大鼠喉移植术后移植喉组织TNF-α和IL-10表达的变化

Changes of TNF-αand IL-10 expression in transplanted laryngeal tissues during acute rejection after laryngeal transplantation in rats

目的:观察大鼠喉移植术后移植喉组织不同部位TNF-α和IL-10表达的变化,探讨其与急性排斥反应的关系。方法:进行Wistar→SD大鼠喉移植,术后依照注射环孢霉素A(CsA)剂量不同随机分为3组:0 mg CsA组、5 mg CsA组及10 mg CsA组,以未进行喉移植手术的SD大鼠为对照组。术后第3、7、11天取各组移植喉组织进行病理学观察,免疫组化检测TNF-α、IL～10在不同部位的表达变化。结果:0、5 mg CsA组在术后第3、7、11天分别符合轻、中、重度排斥,10 mg CsA组无明显排斥征象;免疫组化检测显示0、5、10 mg CsA组TNF-α、IL-10均阳性表达,对照组基本不表达。0、5、10mg CsA组喉组织黏膜上皮及黏膜下层TNF-α、IL-10阳性区面积比值均明显高于肌肉及软骨组织(P<0.01);0、5 mg CsA组TNF-α表达始终显著高于10mg CsA组(P<0.01).0、5 mg CsA组IL-10表达在第11天低于10mg CsA组(P<0.01)。结论:供体喉的高抗原性主要集中于喉的黏膜上皮及黏膜下层组织;喉移植术后TNF-α和IL-10在移植组织的表达变化与急性排斥反直的进程相关,检测其表达变化可用于预测喉移植急性排斥反应。

Objective：To explore the expression of TNF-α and IL-10 in different layers of laryngeal tissues after laryngeal transplantation and its relationship with acute rejection. Methods： Laryngeal heterotopic transplantation was performed in Wistar and SD rats （Wistar→SD rats）. The SD rats were divided into 4 groups： Group I ： Sham control （receive no transplantation）; Group Ⅱ （receive transplantation, without cyelosporine A treatment）; Group Ⅲ（receive transplantation, with 5 mg· kg^-1· d^-1 cyelosporine A treatment）; and Group Ⅳ （receive transplantation, with 10 mg · kg^-1 · d^-1 cyelosporine A treatment）. The transplanted larynx was harvested on 3, 7 and 11 days after transplantation for pathological examination. The expression of TNF-α and IL-10 in different layers of grafts was detected immunohistochemically. Results： Pathological observation showed mild, moderate and severe acute rejection in GroupⅡ and Ⅲ on 3, 7 and 11 days after transplantation, respectively; there was no obvious rejection in Group Ⅳ. Immunohistochemical staining showed expression of TNF-a and IL-10 in Group Ⅱ, Ⅲ , and Ⅳ, not in Group Ⅰ. The ratios of the positive areas of TNF-α and IL-10 in the mucosal and submucosal layers were significantly higher than those in the muscle and cartilage layers of laryngeal tissues （P〈0.01） in Group Ⅱ , Ⅲ , and Ⅳ. The expression levels of TNF-α in Group Ⅱ, Ⅲ were always significantly higher than that in Group Ⅳ （P〈0.01） ;the expression levels of IL-10 in Group Ⅱ , Ⅲ were lower than that in Group Ⅳ on 11 days after transplantation （P〈0.01）. Conclusion： The high-antigenicity of laryngeal graft mainly locates in the mucosal and submucosal layers. Post-laryngeal transplantation expression of TNF-α and IL-10 is correlated with the course of acute rejection; detection of the expression may be used in predicting early acute rejection after laryngeal transplantation.