人脐带间充质干细胞对兔自身免疫性干眼的疗效研究

Treatment effect of mesenchymal stem cells on rabbit autoimmune dry eye

目的通过建立稳定的兔自身免疫性干眼模型,观察人脐带来源的间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)对自身免疫性干眼的作用并初步探讨其机制。方法体外分离、培养hUC-MSCs并对其进行鉴定;采用静脉回输自体激活的淋巴细胞建立兔自身免疫性干眼模型,将兔分为正常对照组、干眼模型组和h UCMSCs治疗组,h UC-MSCs治疗组于回输淋巴细胞后输注1 mL 1×10~7个·mL^(-1)hUCMSCs,连续输注5 d,对照组输注等体积的PBS。三组分别在术后2周、4周、6周检测泪液分泌量、泪膜破裂时间和角膜荧光素钠染色,6周后行组织病理学观察,并检测泪腺组织中细胞因子的表达水平。结果术后2周、4周、6周h UC-MSCs治疗组较干眼模型组泪液分泌量、泪膜破裂时间显著升高(均为P<0.05);角膜荧光素钠染色着色显著减轻,组织病理学观察泪腺组织淋巴细胞浸润较干眼模型组显著减少。hUC-MSCs治疗组和干眼模型组相比,泪腺组织中炎性因子IFN-γ、IL-17表达显著降低(F=50.48、18.13;均为P<0.05),而抑炎因子IL-10、TGF-β的表达显著升高(F=31.06、21.40;均为P<0.05)。结论h UC-MSCs减轻兔自身免疫性干眼,对其发挥免疫调控作用。

Objective To observe the effects of human umbilical cord derived mesenchymal stem cells（hUC-MSCs） on autoimmune dry eye, and explore the mechanism of the human umbilical cord derived mesenchymal stem cells. Methods The isolation and culture of human umbilical cord derived mesenchymal stem cells in vitro, and then identified;The autologous activated lymphocytes through vein was used to establish autoimmune dry eye rabbit model,and divided into normal control group, dry eye model group ,and mesenchymal stem cell treatment group. The mesenchymal stem ceils （ 1 × 10^7 cell · mL -1,1 mL） were infused in the treatment group after reinfusion of lymphocyte for 5 days, the control groups were infused with equal volume of PBS. In the three groups,tear secretion and tear film stability and corneal fluorescein staining were detected at 2 weeks,4 weeks and 6 weeks,histological observation was performed at 6 weeks, and the expression of cytokines in lacrimal gland tissue was detected. Results The tear secretion, tear break-up time in the treatment group were higher than those in the model group （ all P 〈 0.05 ）, corneal fluorescein staining was reduced, and the lymphocytes infiltration was obviously decreased. Compared with model group, the expression of IFN-gamma and IL-17 in the lacrimal gland tissue of treatment group were obviously decreased （F = 50.48,18.13 ; all P 〈 0.05 ）, but the expression of IL-10 and TGF-beta were increased （ F = 31.06,21.40 ; all P 〈 0.05 ）. Conclusion The mesenchymal stem ceils derived from human umbilical cord blood stem ceils can reduce their own immune dry eye,and play the role of immune regulation.