CAJ | 학술논문

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。
목적：탐토오소사Ⅱ형효원단백（Zaocys typeⅡcollagen, ZCⅡ）대효원유도성관절염（Collagen-induced arthritis, CIA）소서장계막림파결림파세포（mesenteric lymph node lymphocytes, MLNLs）Treg/Th17세포급기세포인자적영향。방법계Ⅱ형효원단백소서미근부피하주사，유도CIA모형；위단백매소화법제취ZCⅡ，구복고、중、저제량ZCⅡ간예。시각평분법평개관절종창，HE염색평개슬관절조직병이학；분리소서MLNLs，류식세포술（FCM）검측Treg화Th17세포적비솔；매련면역흡부측정（ELISA）법검측MLNLs배양상청중TGF-β화IL-17적활성。결과여정상조비교，CIA소서관절염급조직병이학평분승고（P<0.05），MLNLs중Treg화Th17적비솔급기분비TGF-β화IL-17수평상승（P<0.05）。여CIA모형조비교，불동제량ZCⅡ간예조관절염급조직병이학평분강저（P<0.05），MLNLs중Treg세포적비솔상승，Th17세포적비솔하강（P<0.05），MLNLs분비TGF-β수평상승，분비IL-17수평하강（P<0.05）。결론구복ZCⅡ능구개선CIA소서관절염급조직병이학평분；ZCⅡ가능통과조절MLNLs중Treg/Th17비솔급기세포인자적수평，중신유도CIA소서적면역평형，완해관절염증。
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<0.05). In ZCII peptide-treated mice, the scores for arthritis and histopathological changes were significantly lower than those in CIA model group (P<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<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.