were three Compared with the oxygen preconditioning group and the ischemia-reperfusion group>1,the total rna value of the ischemic preconditioning group and the ischemic reperfusion group was larger,but P=0.64>0.05,no Statistical significance.It may be related to the length of perfusion time,and the experiment needs to be further improved.Conclusion:Oxygen preconditioning can significantly reduce lung ischemia-reperfusion injury to lung tissue.The mechanism may be that the body is exposed to short and multiple oxidative stress reactions,and the body's anti-oxidative stress system is activated in advance.During the oxidative stress response,the inflammatory response is reduced by removing excess ROS from the body.Oxygen preconditioning can protect the rat from ischemia-reperfusion injury by pre-activating the body's anti-oxidative stress system.Three-oxygen pretreatment can reduce the degree of inflammation and edema of ischemia-reperfused lung tissue.Three oxygen pretreatment can improve hypoxia. |
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