摘要
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annexin Ⅴ＋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 0.5 μg/L组, 分别为（59.81±7.19）%和（34.65 ±4.79）%; 在IL-17 5 μg/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 50 μg/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 （IL）-17 on the apoptosis of neutrophils （PMNs） from peripheral blood of patients with pulmonary tuberculosis （TB） and explore the possible involved signaling pathway. Methods The fresh isolated PMNs from peripheral blood of TB patients and healthy adults were cultured for 0 - 24 hours, and then stained with annexin V. The flow cytometry was used to measure cell apoptosis of PMNs. The fresh isolated PMNs were cultured with different concentrations of IL-17 for 24 hours, with or without pretreatment of MAPK inhibitor U0126 for 30 minutes, the apoptosis of PMNs was detected by flow cytometry. Results The apoptosis of PMNs in both TB patients and healthy adults increased with the culture time going on （P 〈 0.05）. The spontaneous apoptotic （annexin V ＋） rates of PMNs at 0, 6, 12 and 24 hours of culture time in TB patients were （4.49 ± 1.39）%, （21.89 ±2.90）%, （39.96 ±4.15）%, and （68.3.5 ±7.01）%, respectively, which were significantly higher than the corresponding ones in normal healthy groups, （2.65 ± 0.75 ） %, （ 11.00 ± 1.72） %, （25.84 ± 3.90）%, and （45.59 ± 4. 10）%, respectively （P 〈 0.05）. After cultured with I L-17 for 24 hours, the apoptotic rates of PMNs in TB patients and healthy adults were respectively （59.81 ± 7.19）% and （34.65 ±4.79）% in the group of IL-17 at 0.5 μg/L, and （51.62 ±6.91 ）% and （29.04 ±3.62）% in the group of IL-17 at 5 μg/L; they were significantly lower than those in control groups without IL-17, （68.35 ± 7.01 ） % and （45.59 ± 4.10） %, respectively, （ P 〈 0.05）. However, in the group of IL-17 at 50 μg/L, the apoptosis rates of PMNs in TB patients and healthy adults were （76.04 ±5.59）% and （53.24 ± 4.62） %, respectively, which were obviously higher than those of the control groups without IL-17 （P 〈 0.0.5 ）. Pretreatment of PMNs with U0126 mostly inhibited the anti-apoptosis effect of IL-17 at 5 μg/L on PMNs of TB patients and normal subjects.Conclusion The apoptosis of PMNs in both TB patients and healthy adults increases along with the culture time, and the apoptosis rate of PMNs in TB patients is higher than that of healthy adults. The lower concentrations of IL-17 can delay, but the higher concentration of 11-17 can accelerate the apoptosis of PMNs in TB patients. The inhibitory role of IL-17 in apoptosis of PMNs may be involved in the ERK pathway.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2014年第8期833-836,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
高等学校优秀青年人才基金(2012SQRL098)
安徽省高等学校省级自然科学研究(kj2012z52
kj2011z253)
安徽省高校省级自然科学研究(KJ2013B142)
国家科技重大专项基金课题(2008ZX10003011)
