骨髓间充质干细胞对兔实验性自体免疫性葡萄膜炎的影响

目的研究骨髓间充质干细胞(BMSCs)对兔实验性自体免疫性葡萄膜炎(EAU)的治疗作用及其发挥作用的可能机制。方法从5只新西兰大白兔股骨骨髓中分离、培养BMSCs并进行传代,第3⁴代细胞用于实验。将24只新西兰大白兔采用随机数字表法随机分为正常对照组、模型对照组和BMSCs干预组,每组8只。正常对照组不作任何处理,模型对照组和BMSCs干预组建立EAU模型。造模成功后将0.1 ml细胞数为2×106/ml的BMSCs悬液经双眼玻璃体腔注射给BMSCs干预组。模型对照组经双眼玻璃体腔注射0.1 ml磷酸盐缓冲液。比较3组新西兰大白兔的眼部表现、组织病理学分级和血清/房水中肿瘤坏死因子-α(TNF-α)水平。结果培养的细胞呈梭形生长,经流式细胞仪检测:CD105(+),CD34和CD45(-)。正常对照组眼前节、眼底表现、B超及苏木素-伊红染色结果未见异常。BMSCs干预组眼前节炎症反应评分及组织病理学分级均低于模型对照组,差异均有统计学意义(P<0.05)。模型对照组血清/房水中TNF-α水平与正常对照组相比升高,差异有统计学意义(P<0.05)。BMSCs干预组血清/房水中TNF-α水平与模型对照组相比下降,差异有统计学意义(P<0.05)。结论 BMSCs对兔EAU有治疗作用,其机制可能是通过降低血清/房水中炎性因子TNF-α的水平。

Effects of bone marrow mesenchymal stem cells on experimental autoimmune uveitis in rabbits

Objective To investigate the therapeutic effects and possible mechanisms that bone marrow mes-enchymal stem cells （BMSCs） impact on experimental autoimmune uveitis （EAU） in rabbits. Methods BMSCs were isolated from the femoral bone marrow of five New Zealand white rabbits for incubation and passage for three and four generations. Twenty-four New Zealand white rabbits were allocated to normal control group （n=8）, model control group （n=8） and BMSCs intervention group （n=8） with a randomized digit table. In normal control group nil treatment was imposed, and the model control group and BMSCs intervention group were established for further study on the EAU model. Upon completion of the EAU model, the BMSCs intervention group and model control group were subject to injection of 0.1 ml BMSCs suspension （2×10^6/ml） and 0.1 ml PBS into the vitreous cavity bilaterally, respectively. This entailed comparison on the ocular manifestations, histopathological grading and the level of serum/aqueous humor tu-mor necrosis factor-or （TNF-α）. Results The incubated cells appeared spindle-like and were immunologically pre-sented as CD105（＋）, CD34（-） and CD45（-） by flow cytometry. Normal control group did not evidence any aberrance in the anterior segment, the fundus, uhrasonographic presentations and HE staining. BMSCs intervention group yielded markedly attenuated inflammation and histopathological grading scores of the anterior segment than those in the model control group （all P〈0.05）. In the model control group the level of TNF-α in serum/aqueous humor was pronouncedly increased compared with that in normal control group （P〈0.05）. BMSCs intervention group was associated with a lower level of TNF-α in serum/aqueous humor compared with the model control group （P〈0.05）. Conclusion BMSCs have a therapeutic effect on rabbit EAU, possibly via the reduction in the levels of serum/aqueous humor inflammatory cy-tokines.