P2X7R抑制剂BBG对小鼠异基因造血干细胞移植后aGVHD的抑制作用研究

Effect of BBG on Acute Graft-Versus-Host Disease in Mice after Allogeneic Hematopoietic Stem Cell Transplantation

目的:探讨P2X7R抑制剂酸性亮蓝G(brilliant blue G,BBG)对小鼠异基因造血干细胞移植(allo-HSCT)后急性移植物抗宿主病(aGVHD)发生的影响。方法:建立小鼠allo-HSCT后并发aGVHD模型,通过不同剂量P2X7R抑制剂BBG(50和75 mg/kg)给药处理,观察受鼠生存情况、体重变化,检测肝脏病理变化及肝功能改变,检测肝脏P2X7R、NLRP3、Caspase-1、IL-1β、IL-18 mRNA及相关蛋白表达的变化。结果:成功建立小鼠allo-HSCT后并发aGVHD模型。腹腔注射BBG可减轻受鼠弓背、竖毛、皮肤剥脱、体重下降等aGVHD的临床表现;减轻受鼠肝脏水肿、出血、坏死等病理改变,改善受鼠的肝功能;降低受鼠肝脏P2X7R、IL-1β蛋白的表达;降低受鼠肝脏P2X7R、NLRP3、Caspase-1、IL-1β、IL-18因子mRNA表达;且aGVHD+75 mg/kg BBG组受鼠各项指标变化最为显著。结论:BBG可减轻allo-HSCT后aGVHD引起的肝脏炎性损伤,减少炎性因子分泌,且75 mg/kg BBG组的保护作用优于50 mg/kg BBG组。

Objective： To investigate the effect of P2X7R antagonist brilliant blue G（BBG） on aGVDH of mice after allo-HSCT. Methods： aGVHD mouse model after HSCT was established and treated with the P2X7R antagonist BBG of different dosages（50 mg/kg and 75 mg/kg）. After treatment,the survival,body weight,pathological and liver function of aGVDH mice were abserved,and the expression levels of P2 X7,NLRP3,caspase-1,IL-1β,IL-18 mRNA and protein were evaluated by real-time PCR and Western blot. Results：The allo-HSCT aGVHD mouse model was successfully established,the intraperitoneal injection of BBG alleviated the aGVHD clinical manifestations including roachback,ruffled fur,skin peeling and weight loss of recipient mice,decreased P2X7R and IL-1β expression and reduced the mRNA levels of P2 X7 R,NLRP3,Caspase-1,IL-1β and IL-18. Furthermore,GVHD group receiving75 mg/kg BBG showed most significant difference of these indexes. Conclusion： BBG alleviates liver inflammatory damage induced by aGVHD after allo-HSCT,and decreases the expression of proinflammatory cytokines. Moreover,the protective effect of that of BBG 75 mg/kg group is better than that of BBG 50 mg/kg group.