CAJ | 학술논문

目的观察奇壬醇对牛Ⅱ型胶原特异性淋巴细胞体内和体外的免疫调节作用,探讨奇壬醇对抗原特异性淋巴细胞的免疫抑制作用机制.方法将Wistar大鼠随机分为对照组、模型组、奇壬醇组和泼尼松龙组.除对照组外,均复制胶原诱导关节炎大鼠模型,造模当天开始给药.造模30 d后取脾脏和引流淋巴结,制备单细胞悬液,经体外培养48 h后,ELISA法检测培养上清中干扰素-γ(IFN-γ)、白介素-10(IL-10)、IL-4、肿瘤坏死因子-α(TNF-t)含量；在Ⅱ型胶原诱导下,取正常大鼠淋巴结细胞、脾细胞与不同浓度的奇壬醇共同培养,检测胶原诱导的淋巴细胞增殖及凋亡.结果与模型组相比,奇壬醇组脾细胞培养上清中IFN-γ和TNF-α含量降低(P＜0.05,P＜0.01),IL-10和IL-4水平升高(P＜0.05,P＜0.01)；淋巴结细胞培养上清中IFN-γ和TNF-α含量降低(P＜0.05,P＜0.001),IL-10和IL-4水平升高(P＜0.05,P＜0.001);体外实验中,奇壬醇抑制Ⅱ型胶原特异性淋巴结细胞增殖,促进Ⅱ型胶原特异性淋巴结细胞和脾细胞凋亡,均呈剂量依赖关系.结论奇壬醇可通过抑制Ⅱ型胶原特异性淋巴细胞分泌促炎因子,促进胶原特异性淋巴细胞分泌抗炎因子,抑制胶原特异性淋巴细胞增殖和促进其凋亡,以多条途径发挥免疫抑制作用.
목적 관찰기임순대우Ⅱ형효원특이성림파세포체내화체외적면역조절작용,탐토기임순대항원특이성림파세포적면역억제작용궤제.방법 장Wistar대서수궤분위대조조、모형조、기임순조화발니송룡조.제대조조외,균복제효원유도관절염대서모형,조모당천개시급약.조모30 d후취비장화인류림파결,제비단세포현액,경체외배양48 h후,ELISA법검측배양상청중간우소-γ(IFN-γ)、백개소-10(IL-10)、IL-4、종류배사인자-α(TNF-t)함량；재Ⅱ형효원유도하,취정상대서림파결세포、비세포여불동농도적기임순공동배양,검측효원유도적림파세포증식급조망.결과여모형조상비,기임순조비세포배양상청중IFN-γ화TNF-α함량강저(P＜0.05,P＜0.01),IL-10화IL-4수평승고(P＜0.05,P＜0.01)；림파결세포배양상청중IFN-γ화TNF-α함량강저(P＜0.05,P＜0.001),IL-10화IL-4수평승고(P＜0.05,P＜0.001);체외실험중,기임순억제Ⅱ형효원특이성림파결세포증식,촉진Ⅱ형효원특이성림파결세포화비세포조망,균정제량의뢰관계.결론 기임순가통과억제Ⅱ형효원특이성림파세포분비촉염인자,촉진효원특이성림파세포분비항염인자,억제효원특이성림파세포증식화촉진기조망,이다조도경발휘면역억제작용.
Objective To investigate the effect of kirenol on bovine type II collagen (CII)-specific lymphocytes in vivo and in vitro,and explore the mechanism of kirenol-induced immunosuppression in antigen-specific lymphocytes.Methods Twenty-four Wistar rats were randomized into control group,collagen-induced arthritis (CIA) model group,kirenol group (2 mg/kg),and prednisolone group (2 mg/kg).After CⅡ injection,the rats in the latter two groups received intragastric administration of kirenol and prednisolone for 30 days,and the spleens and draining lymph nodes of the rats were harvested to prepare single cell suspensions for measurement of the cytokine levels using ELISA.In the in vitro experiment,the lymphocytes from the control rats,with or without 20 μg/ml CII treatment in the presence of 0-80 μg/ml kirenol,were evaluated for cell proliferation and apoptosis using [3H]-thymidine incorporation and flow cytometry,respectively.Results Compared with those in CIA group,IFN-γ and TNF-α production was significantly reduced in splenocyte culture supeRNatant of kirenol group (P＜0.05 and P＜0.01,respectively),and the level of IL-10 and IL-4 was up-regulated (P＜0.05 and P＜0.01,respectively); IFN-γ and TNF-αsecretion by the cultured lymph node cells (LNCs) significantly decreased (P＜0.05 and P＜0.001,respectively) and IL-10 and IL-4 production increased (P＜0.05,P＜0.001) in kirenol group.In the in vitro experiment,kirenol treatment caused obvious suppression of CⅡ-induced LNC proliferation and dose-dependently induced antigen-specific apoptosis of the splenocytes and LNCs.Conclusion Kirenol treatment reduces pro-inflammatory cytokine secretion,increases anti-inflammatory cytokine production,inhibits cell proliferation and induces apoptosis of CII-specific lymphocytes in vitro,suggesting the potential of kirenol as an immunosuppressant.