乌梢蛇Ⅱ型胶原蛋白调控胶原诱导性关节炎小鼠肠系膜淋巴结Treg/Th17平衡  

Zaocys type Ⅱ collagen regulates mesenteric lymph node Treg/Th17 cell balance in mice with collagen-induced arthritis

作　　者：王浩[1] 冯知涛[1] 朱俊卿[1] 李娟[1] 

机构地区：[1]南方医科大学南方医院中医科,广东广州510515

出　　处：《南方医科大学学报》2014年第5期622-626,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81173456);国家教育部博士点基金(20094433110006);广东省自然科学基(S2012010009164);广东省"211"工程三期重点学科建设项目~~

摘　　要：目的探讨乌梢蛇Ⅱ型胶原蛋白(Zaocys typeⅡcollagen,ZCⅡ)对胶原诱导性关节炎(Collagen-induced arthritis,CIA)小鼠肠系膜淋巴结淋巴细胞(mesenteric lymph node lymphocytes,MLNLs)Treg/Th17细胞及其细胞因子的影响。方法鸡Ⅱ型胶原蛋白小鼠尾根部皮下注射,诱导CIA模型;胃蛋白酶消化法提取ZCⅡ,口服高、中、低剂量ZCⅡ干预。视觉评分法评价关节肿胀,HE染色评价膝关节组织病理学;分离小鼠MLNLs,流式细胞术(FCM)检测Treg和Th17细胞的比率;酶联免疫吸附测定(ELISA)法检测MLNLs培养上清中TGF-β和IL-17的活性。结果与正常组比较,CIA小鼠关节炎及组织病理学评分升高(P<0.05),MLNLs中Treg和Th17的比率及其分泌TGF-β和IL-17水平上升(P<0.05)。与CIA模型组比较,不同剂量ZCⅡ干预组关节炎及组织病理学评分降低(P<0.05),MLNLs中Treg细胞的比率上升,Th17细胞的比率下降(P<0.05),MLNLs分泌TGF-β水平上升,分泌IL-17水平下降(P<0.05)。结论口服ZCⅡ能够改善CIA小鼠关节炎及组织病理学评分;ZCⅡ可能通过调节MLNLs中Treg/Th17比率及其细胞因子的水平,重新诱导CIA小鼠的免疫平衡,缓解关节炎症。Objective To investigate the effect of oral administration of Zaocys type II collagen （ZCII） on the percentages of Treg/Th17 cells in mesenteric lymph node lymphocytes （MLNLs） in mice with collagen-induced arthritis （CIA）. Methods CIA was induced in male C57BL/6 mice by immunization with chicken type II collagen. Three weeks later, ZCII, purified by pepsin digestion, was orally administered in the mice for 7 consecutive days （daily dose of 10, 20, or 40 μg/kg）. The severity of arthritis in each limb was evaluated using a macroscopic scoring system, and histopathological changes of the joint were observed microscopically with HE staining. The percentages of Treg and Th17 cells in MLNLs was detected by flow cytometry, and the levels of transforming growth factor-β（TGF-β） and interleukin-17 （IL-17） in the supernatant of MLNLs were measured by enzyme-linked immunosorbent assay. Results Compared with normal control mice, the mice with CIA had significantly higher scores for arthritis and histopathological changes, with also significantly increased percentages of Treg and Th17 cells in MLNLs and elevated levels of TGF-β and IL-17 in MLNL supernatant （P＆lt;0.05）. In ZCII peptide-treated mice, the scores for arthritis and histopathological changes were significantly lower than those in CIA model group （P＆lt;0.05）, and Treg cell percentage in MLNLs was up-regulated while Th17 cell percentage lowered;the level of TGF-βwas increased but IL-17 was decreased significantly （P＆lt;0.05）. Conclusion Oral administration of ZCII improves CIA in mice by regulating the percentages of Treg/Th17 cells and the cytokine levels in MLNLs, suggesting the value of ZCII as a promising candidate agent for treatment of rheumatoid arthritis.

关 键 词：Ⅱ型胶原蛋白 类风湿关节炎 胶原诱导性关节炎 TREG Th17 口服免疫耐受 

分 类 号：R593.22[医药卫生—内科学]