质子泵抑制剂对羟基氯喹治疗的SLE pDCs的影响

Impact of proton pump inhibitor on pDCs in hydroxychloroquine-treated systemic lupus erythematosus patients

目的从体内和体外2个方面探讨质子泵抑制剂(PPI)对羟基氯喹(HCQ)治疗SLE中pDCs影响。方法门诊收集稳定SLE病人27例(HCQ组15例,PPI组12例),流式细胞术测PBMC中pDCs数量和pDCs表面分子表达。磁珠分选得脐带血前体pDCs细胞与HCQ和PPI共培养,并在不同PPI质量浓度时行流式细胞术测pDCs表面分子表达,ELISA检测上清细胞因子IFN-α水平。健康人前体pDCs分别与HCQ和PPI共培养,分为4孔,加荧光探针,激光共聚焦显微镜测pDCs溶酶体pH值变化。结果PPI组与HCQ组相比,外周血中pDCs的数量减少(P<0.05),CD32的表达水平降低(P=0.05);体外培养中,PPI组BDCA-2表达水平低于HCQ组(P<0.05);加入HCQ细胞溶酶体的荧光强度减弱,pH值升高,加入PPI后,其细胞溶酶体荧光强度较HCQ组减弱,pH值进一步升高。结论 PPI和HCQ二者合用降低SLE体内pDCs水平和CD32表达;体外培养中,PPI降低pDCs上BDCA2表达;HCQ和PPI合用可升高pDCs溶酶体内pH值,PPI可能有拮抗HCQ的作用。

To investigate the effects of proton pump inhibitor（PPI） on plasmacytoid dendritic cells（pDCs） of hydroxychloroquine（HCQ）-treated SLE in vitro and in vivo, twenty-seven cases of inactive SLE patients, including15 patients treated with HCQ（HCQ group） and 12 patients treated with HCQ plus PPI（PPI group）, were recruited,and flow cytometry was used to measure the pDCs numbers in PBMC and the expression of pDCs surface molecules.On the other hand, the cord blood precursor pDCs were obtained using beads sorting and co-cultured with HCQ and PPIs in vitro, then the expression of pDCs surface molecules were detected by flow cytometry, while IFN-αlevel in the supernatants was detected using ELISA. The pre-pDCs sorted from healthy donors using magnetic cell separation with BDCA-4 beads were co-cultured respectively with HCQ and PPIs in vitro, in the presence of fluorescent probes, and then the changes of lysosome pH values were determined by laser scanning confocal microscope after 30 min＇s incubation. Compared with HCQ group, PPI could reduced the number of pDCs in PBMC（P0.05） and reduced the expression of CD32 on pDCs（P=0.05） of patients. In vitro, BDCA-2 expression in PPI group was significantly lower than that in HCQ group（P〈0.05）. Furthermore, the fluorescence intensity decreased in HCQ group and further decreased in PPI group accompanied by pH elevation. In conclusion, treatment with PPI plus HCQ results in the decrease of pDCs number and CD32 expression, the reduction of BDCA2 expression on pDCs in vitro,and the elevation of lysosomes pH of pDCs in patients with SLE. Thus PPI may have antagonistic effect on HCQ.