结核杆菌热休克蛋白肽段对胶原诱导性关节炎保护作用的研究

The protection of mycobacterial heat shock protein peptides against collagen-induced arthritis

目的:研究结核杆菌HSP65肽段(P256-270)和结核杆菌HSP70肽段(P111-125)对类风湿关节炎动物模型——胶原诱导性关节炎(CIA)大鼠的保护性作用。方法:结核杆菌HSP肽段滴鼻免疫大鼠,观察其对CIA的防治效果,观察指标包括关节炎指数评分、关节病理学分析、酶联免疫斑点实验(ELISPOT)检测脾脏淋巴细胞分泌细胞因子(IL-4、IFN-γ)水平、流式细胞术分析外周血T淋巴细胞亚群及酶联免疫吸附实验(ELISA)测定血清中抗CⅡ抗体水平。结果:分别经结核杆菌HSP65和HSP70肽段滴鼻免疫的实验组大鼠,与阳性对照组比较,关节炎指数明显下降(P<0·05),病理变化减轻,抑制性细胞因子IL-4水平升高(分别为P<0·05和P<0·01),而炎性细胞因子IFN-γ水平降低(P<0·01),CD4+/CD8+T细胞比值降低(P<0·05),血清中抗CⅡ抗体水平也降低(分别为P<0·05和P<0·01)。结论:应用结核杆菌HSP65肽段或HSP70肽段能减轻CIA大鼠的症状和体征,其作用机制与调节T细胞的功能有关,此研究可为探讨人类RA新的免疫治疗方法提供依据。

Objective： To research the protective effects of mycobacterial HSP65 peptide （P256-270） and mycobacterial HSP70 peptide（ P111-125 ） on the animal model of rheumatoid arthritis collagen induced arthritis （CIA）. Methods： Mycobacterial HSP peptides were admlnisterod intranasally to rats, then the protective effects on CIA rats were evaluated, including arthritis index, articular pathologic analysis ,cytokines（IL-4,IFN-T） production from lymphocytes of spleen by enzyme-linked immunespot（ELISPOT） ,T lymphocytes subsets analysis in peripheral blood by flow cytometry, levels of anti--C Ⅱ antibodies in serum detected by enzyme-linked immunoadsorhent assay（ELISA）. Results：The protective effects of nasal administration of mycobacterlal HSP65 peptide or HSP70 peptide to rats were observed. Compared with positive control group, arthritis index decreased （P 〈 0. 05）, pathologic changes alleviatod, levels of IL-4 increased（P〈0.05,P 〈0.01） and IFN-γ decresaed（P 〈0.01）, the rates of CD4^＋/CD8^＋T lymphecytes decreased（P〈 0. 05 ）, levels of anti---C Ⅱ antibodies in serum decreased （ P 〈 0. 05, P 〈 0. 01 ）. Conclusion： Nasal administration of mycobacterial lISP peptides can allevlate the symptoms and physical signs of CIA rats,and the mechanism is in connection with adjusting the function of T lymphocytes . The research provides new evidence for immunotherapy of human RA.