创伤患者外周血单核细胞自噬变化的研究

Autophagy of peripheral monocytes following trauma:study on 51 cases

目的研究创伤患者外周血单核细胞的自噬变化。方法①根据损伤严重度评分(injury severity score,ISS)或简明损伤定级标准(abbreviated injury scale,AIS)将创伤患者分为轻度创伤组、中度创伤组、严重创伤组,同时设立健康对照组,分别于创伤后24h、3、5、10d采集外周血,采用密度梯度离心结合贴壁分离法分离创伤患者外周血单核细胞,应用激光共聚焦显微镜和透射电镜观察细胞自噬现象。②将分离得到的单核细胞进行MDC染色并裂解,应用荧光分光光度计(激发光波长380nm、发射光波长525nm)测定荧光强度进行定量分析。③于创伤后24h、3、5d检测患者血糖水平,分析创伤后自噬与血糖的相关性。结果与健康对照组相比,创伤患者外周血单核细胞自噬水平明显升高(P<0.01),创伤后外周血单核细胞自噬水平与创伤程度成正比,与创伤后血糖呈正相关(轻度创伤组:r=0.962,中度创伤组:r=0.928,重度创伤组:r=0.973,P<0.05)。结论创伤后通过调节单核细胞自噬水平,使机体免疫功能得到恢复,从而改善预后。

Objective To investigate the change of monocyte autophagy in patients after trauma. Methods （1）According to Injury Severity Score （ISS） or Abbreviated Injury Scale （AIS）, we divided our 51 subjected trauma patients into 3 groups, the minor injury group （n = 21 ）, the moderate trauma group （n = 19） and the severe trauma group （ n = 11 ）. Another 14 healthy volunteers served as control. We collected the peripheral blood in 24 h, 3,5 and 10 d after trauma, then harvested the monocytes by density gradient centrifugation with adherence separation. Then we observed the autophagy phenomenon by using laser confocal microscopy and transmission electron microscopy; （2） After the isolated monocytes were dyed with MDC and lysed, they were detected and quantified for fluorescent intensity by fluorospectrophotometer （excitation wavelength 380 nm and emission wavelength 525 nm） ; （3）Blood sugar level of all patients were measured in 24 h, 3 and 5 d respectively after trauma, and the correlation between autophagy and blood sugar was analyzed. Results Compared with control group, more autopbagy in monocytes was observed and quantified in trauma patients than in health volunteers （P 〈 0.01 ）. Conclusion Autophagy in monocytes is associated with the severity of trauma, and has a positive correlation with blood sugar.