低剂量氢化可的松对感染性休克患者外周血胸腺依赖性淋巴细胞凋亡的影响

Effects of low dose of hydrocortisone on circulating thymus-dependent lymphocyte apoptosis in patients with septic shock

目的探讨低剂量氢化可的松治疗感染性休克时对外周血胸腺依赖性淋巴细胞（T淋巴细胞）凋亡的影响。方法前瞻性地将南京市第一医院ICU2006年1月至2009年1月收治的57例感染性休克患者，按照随机（随机数字法）对照的原则分为氢化可的松治疗组和对照组，同时选取20例健康志愿者及18例脓毒血症患者进行对比，采取入选的健康志愿者和患者发病的0h，24h，48h，72h，168h的外周血，用AnnexinV法和流式细胞仪，动态测定T淋巴细胞亚群的凋亡情况。组间两两比较时采用LSD—t检验法。结果在初始状态下，感染性休克患者的外周血T淋巴细胞CD4^＋亚群的凋亡为（11．01±4．52）％，健康对照组为（4．41±1．45）％，脓毒血症患者为（7．87±3．82）％；感染性休克患者的外周血T淋巴细胞CD4^＋亚群的凋亡多于健康对照组（P〈0．05）及脓毒血症患者（P〈0．05）；而初始状态下感染性休克患者外周血T淋巴细胞CD8^＋亚群的凋亡为（11．33±19．62）％，健康对照组（9．62±8．32）％，脓毒血症患者（13．09±15．84）％，三组比较差异无统计学意义（P〉0．05）。感染性休克对照组患者24h，48h，72h外周血CD4^＋亚群的凋亡分别为（13．51±6．85）％，（19．39±6．63）％，（15．33±6．21）％，治疗组24h，48h，72h外周血CD4^＋亚群的凋亡分别为（17．4±7．21）％，（22．61±5．64）％，（25．73±6．91）％，治疗组的CD4^＋亚群的凋亡比例明显多于对照组（P〈0．05）；感染性休克对照组患者24h，48h，72h外周血CD8^＋亚群的凋亡分别为（11．49±11．73）％，（12．74±10．39）％，（13．28±16．6）％，治疗组24h，48h，72h外周血CD8^＋亚群的凋亡分别为（9．49±8．9）％，（15．32±18．17）％，（13．68±16．84）％，两组比较差异无统计学意义（P〉0．05）；发病168h后，感染性休克对照组和治疗组外周血CD4^＋亚群的凋亡分别为（5．64±4．58）％vs．（6．79±6．80）％，两组比较差异无统计学意义（P〉0．05），发病168h感染性休克对照组和治疗组外周血CD8^＋亚群的凋亡分别为（12．72±19．69）％vs．（13．88±13．28）％，两组比较差异也无统计学意义（P〉0．05）。结论采用氢化可的松治疗感染性休克时，CD4^＋淋巴细胞凋亡明显增多，而CD8^＋亚群不受影响。

Objective To investigate the effects of low-dose of hydrocortisone on circulating thymus-dependent lymphocyte （T lymphocyte） apoptosis in patients with septic shock. Method Fifty-seven patients with septic shock admitted into ICU from January 2006 to January 2009 were prospectively randomized （random number） to treatment group and control group.Another 20 healthy volunteers and 18 patients with sepsis alone were included as external control groups. The patients of treatment group and control group were treated with low-dose of hydrocortisone and placebo, respectively. Samples of peripheral blood were taken from healthy volunteers and patients 0 hr, 24 hrs,48 hrs,72 his and 168 hrs after onset of the disease to determine the circulating T lymphocyte apaptosis by using the assays of Annexin V and flow cytometry. Least significant difference t -test was used for multiple comparisons. Results The percentage of Annexin V-positive CD4^ ＋ T lymphocytes in the primary stage was （ 11.01± 4.52） % in septic shock patients, （4.41 ± 1.45）% in bealthy volunteers, and （7.87± 3.82）% in patients with sepsis alone. And in the initial setting, the percentage of Annexin V-positive CD4^＋ T lymphocytes in the septic shock patients was higher than that in healthy volunteers （ P 〈 0.05） and in patients with sepsis alone （ P 〈 0.05）. The percentage of Annexin V-positive CD8^＋ T lymphocytes at the beginning was （ 11.33 ± 19.62） % in septic shock patients, （9.62± 8.32）% in healthy volunteers, and （13.09 ±15.84）% in patients with sepsis alone （P 〉 0.05 between three groups）. The percentages of Annexin V-positive CD4 ^＋ T lymphocytes in control group after 24 hrs, 48 hrs and 72 hrs were （ 13.51 ± 6.85） %, （ 19.39 ±6.63） % and （ 15.33 ± 6.21 ） %, respoctively. And the percentages of Annexin V-positive CD4^＋ T lymphocytes in treatment patients after 24 hrs, 48 hrs and 72 hrs were （17.4± 7.21）%, （22.61 ±5.64）%, and （25.73 ±6.91） %, respectively. The percentage of Annexiu V-positive CD4^＋ T lymphocytes in septic shock patients was higher than that in control groups （P 〈 0.05）. The percentages of Annexin V-positive CD8^＋ T 1ymphocytes in control group after 24 hrs, 48 hrs and 72 hrs were （11.49±11.73） %, （ 12.74 ±10.39） % and （ 13.28 ± 16.6） %, respectively, and in the treatment group, those were （9.49 ± 8.9） %, （ 15.32 ± 18.17） % and （ 13.68 ± 16.84） %, respectively （ P 〉 0.05 between two groups）. In the meantime, the percentages of Annexin V-positive CD8^＋ T lymphocytes in control group and in treatment group were （ 12.72 ± 19.69） % and （ 13.88 ± 13.28） %, respectively （ P 〉 0.05）. Conclusions Low-dose of hydrocortisone could induce CD4 ^＋ T lymphocyte apoptosis and has no effects on CD8^ ＋ T lymphocyte apoptosis when it is used to treat septic shock.