间充质干细胞联合环孢霉素A对大鼠角膜移植排斥的效果分析及炎症反应因子的参与作用研究
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摘要
目的探讨间充质干细胞联合环孢霉素A对角膜移植排斥的效果及炎症反应因子的参与作用。方法采用远交系Wistar雌性大鼠和SD雌性大鼠建立穿透性角膜移植排斥模型。随机分为空白对照组(尾静脉及肌肉分别注射不含药物的PBS,0.1 ml/kg和1 ml/kg),角膜移植排斥组(尾静脉及肌肉分别注射不含药物的PBS,0.1 ml/kg和1ml/kg)、治疗高剂量组[尾静脉注射MSCs(5×106)+肌注Cs A 10 mg/(kg·d)]、中剂量组[尾静脉注射MSCs(5×106)+肌注Cs A 5 mg/(kg·d)]、低剂量组[尾静脉注射MSCs(5×106)+肌注Cs A 2 mg/(kg·d)],每组8只。观察各组大鼠治疗后的角膜植片存活情况,并检测血清中相关的炎症反应蛋白水平。结果与空白对照组比较,角膜移植排斥组大鼠的角膜植片存活时间明显缩短(P<0.01),而治疗组大鼠的存活时间明显延长,且存在剂量依赖性(P<0.05,P<0.01);角膜移植排斥组大鼠炎症反应因子白介素细胞(IL)-2、IL-6、IL-10、纤维介素蛋白2和单核细胞趋化蛋白1、转化生长因子(TGF-β)、Smad1及Smad3、内质网跨膜蛋白肌醇酶1(IRE1)及其下游转录因子(XBP1)水平明显高于对照组(P<0.01),而治疗组大鼠的上述炎症因子水平明显降低,且存在剂量依赖性(P<0.05,P<0.01)。结论间充质干细胞联合环孢霉素A对角膜移植排斥有良好的治疗效果,且其作用可能与调控体内炎症反应因子水平异常有关。
Objective To explore the effect of mesenchymal stem cell combined Cs A on corneal transplantation and its regulations on inflammatory factors. Methods The penetrating corneal graft rejection model were prepared with female Wistar and SD rats. A penetrating keratoplasty rejection model was established using outbred Wistar female rats and female SD rats. The blank control group,corneal graft rejection group( established corneal graft rejection model),high dose group,middle dose group,low dose group[corneal transplant rejection model + postoperative injection of mesenchymal stem cells + intramuscular injection of cyclosporin A 10/5/2 mg/( kg·d) ] with 8 in each group were established. The penetrating corneal graft rejection model received mesenchymal stem cell combined Cs A treatment at different concentration. The concentration of inflammatory factors was detected at each group rats. Results The survive time of penetrating corneal graft rejection rat were prolonged significantly after treatment. The concentration of inflammatory factors including IL-2,IL-6,IL-10,TGF-β,Smad1,Smad3,IRE1 and XBP1 were increased greatly in model group rats when compared with control. And at the same time,those abnormalities were normalized dramatically after the treatment of mesenchymal stem cell combined Cs A with statistical differences. Conclusion Mesenchymal stem cell combined Cs A are effective on penetrating corneal graft rejection rats which maybe related with the regulation of inflammatory reactions.
Objective To explore the effect of mesenchymal stem cell combined Cs A on corneal transplantation and its regulations on inflammatory factors. Methods The penetrating corneal graft rejection model were prepared with female Wistar and SD rats. A penetrating keratoplasty rejection model was established using outbred Wistar female rats and female SD rats. The blank control group,corneal graft rejection group( established corneal graft rejection model),high dose group,middle dose group,low dose group[corneal transplant rejection model + postoperative injection of mesenchymal stem cells + intramuscular injection of cyclosporin A 10/5/2 mg/( kg·d) ] with 8 in each group were established. The penetrating corneal graft rejection model received mesenchymal stem cell combined Cs A treatment at different concentration. The concentration of inflammatory factors was detected at each group rats. Results The survive time of penetrating corneal graft rejection rat were prolonged significantly after treatment. The concentration of inflammatory factors including IL-2,IL-6,IL-10,TGF-β,Smad1,Smad3,IRE1 and XBP1 were increased greatly in model group rats when compared with control. And at the same time,those abnormalities were normalized dramatically after the treatment of mesenchymal stem cell combined Cs A with statistical differences. Conclusion Mesenchymal stem cell combined Cs A are effective on penetrating corneal graft rejection rats which maybe related with the regulation of inflammatory reactions.
引文
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[15]王莹莹.Smad3敲除有效改善小鼠心脏移植排斥反应[D].杭州:浙江大学,2016.
[16]王炯,周炼红,邢怡桥,等.TGF-β1对大鼠高危角膜移植排斥反应及超微结构的影响[J].眼科新进展,2010,30(4):313-317.
[17]李伟光,张博,吴波.转化生长因子β在器官移植排斥反应中的研究进展[J].医学综述,2014,20(9):1559-1561.
[18]白霄,宋光民,王维,等.单核细胞趋化蛋白-1和NF-κB通路在心脏移植排斥反应中的作用[J].山东大学学报(医学版),2013,51(11):1-5.
[19]张振瑜,吴京,马明,等.纤维介素蛋白2(fgl-2)在大鼠角膜移植排斥反应中的作用[J].眼科新进展,2015,35(10):913-917.
[20]王润芝,吴皓,刘一鸣,等.阻断kupffer细胞IRE1-XBP1通路对大鼠肝移植排斥反应的影响[J].第三军医大学学报,2016,38(22):2431-2437.
[2]Frigo AC,Fasolo A,Capuzzo C,et al.Corneal transplantation activity over 7 years:Changing trends for indications,patient demographics and surgical techniques from the corneal transplant epidemiological study(cortes)[J].Transplant Proc,2015,47(2):528-535.
[3]张迪,张红.角膜移植排斥基因治疗的研究进展[J].医学综述,2016,8(2):216-220.
[4]钟敬祥,张伟,赵伟,等.环孢霉素A微粒体抑制角膜移植排斥反应的临床观察[J].暨南大学学报(自然科学与医学版),2005,26(2):224-226,231.
[5]朱映芳,周志凌.FK506和环孢霉素A在兔角膜缘移植术后抑制免疫排斥反应的疗效比较[J].湘南学院学报(医学版),2008,10(4):4-6.
[6]Onishi R,Ohnishi S,Higashi R,et al.Human amnion-derived mesenchymal stem cell transplantation ameliorates dextran sulfate sodium-induced severe colitis in rats[J].Cell Transplant,2015,24(12):2601-2614.
[7]Zhang M,Liu D,Li S,et al.Bone marrow mesenchymal stem cell transplantation retards the natural senescence of rat hearts[J].Stem Cells Transl Med,2015,4(5):494-502.
[8]Kourtzelis I,Kotlabova K,Lim JH,et al.Developmental endothelial locus-1 modulates platelet-monocyte interactions and instant bloodmediated inflammatory reaction in islet transplantation[J].Thromb Haemost,2016,115(4):781-788.
[9]Edword JH,Chan Chi-Chao,Richard PW,et al.Clinical and immunological studies of comeal rejection in the rat penetrating keratoplasty model[J].Comea,1991,10(8):374-376.
[10]张杰,张环,唐滔,等.白细胞介素10和γ干扰素基因表达在大鼠肝移植排斥反应诊断中的意义[J].中国医学创新,2017,14(2):17-20.
[11]吕向微,李芳,王立明.白细胞介素10与肝移植急性排斥反应研究进展[J].医学综述,2013,19(3):433-436.
[12]孙曙,李州利,蔡明,等.白细胞介素17在肾移植排斥反应中的表达及意义[J].中国组织工程研究,2014,18(51):8275-8280.
[13]王平,涂同涛,臧思思,等.心脏移植排斥反应患者血清IL-6检测的意义[J].中国实验诊断学,2015,19(10):1673-1675.
[14]张振瑜.纤维介素蛋白2(FGL-2)及IL-23/IL-17在大鼠角膜移植排斥反应中的作用[D].广州:南方医科大学,2016.
[15]王莹莹.Smad3敲除有效改善小鼠心脏移植排斥反应[D].杭州:浙江大学,2016.
[16]王炯,周炼红,邢怡桥,等.TGF-β1对大鼠高危角膜移植排斥反应及超微结构的影响[J].眼科新进展,2010,30(4):313-317.
[17]李伟光,张博,吴波.转化生长因子β在器官移植排斥反应中的研究进展[J].医学综述,2014,20(9):1559-1561.
[18]白霄,宋光民,王维,等.单核细胞趋化蛋白-1和NF-κB通路在心脏移植排斥反应中的作用[J].山东大学学报(医学版),2013,51(11):1-5.
[19]张振瑜,吴京,马明,等.纤维介素蛋白2(fgl-2)在大鼠角膜移植排斥反应中的作用[J].眼科新进展,2015,35(10):913-917.
[20]王润芝,吴皓,刘一鸣,等.阻断kupffer细胞IRE1-XBP1通路对大鼠肝移植排斥反应的影响[J].第三军医大学学报,2016,38(22):2431-2437.