烧伤病人早期血清和创面水肿液对中性粒细胞凋亡的影响

Effect of serum and wound exudates on PMN apoptosis in burned patients

目的 探讨烧伤病人早期血清、水泡液和痂下组织液对中性粒细胞 (PMN)凋亡的影响。 方法 采用伤后 3d内重度以上烧伤患者血清、2 4h内水泡液以及 1周内Ⅲ度创面痂下组织液体外培养正常PMN ,以正常血清作对照 ,应用流式细胞分析等方法观察PMN凋亡。 结果 烧伤病人早期血清、水泡液和痂下液能抑制PMN凋亡 ,使PMN凋亡百分率下降、DNA断裂百分率降低 ,DAN凝胶电泳不出现细胞凋亡特有的“梯形带” ;且痂下液与烧伤病人早期血清对PMN凋亡的抑制程度几乎相同 ,而水泡液的抑制作用稍弱于前两者。 结论 烧伤病人早期血清。

Objective To study in vitro the effect of burn serum and wound exudates on PMN apoptosis in burned patients. Methods The burn serum was obtained from patients within 3 days postburn. The blister fluid was collected from intact blisters in burn wound within 24 hours postburn, and subeschar tissue fluid underneath Ⅲdegree burn wound during escharectomy. PMNs were isolated from healthy human donors and cultured for 24 hours at 37 ℃ and 5%CO 2 with normal serum (NS), burn serum (BS),burn blister fluid (BBF) and subeschar tissue fluid (STF), respectively.Morphological assessment and quantification of PMN apoptosis were performed with AO+EB stain and flow cytometric analysis. Results The number of apoptotic PMN and DNA fragmentation of PMN significantly decreased ( P<0.05) when PMN were cultured with BS, BBF, and STF when compared with that with NS. In addition, the specific DNA 'ladder' pattern for apoptotic cells couldn't be found on the agarose gel electrophoresis in this case. However, BBF exhibited weaker inhibiting effect than BS and STF on PMN apoptosis. Conclusion PMN apoptosis couldbe inhibited in vitro by BS,STF and BBF from burned patients.