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目的从对巨噬细胞功能调控角度探讨脐带间充质干细胞(umbilical cord mesenchymal stem cells,UCMSC)对狼疮鼠的治疗机制,为临床用于治疗狼疮患者提供理论依据。方法 24周龄B6.MRL-Fas~(lpr)(B6/lpr)狼疮鼠随机分为空白对照组、UCMSC移植治疗组和成纤维样滑膜细胞(fibroblast like synoviocytes,FLS)移植对照组,同周龄雌性C57BL/6鼠作为正常对照。UCMSC和FLS组均给予1×10~6数量尾静脉输注,各组小鼠于28周龄处死。HE检测小鼠肾脏病理,评估疗效。分离纯化小鼠腹腔和肾脏巨噬细胞,无菌条件下取8周龄雌性C57BL/6小鼠脾脏,磁珠分选CD4~+T细胞,在抗CD3/28刺激下,将其以5∶1比例与小鼠巨噬细胞共培养4 d,CFSE法检测CD4~+T细胞增殖。2μm荧光微球与小鼠巨噬细胞共培养2 h,流式细胞术检测巨噬细胞吞噬活性。结果 (1)UCMSC组可显著改善狼疮鼠肾脏病理;(2)UCMSC移植治疗可显著提高狼疮鼠巨噬细胞的免疫调节能力,使其对CD4~+T细胞增殖的抑制能力显著增强;(3)UCMSC组狼疮鼠腹腔巨噬细胞的吞噬功能显著增强。结论 UCMSC移植治疗狼疮鼠可以改善肾脏病理损伤,UCMSC对巨噬细胞免疫调节和吞噬功能的调控是其发挥治疗作用的机制之一。
OBJECTIVE: To investigate the therapeutic mechanism of umbilical cord mesenchymal stem cells (UCMSC) on lupus in rats from the perspective of the functional regulation of macrophages, and to provide a theoretical basis for clinical treatment of lupus. Methods Twenty-four weeks old B6.MRL-Fas ~ (lpr) (B6 / lpr) lupus mice were randomly divided into blank control group, UCMSC transplantation group and fibroblast-like synoviocytes (FLS) transplantation control group Week old female C57BL / 6 mice served as normal control. UCMSC and FLS groups were given 1 × 10 ~ 6 tail vein infusion, mice of each group were sacrificed at 28 weeks of age. HE detection of renal pathology in mice to assess the efficacy. The peritoneal and renal macrophages of mice were isolated and purified. The spleen of 8-week-old female C57BL / 6 mice were collected under sterile conditions. The CD4 ~ + T cells were sorted by magnetic beads. The cells were stimulated with anti-CD3 / 1 ratio was co-cultured with mouse macrophages for 4 days. CFSE assay was used to detect the proliferation of CD4 ~ + T cells. 2μm fluorescent microspheres co-cultured with mouse macrophages for 2 h, macrophage phagocytosis activity was detected by flow cytometry. Results (1) The UCMSC group could significantly improve the renal pathology of the lupus; (2) UCMSC transplantation could significantly enhance the immunomodulatory capacity of macrophages and significantly inhibit the proliferation of CD4 + T cells. (3) ) The phagocytic function of peritoneal macrophages in UCMSC group was significantly increased. Conclusion UCMSC transplantation can improve pathological injury of kidney in mice with lupus, UCMSC plays an important role in the regulation of macrophage immune regulation and phagocytosis.