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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 study the changes of autophagy in peripheral blood mononuclear cells in traumatic patients. Methods ①According to injury severity score (ISS) or abbreviated injury scale (AIS), the trauma patients were divided into mild trauma group, moderate trauma group and severe trauma group, meanwhile healthy control group The peripheral blood mononuclear cells (PBMCs) were collected at 24h, 3,5 and 10d after trauma. Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation and adherent separation. The autophagy was observed by laser confocal microscopy and transmission electron microscopy. ② The isolated mononuclear cells were stained with MDC and lysed. The fluorescence intensity was measured by fluorescence spectrophotometer (excitation light wavelength 380 nm, emission light wavelength 525 nm) for quantitative analysis. ③ In 24h, 3,5d post-trauma patients blood glucose levels, analysis of post-traumatic autophagy and blood glucose correlation. Results Compared with healthy controls, the level of autophagy in peripheral blood mononuclear cells was significantly increased in trauma patients (P <0.01). The level of autophagy in peripheral blood mononuclear cells after trauma was proportional to the degree of trauma and positively correlated with posttraumatic blood glucose Mild trauma group: r = 0.962, moderate trauma group: r = 0.928, severe trauma group: r = 0.973, P <0.05). Conclusion After the trauma, the immune function of the body is restored by regulating the level of autophagy in monocytes to improve the prognosis.