CAJ | 학술논문

目的观察Ⅱ型胶原短肽(CⅡ260-272)对胶原诱导性关节炎(CIA)大鼠T淋巴细胞激活的影响,探讨其在类风湿关节炎(RA)发病中的作用.方法建立CIA大鼠模型,分别在建模1、2…8周后,体外分离、培养大鼠脾淋巴细胞,采用活细胞计数(CCK-8)法检测CⅡ260-272短肽对淋巴细胞增殖反应的影响,并分析CⅡ短肽特异性T淋巴细胞增殖反应与滑膜炎及血管翳评分、成模时间及抗CⅡ及其短肽抗体之间的相关性.用SPSS 17.0软件进行数据分析.滑膜炎和血管翳评分结果比较用单因素方差分析,方差齐用LSD法,方差不齐用Games-Howell法；2组间增殖水平比较用Mann-Whitney U检验；细胞增殖与其他指标的关系用Spearman相关分析；2组间抗体水平比较用t检验.结果免疫3周后,CⅡ短肽特异性T细胞即出现明显的增殖反应[0.963(0.332～1.628)],与0周组大鼠[0.332(0.331～0.357)]比较,差异有统计学意义(P＜0.05)；分析发现,T细胞增殖反应与成模时间及抗bCⅡ抗体、抗Cit-bCⅡ抗体水平均呈正相关(r分别为:0.624,0.413,0.575,P均＜0.01).免疫2周后模型组大鼠中抗CⅡ260-270抗体(0.827±0.155)与0周(0.043±0.009)比较,差异有统计学意义(P＜0.01),免疫4周后抗bCⅡ抗体(0.362±0.062)、抗Cit- bCⅡ抗体(0.390±0.141)与0周相比,均有显著升高(P＜0.01)；同时,这3种抗体水平与血管翳评分也有显著相关性(P＜0.01或P＜0.05).结论 CⅡ短肽在建模早期即可刺激CIA大鼠脾淋巴细胞异常增殖,并在血清中检出针对CⅡ短肽的抗体反应.提示在利用CⅡ分子诱发CIA疾病发生过程中,该表位肽在驱动T、B淋巴细胞活化及引起自身免疫反应中发挥了重要作用.
목적 관찰Ⅱ형효원단태(CⅡ260-272)대효원유도성관절염(CIA)대서T림파세포격활적영향,탐토기재류풍습관절염(RA)발병중적작용.방법 건립CIA대서모형,분별재건모1、2…8주후,체외분리、배양대서비림파세포,채용활세포계수(CCK-8)법검측CⅡ260-272단태대림파세포증식반응적영향,병분석CⅡ단태특이성T림파세포증식반응여활막염급혈관예평분、성모시간급항CⅡ급기단태항체지간적상관성.용SPSS 17.0연건진행수거분석.활막염화혈관예평분결과비교용단인소방차분석,방차제용LSD법,방차불제용Games-Howell법；2조간증식수평비교용Mann-Whitney U검험；세포증식여기타지표적관계용Spearman상관분석；2조간항체수평비교용t검험.결과 면역3주후,CⅡ단태특이성T세포즉출현명현적증식반응[0.963(0.332～1.628)],여0주조대서[0.332(0.331～0.357)]비교,차이유통계학의의(P＜0.05)；분석발현,T세포증식반응여성모시간급항bCⅡ항체、항Cit-bCⅡ항체수평균정정상관(r분별위:0.624,0.413,0.575,P균＜0.01).면역2주후모형조대서중항CⅡ260-270항체(0.827±0.155)여0주(0.043±0.009)비교,차이유통계학의의(P＜0.01),면역4주후항bCⅡ항체(0.362±0.062)、항Cit- bCⅡ항체(0.390±0.141)여0주상비,균유현저승고(P＜0.01)；동시,저3충항체수평여혈관예평분야유현저상관성(P＜0.01혹P＜0.05).결론 CⅡ단태재건모조기즉가자격CIA대서비림파세포이상증식,병재혈청중검출침대CⅡ단태적항체반응.제시재이용CⅡ분자유발CIA질병발생과정중,해표위태재구동T、B림파세포활화급인기자신면역반응중발휘료중요작용.
Objective To examine the proliferative effect of synthetic C Ⅱ260-272 peptide on T cells of collagen-induced arthritis (CIA) rat,and to explore the role of C Ⅱ 260-272 in RA.Methods The rat model of collagen-induced arthritis (CIA) was established.The T lymphocytes were isolated and incubated with C Ⅱ 260-272 peptide.Proliferation of T cells was determined by cell counting kit-8.The data were analyzed with SPSS 17.0.The Mann-Whitney U test was used for proliferation levels comparison between the two groups,and the relationship of cell proliferation with other indicators was analyzed with Spearman's correlation.Antibody levels between the two groups were compared using t test.Results Three weeks after the first immunization,T cell response to C Ⅱ 260-272 in the CIA groups [0.963 (0.332-1.628)] was more significant than in controls [0.332 (0.331-0.357)](P＜0.05),and the response was associated with disease course (r=0.624,P＜0.01 ).Strong correlation between T cell proliferation and the antibodies,including antibC Ⅱ antibody and anti-Cit-bC Ⅱ antibody was also observed(P＜0.01 ).Two weeks later,the levels of anti-C Ⅱ peptide antibody in each CIA group were significantly higher than those in the controls (P＜0.01).The antibodies strongly correlated with pannus formation (P＜0.01,P＜0.05).Conclusion The proliferative response of T cell to C Ⅱ 260-272 peptide can be found in the early stage of CIA rat,and the anti-C Ⅱ peptide antibody could be detected in the serum,which suggeststhat the C Ⅱ peptide may play an important role in activating T/B lymphocytes and causing immune reaction when CIA is induced by C Ⅱ.