目的:探讨Ⅱ型固有淋巴细胞(ILC2)在慢性肾衰发生发展中的作用及其可能机制。方法:选择中山大学附属第一医院2016年3月~2016年12月入院的慢性肾功能衰竭患者36例,选取同期健康体检者32例为对照。流式细胞术(FCM)检测外周血单个核细胞(PBMC)中ILC2的比例;ELISA技术检测血浆中白细胞介素(IL)-13的浓度;提取慢性肾衰患者及健康对照者PBMC后分别分为3组(对照组、细胞因子刺激组、干预组)进行体外培养3 d后,ELISA法测定上清液中IL-13浓度;Western blot法对健康对照者PBMC在刺激前及刺激后15 min、30 min、1 h、2 h的转录活化因子6(STAT6)的磷酸化水平进行检测。结果:慢性肾功能衰竭患者PBMC中ILC2比例及血浆IL-13浓度均较健康人高(P〈0.05);体外培养上清液中,慢性肾功能衰竭患者的3个亚组中IL-13浓度均较健康人高(P〈0.05),且2类人群中均呈现出细胞因子刺激组较对照组升高,干预组较细胞因子刺激组降低的现象;Western blot结果显示p-STAT6的蛋白水平随时间的延长逐渐增高。结论:慢性肾衰患者外周血中ILC2的比例升高,同时ILC2内STAT6活化并分泌大量IL-13,介导Th2细胞的极化而调节免疫。
AIM: To explore the effect and possible mechanism of type 2 innate lymphoid cell (ILC2) on the development of chronic renal failure ( CRF) . METHODS: The patients with chronic renal failure (n=36) in the Fist Af-filiated Hospital of Sun Yat-sen University from March 2016 to December 2016 were selected, and 32 healthy persons in the same period were enrolled in the study for control. The proportion of ILC2 in the PBMC of CRF patients and healthy con-trols was detected by flow cytometry, IL-13 concentration in the plasma was measured by ELISA. The isolated PBMCs from the patients and healthy persons were divided into 3 groups ( control group, cytokine group, intervention group) and cul-tured in vitro for 3 days, respespestively, then IL-13 concentration was measured by ELISA. The protein levels of phospho- rylated signal transducers and activators of transcription 6 ( p-STAT6) in the PBMC of healthy controls before stimulation and after stimulation for 15 min, 30 min, 1 h, 2 h were determined by Western blot. RESULTS: The proportion of ILC2 in the PBMC and the plasma IL-13 concentration of CRF patients was higher than that in the healthy controls (P 〈0. 0 5 ) . In the culture supernatant in vitro^ IL-13 concentration in the 3 subgroups of CRF patients (control group, cytokine group, intervention group) were all higher than that in the healthy controls (P 〈0. 05) , both the 2 groups showed a trend that the active IL-13 concentration in cytokine group was higher than that in control group, and that in intervention group was lower than that in cytokine group. The protein levels of p-STAT6 in cytokine stimulated-PBMC with a time dependent manner. CONCLUSION: The percentage of ILC2 in the PBMC is elevated in CRF patients. Furthermore, the ILC2 secret large a-mount of IL-13 to mediate the polarization of Th2 cells to regulate immunity through activating p-STAT6.