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目的:探讨趋化因子配体2[chemokine(C-C motif)ligand 2,CCL2]在间充质干细胞(mesenchymal stem cells,MSCs)移植治疗系统性红斑狼疮(systemic lupus erythematosus,SLE)中的作用。方法:采用酶联免疫吸附试验检测SLE患者及正常对照者的血清、尿液及骨髓MSCs培养上清中的CCL2及白细胞介素(interleukin,IL)-17的表达水平,并采用流式细胞术检测外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)中Th17-CD4+IL-17+的百分率及其表面CCR2的表达。将18只雌性MRL/lpr SLE小鼠随机分为SLE小鼠组(n=6)(未治疗组)、脐带MSCs移植联合CCL2中和抗体治疗SLE小鼠组(n=6)及脐带MSCs移植联合同型对照IgG2B治疗SLE小鼠组(n=6),另选6只同周龄雌性C57BL/6小鼠作为健康阴性对照组。采用酶联免疫吸附试验检测各组血清CCL2及IL-17的水平,流式细胞术检测各组PBMCs中CD4+CCR2+IL-17+细胞百分率。结果:SLE患者血清[(839.0±160.5)pg/mL]、尿液[(881.8±162.7)pg/mL]及骨髓MSCs培养上清[(22 935±2 304)pg/mL]的CCL2含量均显著高于正常对照者[(433.0±31.0)pg/mL、(222.7±52.8)pg/mL、(8 394±5 125)pg/mL];SLE患者PBMCs中CD4+CCR2+IL-17+细胞百分率较正常对照者显著升高(P<0.01)。脐带MSCs移植联合CCL2中和抗体治疗SLE小鼠组血清CCL2浓度显著低于未治疗组,PBMCs中CD4+T细胞亚群IL-17、CCR2表达较未治疗组显著降低。结论:MSCs移植治疗SLE的效果可能通过抑制CCL2表达,进而下调患者Th17细胞水平来实现。
Objective: To investigate the role of chemokine (C-C motif) ligand 2 and CCL2 in the treatment of systemic lupus erythematosus (SLE) in mesenchymal stem cells (MSCs) transplantation. Methods: The levels of CCL2 and interleukin (IL) -17 in serum, urine and bone marrow MSCs of SLE patients and normal controls were detected by enzyme-linked immunosorbent assay (ELISA). Flow cytometry The percentage of Th17-CD4 + IL-17 + and its surface CCR2 expression in peripheral blood mononuclear cells (PBMCs) were detected. Eighteen female MRL / lpr SLE mice were randomly divided into SLE group (n = 6) (untreated group), umbilical cord MSCs transplantation combined with CCL2 neutralizing antibody to treat SLE mice (n = 6) and umbilical cord MSCs transplantation The same isotype control IgG2B treatment SLE mice group (n = 6), and the other 6 weeks old female C57BL / 6 mice as a healthy negative control group. The levels of serum CCL2 and IL-17 in each group were detected by enzyme-linked immunosorbent assay (ELISA), and the percentage of CD4 + CCR2 + IL-17 + cells in each group were detected by flow cytometry. Results: The levels of CCL2 in serum of patients with SLE [(839.0 ± 160.5) pg / mL], urine (881.8 ± 162.7 pg / mL) and the culture supernatant of bone marrow MSCs [(22 935 ± 2 304) pg / mL] (433.0 ± 31.0) pg / mL, (222.7 ± 52.8) pg / mL and (8 394 ± 5 125) pg / mL] respectively. CD4 + CCR2 + IL-17 + cells in PBMCs of SLE patients were significantly higher than those in normal controls The percentage was significantly higher than the normal control (P <0.01). The concentration of serum CCL2 in umbilical cord MSCs transplantation combined with CCL2 neutralizing antibody in SLE mice was significantly lower than that in untreated mice. The expression of IL-17 in CD4 + T cells in PBMCs was significantly lower than that in untreated mice. Conclusion: The effect of transplantation of MSCs on SLE may be achieved by inhibiting the expression of CCL2 and then decreasing the level of Th17 cells in patients.