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

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,0.1ml/kg和1 ml/kg),角膜移植排斥组(尾静脉及肌肉分别注射不合药物的PBS,0.1 ml/kg和1ml/kg)、治疗高剂量组[尾静脉注射MSCs(5×106)+肌注CsA 10 mg/(kg·d)]、中剂量组[尾静脉注射MSCs(5×106)+肌注CsA5 mg/(kg·d)]、低剂量组[尾静脉注射MSCs(5×106)+肌注CsA 2mg/(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 CsA 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 CsA 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 CsA with statistical differences.Conclusion Mesenchymal stem cell combined CsA are effective on penetrating corneal graft rejection rats which maybe related with the regulation of inflammatory reactions.