免疫调节剂CH2b对葡萄糖－6－磷酸异构酶混合肽段诱导的类风湿性关节炎小鼠的影响

摘要

目的：探究前期合成的含噻唑烷－4－酮的免疫调节剂CH2b 对葡萄糖－6－磷酸异构酶（ Glucose－6－phosphate isomerase，GPI）混合肽段诱导的类风湿性关节炎（Rheumatoid arthritis，RA）小鼠发病的影响。方法：hGPI325－339、hGPI469－483多肽片段与完全弗氏佐剂充分乳化后，于DBA／1小鼠尾根部多点皮下注射，并用百日咳毒素加强免疫。然后分别用α－半乳糖神经酰胺（α－GalCer）和CH2b对模型小鼠进行干预，监测各组小鼠体重变化和足踝关节肿胀变化情况，关节组织苏木精－伊红（Hematoxylin－eosin staining，HE）染色法观察炎细胞浸润情况，流式细胞技术（Fluorescence－activated cell sorting，FACS）检测全血中iNKT细胞频率变化，微量样本多指标流式蛋白定量技术（Cytometric bead array，CBA）检测血清中细胞因子TNF－α、IL－6、IL－4、IFN－γ水平。结果：与GPI混合肽段诱导的模型组相比，α－GalCer和免疫调节剂CH2b均可显著改善模型小鼠的体重增长缓慢、关节肿胀情况（P＜0．05）；炎性细胞浸润减少；且二者均可以显著提高RA小鼠体内iNKT细胞的频率，与模型组相比，差异有统计学意义（P＜0．05）；α－GalCer、CH2b组可以降低模型小鼠炎症高峰期TNF－α、IL－6、IFN－γ水平，两组结果差异无统计学意义（P＞0．05）。结论：免疫调节剂CH2b通过激活iNKT细胞影响炎性细胞因子的分泌，缓解GPI混合肽段诱导的关节炎。%Objective:To explore the effect of the synthetic immunomodulator CH 2b with a thiazolidin-4-one ring on the pathogenesis of rheumatoid arthritis (RA) mice induced by glucose-6-phosphate isomerase(GPI) mixed peptides.Methods:hGPI325-339 ,hGPI469-483 peptide fragments were mixed with complete freund′s adjuvant fully ,DBA/1 mice were givien subcutaneous injection of the emulsifiers,pertussis toxin to strengthen immunity.And then RA mice were intervened with α-GalCer and CH2b,the changes of body weight ,ankle joint symptoms scores were observed .The joint tissues stained with hematoxylin and eosin ( HE) was used to evaluate the inflammatory cells.Fluorescence-activated cell sorting ( FACS) was used to detect the frequency changes of iNKT cells .Cytometric bead array(CBA) was used to analyze the levels of serum cytokines TNF-α,IL-6,IL-4,IFN-γ.Results: Compared with the model group,α-GalCer,CH2b could reduce the inflammation of the model mice ,significantly improve the body weight growth and the joint swelling(P<0.05).The inflammatory cells infiltration were decreased.More importantly,CH2b improved the frequency of iNKT cells in peak,the difference was statistically significant (P<0.05).The levels of TNF-α,IL-6,IFN-γin serum were significantly decreased(P<0.05) by CH2b.α-GalCer,CH2b could decrease the levels of TNF-α,IL-6,IFN-γ,and the two groups had no statistically significant difference(P>0.05).Conclusion:Immunomodulator CH2b by activating iNKT cells affect the secretion of inflammatory cytokines ,and it relieved the GPI induced arthritis .