家兔烫伤早期中性粒细胞凋亡变化

Changes of P MN apoptosis in rabbits after burn

目的 通过检测烧伤早期中性粒细胞 (PMN)凋亡 ,观察烧伤血清和创面水肿液对PMN体外自发凋亡的影响。 方法 应用全血荧光染色观察 3 0 %家兔Ⅲ度烫伤早期PMN凋亡 ;应用流式细胞分析等方法观察血清和痂下组织液培养PMN的体外凋亡。 结果 伤后 4、8、12、2 4h循环PMN凋亡减少 ,约为伤前的 1/ 6,且 4个时相间差异无显著意义 ;烧伤血清和痂下组织液能抑制PMN凋亡 ,使PMN凋亡率下降 ,DNA断裂百分率降低 ,DNA凝胶电泳不出现“梯状”带 ;而正常血清刺激烧伤家兔PMN时不出现凋亡抑制。 结论 烧伤早期体内PMN凋亡延迟 ;烧伤创面水肿液和烧伤早期血清能抑制PMN凋亡。

Objectives TostudythedynamicchangesofapoptosisofPMNfromrabbitsintheearlystage aftersevereburninvivoandinvestigatetheeffectofburnserumandwoundexudateonPMNapotosisinvitro . Methods 12rabbitswererandomlydividedinto 2groups:controlgroup ( 3 7℃ )andburngroup ( 95℃ ,15sec, Ⅲ° 3 0 %TBSA) .Wholebloodfromtheburngroupat0 ,4 ,8,12 ,2 4hpostburnwasstainedwithacridineorange/ ethidiumbromide (AO +EB) ,andobservedunderUVmicroscopy .PMNswereisolatedfrombothgroupsandcul turedfor2 4h ( 3 7℃ ,5 %CO2 )withnormalserum (NS) ,burnserum (BS) ,andsubeschartissuefluid (STF) respectively.MorphologicalassessmentandquantitationofapoptosiswereperfromedwithAO +EBstainandflow cytometricanalysis. Results ThenumberofapoptoticPMNinvivodecreasedintheearlystagepostburn .Inthe BS +PMNgroupandSTF +PMNgroup ,thepercentageofapoptoticPMNreduced (vsNS +PMNgroup ,P < 0 0 5 )andthespecificDNA“ladder”patternforapoptoticcellswasnotdetectedbyagarosegelelectrophoresis.We observedthatbothburnserumandSTFcouldsimilarlysuppressedPMNapoptosiswhiletherewasnosignificantdif ferencebetweenNS +normalPMNgroupandNS +burnPMNgroup . Conclusions Apoptoisisinhibitedin PMNfromrabbitspostburnintheearlystage .BothburnserumandSTFfromrabbitspostburncaninhibitPMN apoptosisinvitro .