| Part1Preparation of rat amniotic fluid embolism model,identification and peripheral hemodynamic changeObjective: To prepare a rat amniotic fluid embolism model and identify,observe theperipheral hemodynamic changes.Methods: Thirty SD pregnant rats were divided into three groups randomly;normalsaline group(control group),amniotic fluid group(the experimental group1),meconium-stained amniotic supernatant fluid group(the experimental group2). Afteranesthesia,made a subtotal hysterectomy for rat and closed abdomen.Separated leftcommon carotid artery,made a left common carotid arterial cannula and switched thethree-way lube the link the two channels physiological recorder. Respectively injection ofsaline,amniotic liquid, meconium-stained amniotic supernatant fluid through the abdominalvein,at60min killed the animals with10％KCL.Taken rat lung fixed,paraffin-embedded,Sections were stained with HE and APM to find amniotic fluidcomposition. Immunohistochemistry detected pulmonary intravascular CK10expression,and monitor Hemodynamic indexes. Peripheral hemodynamic data were using repeatedmeasures analysis of variance and LSD method, P <0.05considered statisticallysignificant.Results: Successfully established rat amniotic fluid embolism model,the control group allwere Survival, the experimental group1rats observed tachypnea,dysphoria,cyanosis, nosewet-cold,aconuresis,no death within60min, rats symptoms were severe than the theexperimental group2,there one rat died within60min. The experimental group showedpulmonary edema in different degrees with HE staining,large of neutrophils invasion, thelung small blood vessels observed keratinized10stratified squamous epithelium andvisible components of amniotic fluid,the extent of damage was heavier in the experimentalgroup2, the control group have not changed significantly. View keratinizing squamousepithelium were pink in APM,mucus were blue,Neutrophil cytoplasmic light blue orcolorless and red nuclear,control group was not observed. Immunohistochemistry of CK10in the the experimental group,observed brown filamentous keratinized squamous epithelium positive substance in pulmonary vascular, the control group was not observed.There were transienthemodynamic changes after normal sodium solution infusion, but hadno significant (P>0.05). Compared with the group one,the other two groups Systolic bloodpressure, Diastolic blood pressure and Mean arterial pressure decreased significantly (P<0.05),10min after injection of amniotic fluid was most obvious,30min to60min baselineremained low state,one rat MAP of the third group decreased to20mmHg,Finallydeveloped into heart failure,The second and third group were no significant differencebetween groups (P>0.05).Three groups showed no statistically significant in heart rate (P>0.05).Conclusion: Amniotic fluid embolism in rats as animal model was more appropriate,economic, convenient operation, it could be recommended in the amniotic fluid embolismlaboratory studies.HE staining,APM staining and Immunohistochemical stainingcytokeratin-10could assist the diagnosis of amniotic fluid embolism, which was sensitiveand objective method. when AFE occurs,the first signs might be peripheral hemodynamicindicators. Part2The experimental study of phospholipase A2and PAF in RatAFE pathogenesisObjective: To research sPLA2and PAF in rat amniotic fluid embolism pathogenesis,observed cPLA2expression in pulmonary, providing theoretical basis for the etiology andpathogenesis of amniotic fluid embolism.Methods: After the first part of the preparation of an animal model of amniotic fluidembolism is completed, drew1ml blood from left common carotid artery atpro-experiment and post-experiment60min respectively. Immunohistochemistry detectedpulmonary cPLA2expression. All data were analyzed by using SPSS19.0statisticalsoftware and were expressed as median standard error. T test,covariance and correlation®ression were used to analyze the data．Results:1、Three groups pro-experiment and post-experiment sPLA2concentration: the controlgroup were747.62±74.01ng/L and769.91±95.42ng/L;the experimental group1were733.20±76.50ng/L and1007.72±107.04ng/L; the experimental group2were721.96±113.21ng/L and1189.87±264.82ng/L.2、Three groups pro-experiment and post-experiment PAF concentration: the controlgroup were28.94±4.34ug/L and30.16±4.95ug/L;the experimental group1were30.45±8.10ug/L and38.93±9.12ug/L;the experimental group2were28.37±5.79ug/Land43.60±9.29ug/L.3、sPLA2and PAF of Serum concentration correlation: pro-experiment sPLA2and PAFhave no correlation (P=0.834,R=0.045);post-experiment sPLA2and PAF werepositively correlated (P=0.006,R=0.548)4、sPLA2and PAF concentration of Amniotic fluid and meconium in rats: sPLA2concentration in Amniotic fluid is about626.42ng/L, in1%to7%concentration ofmeconium-stained amniotic fluid is about999.24ng/L; PAF concentration inAmniotic fluid is about29ug/L, in1%to7%concentration of meconium-stainedamniotic fluid is about35.56ug/L5、Immunohistochemistry of cPLA2in the experimental group,over expressed in bronchialepithelial cells and neutrophils,macrophages or other inflammatory cells cytoplasmicbrown substances,NS group were slightly expressed. Conclusion:The experimental group post-experimental sPLA2and PAF were increased, Indicates thepresence of something in amniotic fluid and meconium, which could stimulate pregnantbody sPLA2and PAF increased.We detectde a certain amount of sPLA2and PAF in theamniotic fluid and meconium-stained amniotic supernatant fluid, both were differences,ithigher levels of in the meconium. Immunohistochemical of cPLA2expressed inbronchiolar epithelium and inflammatory cells, suggest AF could stimulate bronchiolarepithelium and inflammatory cells to secrete cPLA2. |
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