The Research Of Oxygen-enriched Perfluorocarbon Emulsion's Protection For The Oleic Acid Rabbit Model Of Acute Respiratory Distressed Syndrome

Public emergency,such as major natural disasters,major mine disaster,tall building fire disaster accidents,frequently occurred,causing large areas of heavy casualties.According to statistics,the first peak of death about traumatic injuries is within 1 h.At this time the number of deaths accounted for 50% of trauma death by severe trauma directly.And the second peak of death is within 2-4 h after injury.This part of the injured is mainly due to cause death because of the shock and hypoxemia.According to reports,acute respiratory distress syndrome is the main cause of death on the battlefield and the leading cause of death in critically ill patients in times of peace.Therefore,hypoxemia is a major cause of cf death in incident,but the existing emergency products have disadvantages such as carrying inconvenience and working slowly,leading to the deadliest in the first place eventually.To develop rapid,efficient and portable emergency products is an important material basis of emergency treatment and protection.This not only has a broad application prospect but also has important social significance.Perfluocarbon has good function of nonpolar gas dissolved,so it can be used as the carrier of oxygen and carbon dioxide.At the same time,dipalmitoylphosphatidylcholine is the main components of the lung surface active substance and can decorate perfluorocarbon to make perfluorocarbon emulsion,providing the effect of improving lung compliance.PFC emulsion has an ability to improve hypoxemia conditions:(1)It is easy to quickly dissolve and release a large amount of oxygen;(2)PFC and PS have ability to improve lung compliance.In addition,there are a lot of the literature of PFC about inhibiting the action of the inflammatory response.So we assume:(1)Atomization inhalation of PFC emulsion could rapidly provides a wide range of oxygen to be aimed at improving hypoxemia.(2)PFC and PS improve lung compliance,achieving the goal of indirectly improving hypoxemia.Part ?Study on the toxicity and reoxygenation effects of the oxygen-enriched perfluorocarbon emulsion on HCT116 cells.Objective To observe the cytotoxicity of oxygen-enriched perfluorocarbon emulsion in vitro and its ability to supply oxygen to hypoxic HCT116 cells.Methods In vitro cultured HCT116 cells were incubated in medium containing perfluorocarbon emulsion of different concentrations(0,0.03,0.06,0.12,0.25,0.5 mg / ml)for 4,8 and 12 h,and the cell proliferation rate was measured by CCK-8 method,so as to analyze the cell toxicity.HCT116 cells were divided into control group(always incubated in normal media and normal oxygen concentration),group at 4 hours of hypoxia and groupat 8 hours of hypoxia,made into hypoxia model with liquid paraffin,then incubated in media containing perfluorocarbon emulsion of different concentrations(0,0.05,0.1,0.2mg/ml)for 2 hours to reoxygenate,CCK-8 method were also used to measure the rate of cell proliferation.After exposing to hypoxia for 12 hours(control group except),the HCT116 cells were added into media containing perfluorocarbon emulsion of different concentrations(0,0.05,0.1,0.2 mg/ml)and reoxygenated for 2 hours,then the DCF fluorescence was detected by fluorescence microscopy,so as to analyze the oxygen capacity of the emulsion.Results On the study of cytotoxicity,a rise in cell proliferation rate were observed to a certain extent(104%,105%,102% and 104%,101%)after HCT116 cells were incubated with perfluorocarbon emulsion of successively concentration gradients for 4 hours;after incubated with perfluorocarbon emulsion of successively concentration gradients for 8hours,a slight decrease(98%,97%)in cell proliferation rate was observed(with concentration of 0.25 or 0.5mg/ml),the rest were increased to some extent;and after incubating for 12 hours,a decrease was observed under the concentration of 0.5mg/ml,but still higher than 90%.Compared with the control group,the Cell proliferation rate was decreased significantly after exposing to hypoxia(88%,76%),and the 8 hours of hypoxia group decreased more significantly than the 4 hours of hypoxia group.After reoxygenating in the media containing perfluorocarbon emulsion,the cell proliferation rate was significantly increased with the gradual increase of the perfluorocarbon emulsion concentration,when compared with the control group,Among them,the cell proliferation rate was increased from 88% to 99% after 4 hours of hypoxia and the difference was statistically significant between 0.1 mg / mL concentration and 0 mg/ml concentration.The cell proliferation rate was increased from 76% to 99% after 8 hours of hypoxia and the difference was statistically significant between 0.2 mg / mL concentration and 0 mg/ml.DCFH-DA probe was used to detect the amount of reactive oxygen species in HCT116 cells after hypoxia and re oxygenation.There was no obvious DCF fluorescence in normal culture group,and the DCF green fluorescence was seen in 12 hours after hypoxia treatment.At the same time,by giving perfluorocarbon emulsion reoxygenation treatment after 12 hours of hypoxia,DCF fluorescence was further enhanced and gradually increased with the increase of perfluorocarbon emulsion concentration,suggesting the increase of intracellular reactive oxygen species content.Conclusions There are oxygen-riched perfluorocarbon emulsion on HCT116 cells without significant cytotoxicity,and oxygen carried by the emulsion can be released for uptake and use by hypoxia HCT116 cell.Part ?The research of oxygen-enriched perfluorocarbon emulsion's protection for the oleic acid rabbit model of acute respiratory distressed syndrome.Objective Discuss the protection of oxygen-enriched perfluorocarbon emulsion for the oleic acid rabbit model of acute respiratory distressed syndrome.Methods Eighteen rabbits after anesthesia were randomly divided into three groups(n=6).BC group was the control group.We only inserted PE-50 polyethylene catheter into the right femoral artery and vein operation without any remaining intervention.OA group was oleic acid group.After successfully modeling,rabbits nebulized saline for15 min.OE group was oleic acid + emulsion group.After successfully modeling,rabbits inhaled oxygen-enriched perfluorocarbon emulsion for 15 min.OA group and OE groups were injected 0.1ml / kg oleic acid from ear vein to establish ARDS model,BC group injected with same saline.Observe and record general condition,respiratory rate,blood oxygen,Smith lung injury score,lung coefficient,lung permeability index,TNF-?,IL-1?,IL-10 concentrations in lung lavage fluid and serum,lung tissue by light microscopy at baseline,molding,0,5,15,30,60,120,240,360 min time after inhalation.Results After observing normal circumstances,we found that respiratory rate to rise,lips cyanosis,restlessness and other symptoms of hypoxia after oleic acid injection in rabbits.In the control group pulmonary lung surface was pale pink and we did not see obvious foam sample liquid in trachea and transverse section of lung.In the oleic acid group we saw hemorrhage and necrosis,hyperemia and edema on the surface.Volume was increased obviously and a lot of pink foamy fluid flowed from transverse section and trachea.In oleic acid + emulsion group we saw visible dotted hemorrhage and necrosis on lung surface and a small pink foamy fluid flowed from transverse section and trachea.When building model successfully,compared with the control group(53.0 ± 11.2times/min),respiratory rate of oleic acid group and oleic acid + emulsion group(108.7 ±24.1 times/min,106.3 ± 30.1 times/min)was significantly increased.The difference was statistically significant(P <0.05).After modeling for 1 hour,compared with the control group(113.5±7.4 mmHg),oleic acid group and oleic acid + emulsion group PO2(58.2±3.2mmHg,60.2±2.5 mmHg)was significantly lower,the difference was statisticallysignificant(P<0.05).At 0min after inhaling perfluorocarbon emulsion,PO2 of oleic acid +emulsion group previous rose(67.2±11.8mmHg),but oleic acid group previous declined(53.3±6.0mmHg).The difference was statistically significant(P =0.035).At 360 min after inhaling perfluorocarbon emulsion,compared with the oleic acid group(66.2 ± 15.2mmHg),PO2 of oleic acid + emulsion group previous rose(92.2 ± 23.1 mmHg).The difference was statistically significant(P =0.044).Compared with the control group(0.6 ±0.4),Smith lung injury score of oleic acid group and oleic acid + emulsion group previous rose(10.7±0.8?9.3±1.0).The difference was statistically significant(P<0.05).Compared with the oleic acid group(10.7±0.8),Smith lung injury score of oleic acid + emulsion group was lower(9.3±1.0).The difference was statistically significant(P=0.026).Compared with the control group(0.5±0.1),lung coefficient of oleic acid group and oleic acid + emulsion group previous rose(1.2±0.2?1.0±0.1).The difference was statistically significant(P<0.05).Compared with the oleic acid group(1.2±0.2),lung coefficient of oleic acid + emulsion group was lower(1.0±0.1).The difference was statistically significant(P=0.033).Compared with the control group(2.9 ± 0.3),lung permeability index of oleic acid group and oleic acid + emulsion group previous rose(3.5±0.4?3.9±0.5).The difference was statistically significant(P < 0.05).At 360 min after inhalation,Compared with the control group(32.7±7.4 pg/ml),TNF-? of oleic acid group and oleic acid + emulsion group previous rose(48.5±13.9 pg/ml?54.9±10.1 pg/ml).The difference was statistically significant(P<0.05).Compared with the control group,TNF-?,IL-1?,IL-10 concentrations in lung lavage fluid of oleic acid group and oleic acid + emulsion group previous rose.The difference was statistically significant(P<0.05).In terms of lung tissue by light microscopy,the control group has normal lung tissue performance.The oleic acid group shows a large pale pink edema fluid and inflammatory cell in the alveoli,a lot of blood vessels in the blood vessels and the alveoli,and some alveolar atelectatic or disappear.After treatment with perfluorocarbon emulsions,the oleic acid + oleic acid group shows that various aspects of lung injury was significantly reduced.Conclusions Oxygen-enriched perfluorocarbon emulsion can increase blood oxygen for the oleic acid rabbit model of ARDS and reduce the lung injury.But inflammatory cytokines of the serum and bronchoalveolar lavage fluid was not reduced.Enrich further study about perfluorocarbon emulsion mechanism of action and the active ingredient.