【作者】 黄国祥；

【导师】 周文斌；

【作者基本信息】 中南大学 ， 神经病学， 2007， 硕士

【摘要】 目的：运用逆转录聚合酶链反应（RT-PCR）测定IL-23p19、IL-17、IL-6在慢性EAE模型C57BL/6小鼠脑组织中的动态表达变化，并用TGF-β1进行干预，观察其变化，以探讨IL-23/Th17细胞炎症轴在慢性EAE病程中的作用。方法：72只近交系清洁级8-10周健康雌性C57BL/6J小鼠，随机分为佐剂组、致病组、干预组和假干预组。各组小鼠又分为发病初、发病高峰和慢性期组。致病组用人工合成的MOG_35-55和自制的完全弗氏佐剂免疫C57BL/6J小鼠制成慢性EAE动物模型。干预组在免疫同时，每只小鼠注射TGF-β1。假干预组在免疫同时注入PBS作为干预组对照。佐剂组注入完全弗氏佐剂。按Benson评分标准做EAE严重性的临床评估。用RT-PCR技术检测佐剂对照组、致病组、干预组和假干预组发病初、发病高峰和慢性期组脑组织IL-17、IL-23p19、IL-6mRNA的表达变化。结果：1动物模型：慢性EAE致病组在第一次免疫后14-20天发病，20-24天达高峰，28-32天缓解，发病小鼠体重明显减轻；干预组体重减轻，13-17天发病，17-22天到达高峰，更易趋于缓解。假干预组临床表现和发病特点同致病组。佐剂组体重呈增加趋势，Benson标准评分为0分。2 RT-PCRIL-23p19、IL-6和IL-17mRNA在佐剂组呈低表达，在假干预组各期表达与致病组无差异（P＞0.05）。2．1致病组IL-23p19发病初期表达升高；高峰期表达有下降（两者相比有统计学意义，P＜0.05）；慢性期表达下降，但和高峰期无明显统计学差异（P＞0.05）。干预组IL-23p19表达，在各期均较致病组下降，并有统计学差异（P＜0.05）。2．2致病组IL-17在发病初期表达升高，高峰期升高明显（相比有统计学差异，P＜0.05），缓解期下降，但与高峰期相比无统计学差异（P＞0.05）。假干预组各期表达与致病组无差异（P＞0.05）。干预组IL-17表达在发病初期升高，但与致病组没有统计学差异。在发病高峰期升高较致病组明显，并有统计学差异（P＜0.05），在缓解期较致病组表达下降，并有统计学差异（P＜0.05）。2．3致病组IL-6表达在发病初期就已升高，高峰期升高明显，缓解期无明显下降，三时期比较无明显统计学意义（P＞0.05）。假干预组各期表达与致病组无差异（P＞0.05）。干预组IL-6的表达，在发病初期、高峰期和缓解期表达均较相应致病组升高，并有统计学差异（P＜0.05）。结论：1慢性EAE中IL-17表达和疾病严重程度密切相关，而IL-23p19在早期的免疫启动中可能发挥着关键性的作用。2外源性TGF-β1可能通过抑制IL-23p19和促进IL-17的产生对慢性EAE产生双重作用。3 IL-6在慢性EAE病程中发挥着重要的作用。更多还原

【Abstract】 Objective To investigate the changes of IL-23p19, IL-17, IL-6mRNA expression in brains of induced chronic non-remitting model ofexperimental autoimmune encephalomyelitis (EAE), and discuss themechanism of IL-23/Th17cells axis on EAE/MS.Methods 72 adult female C57BL/6 mice were randomly dividedinto the adjuvant -control group, EAE groups, TGF-β1 treatmentgroups, treatment-control groups. MOG35-55 and CFA were injected intosubcutaneous in EAE groups. Animals in TGF-β1 treatment groupsreceived TGF-β1 subcutaneous injection. In addition to immunization,PBS was injected subcutaneously in the treatment-control groups. At theonset, peak and chronic post-immunization, the group mice weresacrificed. The expression of IL-23p19, IL-17, IL-6 mRNA in brainwere detected by using RT-PCR..Results 83.3% mice in EAE groups were developed aprogressive paralysis, which peaked at 20-24 days, and then exhibited achronic disease with a partial clinical recovery. In contrast, 88.9% micein TGF-β1 treatment groups developed the ealier progressive paralysis,which peaked at 17-22 days, and were more prone to recovery. There wereno remarkable difference between EAE and treatment-control groups.The expression of IL-23p19 mRNA of brains in EAE groups increased at early, followed by a descent in the peak phase and chronicphase(p＜0.05) In TGF-β1 treatment groups, The expression ofIL-23p19 mRNA in each phase dramatically deceased (p＜0.05) incontrast to the EAE groups.The expression of IL-17 mRNA of brains in EAE groupsincreased at the peak phase, while the expression of IL-17 mRNA inTGF-β1 increased dramaticly in the TGF-β1 treatment groups(p＜0.05).There was statistically difference in the expression of IL-6 betweenthe EAE group and the TGF-β1 treatment group(p＜0.05).The differencebetween the different phase in the same group was not prominent(p＞0.05)Conclusion1. The expression of IL-17 was correlated with the severity ofdisease,while the expression of IL-23p19 may play an important role inthe initiation of disease.2. Exogenous TGF-β1 may suppress the expression of IL-23p19and promote the expression of IL-17.3. IL-6 may play an important role in the different phase of EAE.更多还原