【摘要】 目的:观察骨髓间充质干细胞（bone marrow mesenchymal stem cells,BMSCs）移植对胶原诱导性关节炎（collagen induced arthritis,CIA）大鼠血清细胞因子的影响。方法:选取清洁级雌性Wistar大鼠60只。用颈椎脱臼断颈法处死1只大鼠,取股骨和胫骨骨髓,分离、培养BMSCs。其余59只大鼠采用随机数字表分为正常对照组（20只）、模型组（20只）、细胞移植组（19只）。模型组和细胞移植组每只大鼠在尾根部、背部及左后足跖部皮下注射含牛Ⅱ型胶原的乳化液共0.5 mL,正常对照组注射等量生理盐水。于第1次注射后第14天再注射1次。于第2次注射后的第15天,各组分别随机处死2只大鼠,观察左侧踝关节组织形态,判断造模是否成功。于第2次注射的当天,细胞移植组大鼠通过尾静脉注射BMSCs悬液,正常对照组和模型组注射等量生理盐水。干预30 d后,采用关节炎指数（arthritis index,AI）评价各组大鼠关节炎严重程度;采用ELISA法检测大鼠血清中血管内皮生长因子（vascular endothelial growth factor,VEGF）、基质金属蛋白酶3（matrix metalloproteinase 3,MMP-3）、肿瘤坏死因子α（tumor necrosis factor,TNF-α）、白细胞介素17（interleukin-17,IL-17）水平。结果:①模型鉴定结果。造模完成后的第15天,正常对照组大鼠踝关节结构完整,组织切片未见明显炎性细胞增殖;模型组和细胞移植组大鼠踝关节滑膜增生,组织切片可见大量增殖的炎性细胞,提示造模成功。②关节炎严重程度评价结果。干预30 d后,正常对照组、模型组和细胞移植组AI值分别为（1.15±0.32）分、（8.25±3.18）分和（5.07±1.29）分,3组间的总体差异有统计学意义（F=2.823,P=0.005）。模型组和细胞移植组AI值均高于正常对照组（P=0.017,P=0.023）,细胞移植组AI值低于模型组（P=0.042）。③血清细胞因子水平检测结果。干预30 d后,3组间血清VEGF、MMP-3、TNF-α和IL-17水平总体比较,差异均有统计学意义[（10.45±3.44）pg·mL^-1,（22.43±6.49）pg·mL^-1,（17.22±5.20）pg·mL^-1,F=2.059,P=0.043;（5.39±1.27）pg·mL^-1,（12.49±3.22）pg·mL^-1,（7.30±2.11）pg·mL^-1,F=2.823,P=0.005;（22.34±8.50）pg·mL^-1,（44.39±12.29）pg·mL^-1,（35.55±9.40）pg·mL^-1,F=1.983,P=0.048;（5.23±1.28）pg·mL^-1,（11.26±3.07）pg·mL^-1,（8.34±4.30）pg·mL^-1,F=2.557,P=0.015]。模型组和细胞移植组血清VEGF、MMP-3、TNF-α和IL-17水平均高于正常对照组（模型组和正常对照组比较:P=0.022,P=0.040,P=0.033,P=0.016;细胞移植组和正常对照组比较:P=0.019,P=0.031,P=0.022,P=0.047）;细胞移植组血清VEGF、MMP-3、TNF-α和IL-17水平低于模型组（P=0.044,P=0.012,P=0.033,P=0.048）。结论:BMSCs移植可减轻CIA大鼠关节炎严重程度,降低CIA大鼠血清中VEGF、MMP-3、TNF-α和IL-17水平。更多还原

【Abstract】 Objective:To observe the effects of bone marrow mesenchymal stem cells(BMSCs)transplantation on serum cytokines in rats with collagen-induced arthritis(CIA).Methods:Sixty clean-grade female Wistar rats were selected.One rat was randomly selected out from the 60 rats and was executed by using cervical dislocation method,and its bone marrow were fetched out from femur and tibia for separating and culturing BMSCs.The other 59 rats were randomly divided into normal control group(20),model group(20)and BMSCs transplantation group(19)by using random digits table.The rats in model group and BMSCs transplantation group were subcutaneously injected with emulsified liquid containing bovine typeⅡcollagen with dose of 0.5 mL into their tail roots,backs and left posterior plantar areas,while the rats in normal control group were subcutaneously injected with the same dose of normal saline(NS)in the same places.The second subcutaneous injection was performed on rats at the 14 th day after the first injection.At 15 th day after the second injection,2 rats were randomly selected out from each group and were executed,and their left ankle-joints were fetched out and the tissue morphology of left ankle-joints was observed to determine whether the models were built successfully.On the day of the second injection,the rats in BMSCs transplantation group were injected with BMSCs suspension into caudal vein,while the rats in normal control group and model group were injected with the same dose of NS into caudal vein.After 30-day intervention,the severity of arthritis was evaluated by using arthritis index(AI),and the serum levels of vascular endothelial growth factor(VEGF),matrix metalloproteinase 3(MMP-3),tumor necrosis factor(TNF-α)and interleukin-17(IL-17)were measured by using ELISA method.Results:At the 15 th day after the modeling,the structure of ankle-joint was intact and no obvious proliferative inflammatory cells were found in tissue sections in rats of normal control group,while synovial hyperplasia was found in ankle joint and a large number of proliferative inflammatory cells were found in tissue sections in rats of model group and BMSCs transplantation group,which suggested that the models were successfully built.After 30-day intervention,the AI values of normal control group,model group and BMSCs transplantation group were 1.15+/-0.32,8.25+/-3.18 and 5.07+/-1.29 points respectively,and there was statistical difference in AI values between the 3 groups in general(F=2.823,P=0.005).The AI values were higher in model group and BMSCs transplantation group compared to normal control group,and were lower in BMSCs transplantation group compared to model group(P=0.017,P=0.023;P=0.042).After 30-day intervention,there was statistical difference in serum levels of VEGF,MMP-3,TNF-α and IL-17 between the 3 groups in general(10.45+/-3.44,22.43+/-6.49,17.22+/-5.20 pg/mL,F=2.059,P=0.043;5.39+/-1.27,12.49+/-3.22,7.30+/-2.11 pg/mL,F=2.823,P=0.005;22.34+/-8.50,44.39+/-12.29,35.55+/-9.40 pg/mL,F=1.983,P=0.048;5.23+/-1.28,11.26+/-3.07,8.34+/-4.30 pg/mL,F=2.557,P=0.015).The serum levels of VEGF,MMP-3,TNF-α and IL-17 were higher in model group and BMSCs transplantation group compared to normal control group,and were lower in BMSCs transplantation group compared to model group(P=0.022,P=0.040,P=0.033,P=0.016;P=0.019,P=0.031,P=0.022,P=0.047;P=0.044,P=0.012,P=0.033,P=0.048).Conclusion:BMSCs transplantation can reduce the severity of arthritis and decrease the serum levels of VEGF,MMP-3,TNF-α and IL-17 in rats with CIA.更多还原