手术应激对儿童外周血淋巴细胞凋亡的影响

The Effect of Apoptosis in Circulating Lymphocytes from Children Undergoing Surgery and General Anesthesia

目的 探讨手术应激后儿童外周血淋巴细胞的凋亡。方法 选择择期手术患儿 16例 ,分别于术前 2 4h、术后 2 4h、术后 72h采集外周血 3.5～ 5ml。用梯度离心法分离出淋巴细胞 ,置于RPMI 16 4 0培养基中 ,培养 15h。然后用 7 AAD染色 ,通过流式细胞仪检测CD4 +、CD8+细胞凋亡。结果 术后 2 4hCD4 +、CD8+淋巴细胞凋亡率 ( % )明显增加 (术前 2 4h分别为 2 .4 4± 0 .4 5和 2 .96± 0 .6 0 ,术后 2 4h为 6 .5 3± 1.96和 5 .13± 1.19) (P <0 .0 5 ) ,术后 72h又恢复至术前水平 ( 3.6 0± 1.0 8和 4 .0 1± 0 .93,P >0 .0 5 )。结论 手术创伤可使儿童外周血CD4 +、CD8+淋巴细胞凋亡增加。

Objective To investigate the apoptosis in children's circulating lymphocytes undergoing surgery and general anesthesia. Methods Sixteen children who underwent elective surgery and general anesthesia were studied.Blood sampling for determination of lymphocytes were performed on the day before surgery (T 0)and at 24 and 72 hours after operation(T 1and T 2)respectively.After isolated,the lymphocytes were maintained in RPMI_1640 about 15 hours.Then,the short_term cultured cells were stained with 7_amino_actinomycin D(7_AAD) to quantify the apoptosis by a FACScan flow cytometer. Results A significantly increased rate of apoptotic CD4 + and CD8 + cells ( P <0.05) was found at the time of T 1 (6.53±1.96 and 5.13±1.19 for CD4 + and CD8 +,respectively). The overall level returned to the baselines ( P >0.05) at 72 hours after surgery.(3.60±1.08 and 4.01±0.93.for CD4 + and CD8 +,respectively). Conclusion Surgical stress is associated with a significant but transient increase in lymphocyte commitment to apoptosis.