人脐带间充质干细胞对实验性自身免疫性葡萄膜炎的治疗作用研究

Study of human umbilical cord mesenchymal stem cells for treatment of experimental autoimmune uveitis

目的观察体外培养的人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUCMSCs)对实验性自身免疫性葡萄膜炎(experimental autoimmune uveitis,EAU)的治疗作用。方法组织块贴壁法培养HUCMSCs、流式细胞术鉴定。用视网膜光感受器间维生素A类结合蛋白(interphotoreceptor retinoid-binding protein,IRBP)诱导EAU模型。C57BL/6小鼠尾静脉注射HUCMSCs,每日用裂隙灯显微镜对小鼠眼前节的炎症情况进行观察。流式细胞术检测实验小鼠脾脏淋巴细胞CD11c、CD86表型。结果建模后的C57BL/6小鼠于注射IRBP后7~9 d开始发病,12~14 d发病达高峰,16~20 d眼前节炎症自限性减轻,21 d后眼内炎症逐渐消退、自愈。HUCMSCs治疗后,建模小鼠EAU发病时间延长,炎症程度减轻,实验组与模型对照组比较,差异有统计学意义(P<0.05)。建模后的小鼠脾脏CD11c+CD86-细胞数量增加非常明显,经HUCMSCs治疗后第9天CD11c^+CD86^-细胞数量接近治疗前水平。建模后小鼠脾脏中CD86+CD11c+/CD11c比值下降,经HUCMSCs治疗后有所回升,但在第15天时仍未恢复至正常水平。结论 HUCMSCs对IRBP诱导的小鼠EAU有治疗作用,HUCMSCs作用于树突状细胞等抗原呈递细胞,抑制了EAU的进展。

Objective To explore an effective treatment schedules of human umbilical cord mesenchymal stem cells( HUCMSCs) for experimental autoimmune uveitis( EAU) in C57BL/6mice. Methods HUCMSCs were obtained by culturing cesarean section tissues from fetal umbilical cord in vitro by tissues adherent methods. HUCMSCs were identified by flow cytometry. EAU model of C57BL/6 mice were established by using interphotoreceptor retinoid-binding protein( IRBP).HUCMSCs were injected intravenous in C57BL/6 mice. Examine anterior asgment inflammation and hyphema by slit lamp microscope. The expression of CD11 c and CD86 molecular of spleen lymphocytes were analyzed by flow cytometry. Results During the course of IRBP induced EAU in C57BL/6 mice,inflammatory reactions began at 7 to 9 days,peaked at 12 to 14 days and self relieved at 16 to 20 days. The intraocular inflammation disappeared entirely and spontaneous healed after 21 days.The onset time were prolonged and the inflammatory reaction were alleviated in IRBP induced EAU C57BL/6 mice,which had statistical significance( P < 0. 05). The amounts of CD11c+CD86-cells increased obviously in EAU mice and recovered at 9 days after administrated with HUCMSCs. The ratio of CD86+CD11c+/CD11 c decreased in EAU mice and recovered gradually after administrated with HUCMSCs but still not to normal levels at 15 days. Conclusion The treatment method of HUCMSCs was effective for IRBP induced EAU in C57BL/6 mice. The data of CD11c+CD86-cells from flow cytometry supported previous suggestions that HUCMSCs inhibit actions of dendritic cells substantially ameliorate the progression of EAU.