IL-6受体对类风湿关节炎成纤维样滑膜细胞致病信号及功能的抑制作用

Suppression of Malignant Signaling and Behavior of Fibroblast-like Synoviocytes from Rheumatoid Arthritis by Interleukin 6 Receptor Mimotope

关键词:  类风湿关节炎  成纤维样滑膜细胞  模拟表位肽  白介素-6受体

Key words:  rheumatoid arthritis  fibroblast-like synoviocyte  mimotope  IL-6R

摘要:

目的 观察白介素-6受体（IL-6R）模拟表位肽活化免疫后产生的抗体结合类风湿关节炎成纤维样滑膜细胞（RA-FLS）所表达的相应受体对相关致病信号及功能的抑制作用。方法 化学法制备模拟表位肽，后皮下免疫Wistar大鼠，取血清纯化抗体。酶联免疫吸附法评估血清抗体效价，免疫印迹法和免疫荧光法观察模拟表位肽诱导的抗体是否能结合IL-6R并促进其降解，免疫印迹法观察抗体对IL-6下游信号是否有抑制作用，应用CCK8试剂盒和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法观察诱导的抗体对细胞增殖和凋亡能力的影响。结果 IL-6R模拟表位肽免疫动物后获得的抗血清能够有效结合重组IL-6R，该血清经纯化后获得的抗体能够有效结合RA-FLS所表达的自然IL-6R并促进其降解，能进一步抑制下游信号通路、抑制RA-FLS的增殖，下调RA-FLS所具有的抗凋亡能力。结论 IL-6R受体模拟表位肽为潜在的控制RA-FLS恶性行为的多肽药物。

Abstract:

OBJECTIVE To observe whether the active immunization induced by interleukin-6 receptor (IL-6R) mimotope could bind to the IL-6R, inhibit the signalings and pathogenic behaviors of fibroblast-like synoviocytes from rheumatoid arthritis (RA-FLS). METHODS Wistar rats were immunized subcutaneously with IL-6R mimotope prepared chemically. The poly antibodies were purified from serum. Enzyme linked immunosorbent assay was performed to evaluate the titer of antisera. Western blot and immunofluorescence staining were performed to observe whether the antibodies purified could bind to IL-6R and induce their degradation. Western blot was performed to observe the inhibition of IL-6R. CCK-8 kit and TdT-mediated dUTP Nick-End Labeling were performed to evaluate the proliferation and apoptosis of RA-FLS. RESULTS Antibodies induced by IL-6R mimotope could bind to the IL-6R, induce their degradation, inhibit the signaling pathways and suppress the proliferation and anti-apoptosis of RA-FLS. CONCLUSION The IL-6R mimotope can be a potential agent for suppression of RA-FLS.