感染性休克患者外周血T淋巴细胞凋亡和影响因素分析

目的观察感染性休克患者的外周血T淋巴细胞的凋亡情况,并对可能诱导T淋巴细胞凋亡的原因进行探讨。方法采集南京市第一医院ICU自2006年1月至2009年1月收治的32例感染性休克患者的外周血,用Annexin-V法和流式细胞仪测定其中的T淋巴细胞的凋亡情况,同时用酶联免疫法（ELISA）测定血浆中肿瘤坏死因子α（TNF-α）、白细胞介素10（IL-10）、白细胞介素1β（IL-1β）的浓度,用化学发光法检测血浆中的游离皮质醇浓度,将上述的结果和脓毒血症病患者及健康对照（n=22）相比。结果在初始状态下,外周血T淋巴细胞CD4＋亚群在健康对照组、脓毒血症组及感染性休克组分别为（4.41±1.45）%、（7.87±3.82）%及（11.01±4.52）%,感染性休克患者的外周血T淋巴细胞CD4＋亚群的凋亡比例高于脓毒血症患者（P〈0.05）,脓毒血症患者高于健康对照组（P〈0.05）;而初始状态下健康对照组、脓毒血症组及感染性休克组患者外周血T淋巴细胞CD8＋亚群的凋亡为（9.62±8.32）%、（13.09±15.84）%及（11.33±19.62）%,3组比较差异无统计学意义（P〉0.05）。在基础状态下TNF-α的浓度在健康对照组,脓毒血症组及感染性休克组中分别为（16.44±9.55）ng/L、（29.58±13.6）ng/L及（47.99±25.63）ng/L,感染性休克组TNF-α的浓度高于脓毒血症组（P〈0.05）,脓毒血症组高于健康对照组（P〈0.05）,持续72小时;IL-10的浓度在健康对照组,脓毒血症组及感染性休克组中分别为（6.38±7.5）ng/L、（9.67±4.88）ng/L及（15.01±5.36）ng/L,感染性休克组中的IL-10的浓度高于脓毒血症组（P〈0.05）,脓毒血症组高于健康对照组（P〈0.05）,持续72小时;游离皮质醇的浓度在健康对照组,脓毒血症组及感染性休克组中分别为（0.45±0.42）μmol/L、（0.87±0.49）μmol/L及（1.39±0.84）μmol/L,感染性休克组中的游离皮质醇浓度高于脓毒血症组（P〈0.05）,脓毒血症组高于健康对照组（P〈0.05）,持续72小时;IL-1β在健康对照组、脓毒血症组及感染性休克组中分别为（13.12±5.07）ng/L、（25.21±14.76）ng/L及（22.94±22.01）ng/L,感染性休克组和脓毒血症组的浓度高于健康对照组（P〈0.05）,但是感染性休克组和脓毒血症组的浓度差异无统计学意义（P〉0.05）。结论感染性休克患者的外周血T淋巴细胞中CD4＋亚群可发生明显的凋亡,TNF-α、IL-10及皮质醇可能参与了诱导CD4＋亚群的凋亡。

Investigation of circulating T lymphocyte apoptosis and influencial factors in septic shock patients

Objective To investigate circulating T lymphocyte apoptosis in septic shock and the factors that may induce T lymphocyte apoptosis. Methods Peripheral venous blood samples of 32 patients with septic shock from January 2006 to January 2009 in Intensive Care Unit of the Nanjing First Hospital were obtained to investigate the circulating T lymphocyte apoptosis by Annexin V and flow cytometry.In the meantime,the plasma concentration of tumor necrosis-α（TNF-α）,interleukin-10（IL-10）,interleukin-1β（IL-1β） were measured with ELISA and free cortisol was measured with chemiluminescence.All these results were compared between sepsis patients and healthy volunteers（n=22）. Results In baseline,the percentages of annexin Ⅴ-positive CD4＋T lymphocytes in healthy volunteers,sepsis patients and septic shock were（4.41±1.45）%,（7.87±3.82）% and（11.01±4.52）%.And the percentage of annexin Ⅴ-positive CD4＋T lymphocytes was more in septic shock than that in sepsis patients（P0.05）,and the latter was more than healthy volunteers（P0.05）.In the initial setting,the percentages of annexin Ⅴ-positive CD8＋T lymphocytes in healthy volunteers,sepsis patients and septic shock were（9.62±8.32）%,（13.09±15.84）% and（11.33±19.62）%,respectively,the conditions were no statistic differences among three groups（P0.05）.The plasma concentrations of TNF-α in healthy volunteers,sepsis patients and septic shock in baseline were（16.44±9.55） ng/L,（29.58±13.6） ng/L and（47.99±25.63） ng/L,respectively.And,TNF-α was higher in septic shock than that in septic patients（P0.05）,TNF-α was higher in septic patients than that in healthy volunteers（P0.05）.This condition remained for 72 hours.The plasma concentrations of IL-10 in healthy volunteers,sepsis patients and septic shock in baseline were（6.38±7.5） ng/L,（9.67±4.88） ng/L and（15.01±5.36） ng/L,respectively.IL-10 was higher in septic shock than that in sepsis patients（P0.05）,IL-10 was higher than that in healthy volunteers（P0.05）.This remained for 72 hours,too.The plasma concentrations of free cortisol in healthy volunteers,sepsis patients and septic shock in baseline were（0.45±0.42） μmol/L,（0.87±0.49） μmol/L,（1.39±0.84） μmol/L.Free cortisol was higher in septic shock than that in sepsis patients（P0.05）.And the latter was higher than that in healthy volunteers（P0.05）.This condition also remained for 72 hours.The plasma concentrations of IL-1β in healthy volunteers,sepsis patients and septic shock in baseline were（13.12±5.07） ng/L,（25.21±14.76） ng/L and（22.94±22.01） ng/L,respectively.IL-1β was higher in sepsis patients and septic shock than that in healthy volunteers（P0.05）.However,there was no statistic difference between sepsis patients and septic shock（P0.05）. Conclusion Circulating CD4＋ T lymphocyte apoptosis was higher in septic shock.TNF-α,IL-10 and cortisol may be involved in CD4＋ T lymphocyte apoptosis.