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目的探讨白细胞介素-17(IL-17)对结核病患者外周血中性粒细胞(PMN)凋亡的影响以及可能涉及的信号通路。方法取结核病患者和健康人外周血PMN培养0~24 h,annexinⅤ染色后流式细胞术检测PMN凋亡率;或加不同浓度IL-17刺激培养24 h再检测PMN凋亡率;或用MAPK特异性抑制剂U0126预处理30 min后,再加IL-17培养24 h后检测PMN的凋亡。结果结核病患者和正常人PMN凋亡率均随培养时间延长而增加(P<0.05),结核病患者PMN在0、6、12、24 h时的annexinⅤ+细胞分别为(4.49±1.39)%、(21.89±2.90)%、(39.96±4.15)%、(68.35±7.01)%,均分别高于正常人组的(2.65±0.75)%、(11.00±1.72)%、(25.84±3.90)%,(45.59±4.10)%(P<0.05)。而经IL-17刺激后,结核病患者与正常人PMN的凋亡率,在IL-17 0.5μg/L组,分别为(59.81±7.19)%和(34.65±4.79)%;在IL-17 5μg/L组,分别为(51.62±6.91)%和(29.04±3.62)%;均低于未加IL-17对照组(68.35±7.01)%和(45.59±4.10)%(P<0.05);而在IL-17 50μg/L组,凋亡率则分别达到(76.04±5.59)%和(53.24±4.62)%,明显高于对照组(P<0.05)。预先用U0126处理,可大部分阻断IL-17对结核病患者和正常人PMN凋亡的抑制作用。结论结核病患者和正常人PMN凋亡率均随培养时间延长而增加,且结核病患者PMN凋亡率高于正常人。IL-17较低浓度延迟凋亡,较高浓度促进凋亡。IL-17抑制PMN凋亡的作用和ERK途径有关。
Objective To investigate the effect of interleukin-17 (IL-17) on the apoptosis of peripheral blood neutrophil (PMN) in patients with tuberculosis and the possible signaling pathways. Methods Peripheral blood PMN of TB patients and healthy individuals were cultured for 0-24 h. The apoptotic rate of PMN was detected by annexinⅤ staining or PMN apoptosis rate after 24 h incubation with different concentrations of IL-17. The specific inhibitor U0126 pretreatment 30 min, plus IL-17 cultured 24 h after the detection of PMN apoptosis. Results The apoptotic rates of PMN in tuberculosis patients and normal individuals increased with the prolongation of culture time (P <0.05). The percentages of annexinⅤ + cells in PMN at 0, 6, 12 and 24 hours were (4.49 ± 1.39)%, 21.89 ± 2.90%, 39.96 ± 4.15% and 68.35 ± 7.01%, respectively, which were significantly higher than those in normal controls (2.65 ± 0.75%, 11.00 ± 1.72%, 25.84 ± 3.90%, 45.59 ± 4.10)% (P <0.05). However, the apoptotic rate of PMN in tuberculosis patients and normal subjects was (59.81 ± 7.19)% and (34.65 ± 4.79)% in IL-17 0.5 μg / L group after IL-17 stimulation, / L group were (51.62 ± 6.91)% and (29.04 ± 3.62)%, respectively, which were lower than those in the control group without IL-17 (68.35 ± 7.01% vs 45.59 ± 4.10%, P <0.05) The apoptosis rate in IL-17 50μg / L group was (76.04 ± 5.59)% and (53.24 ± 4.62)%, which was significantly higher than that in control group (P <0.05). Pretreatment with U0126 can largely block the inhibitory effect of IL-17 on PMN apoptosis in both TB patients and normal individuals. Conclusion The apoptosis rate of PMN in tuberculosis patients and normal individuals both increases with the prolongation of culture time, and the apoptosis rate of PMN in tuberculosis patients is higher than that in normal people. Lower concentrations of IL-17 delay apoptosis, and higher concentrations promote apoptosis. The role of IL-17 in inhibiting PMN apoptosis is related to the ERK pathway.