阻断CD40-CD40L信号通路对小鼠原位气管移植术后早期气道上皮细胞的影响

Effects of blocking CD40-CD40L pathway on the airway epithelial cells in the early stage after orthotopic tracheal transplantation in mice

目的本研究利用小鼠原位气管移植模型,应用anti m CD40L阻断CD40-CD40L信号通路,探讨其对气管移植模型中IL-17A介导的免疫反应的机制及其调控对移植物的影响,进一步解析其对移植物损伤和疾病预后的影响,为临床寻求新的预防和治疗方法。方法建立小鼠原位气管移植的模型,选取野生型Balb/c小鼠作为供体,野生型C57BL/6小鼠作为供体或受体。移植术后第3天,利用实时荧光定量PCR技术检测各组移植气管中IL-17A m RNA表达水平、ELISA检测血清中IL-17的含量;移植术后第15天,采用苏木精-伊红(HE)、Masson染色,观察各组移植气管组织的病理学改变。结果注射anti m CD40L(MR-1)抗体后,术后3 d移植物中IL-17A m RNA相对于同种异型非特异性免疫球蛋白抗体组有所下降(P<0.05);各组血清中IL-17的含量变化不明显(P>0.05)。术后15 d病理结果显示注射anti m CD40L后可以有效改善气管上皮细胞的损伤。结论阻断CD40-CD40L信号通路能够缓解原位气管移植术后IL-17A对气管上皮细胞的损伤。

This study was designed to explore the effects of blocking CD40-CD40 L pathway by anti m CD40 Lm Ab on graft in mouse orthotopic tracheal transplantation model, and explore new clinical prevention and treatmentmethods. The orthotopic tracheal transplantation model was constructed, and wild-type Balb/c mice were used asdonors, while wild-type C57BL/6 mice were used as donor or recipients. The expression level of IL-17 A m RNA intracheal was quantitated by real time quantitative PCR, and the protein level of IL-17 A in serum was measured byELISA 3 days after transplantation. Pathological changes of tracheal were examined by HE staining and Massonstaining 15 days after transplantation. Administration of anti m CD40 L m Ab significantly reduced IL-17 Atranscription in tracheal compared to non-specific immunoglobulin antibody treated group, while there was littleeffect on levels of IL-17 expression in serum. Pathology results showed that administration of anti m CD40 L m Abimproved tracheal epithelial cell injury. Taken together, blocking CD40-CD40 L pathway by anti m CD40 L m Ab canalleviate the damage of tracheal epithelial cells mediated by IL-17 A after orthotopic tracheal transplantation.