雷公藤多甙对TLR9 mRNA在实验变态反应性神经炎中表达的影响

Expression of TLR9 mRNA in Experimental Allergic Neuritis and Its Dependence on TWP

观察Toll样受体TLR9在实验性变态反应性神经炎(EAN)大鼠的坐骨神经、脾脏、外周血、淋巴结中的动态表达,以及雷公藤多甙对其的影响,以探讨TLR9与EAN的发病关系,为临床寻找新的治疗自身免疫性疾病的策略奠定理论基础。将68只Lewis大鼠随机分为抗原注射组(EAN组),雷公藤多甙干预组(EAN+TWP组),生理盐水干预对照组(EAN+NS组),完全弗氏佐剂组(CFA组),生理盐水对照组(NS组)。EAN组,EAN+TWP组,EAN+NS组大鼠用周围神经髓鞘蛋白P0180-199+CFA+NS(200μL)免疫,EAN+TWP组从大鼠出现症状开始给予雷公藤多甙灌胃(40mg/(kg·d))干预治疗,观察免疫后大鼠的发病情况和组织病理学改变及TWP对其的影响,并利用RT-PCR检测不同时期TLR9的mRNA表达水平。结果发现,EAN组TLR9mRNA整个实验过程中一直呈上升趋势,前后各时间点相比均有差异(P<0.05),与对照组相比有显著差异。EAN+TWP组大鼠治疗后其症状与EAN+NS组相比稍微改善,坐骨神经炎性细胞浸润减少,EAN+TWP组TLR9的表达在第16天、第24天、第33天均低于EAN+NS组,有显著性差异。由此得出结论:TLR9在EAN的起始及效应阶段具有重要作用。雷公藤多甙可能通过抑制TLR9的表达,减轻EAN发病。

This paper explores the roles of TLR9 in Experimental Autoimmune Neuritis （EAN） and how it is affected by TWP treatment. Male Lewis rats were immunized with the component of PNS myelin sheath protein P0 180-199 （100 mg） and Freund＇s complete adjuvant. The immunized rats were administrated intragastrically with TWP daily till death. The clinical signs of rats and pathological changes in the sciatic nerves of rats were observed. The rats of the immunized group and the control group were sacrificed 7 days, 16 days, 24 days, 33 days after former being immunized. The paraffin sections of embed sciatic nerves were made, which were stained by HE stain and Well＇s stain to observe the histopathology. TLR9 was detected by RT-PCR dynamically, as from spleens, sciatic nerves, peripheral blood and lymphonodes. The results show that the mRNA expression of TLR9 is up-regulated during the whole process of the experiment in EAN group, and is higher than that of the control group. There is a difference of significance between the two neighboring points, with P〈0.05. The expression of TLR9 mRNA in EAN ＋ TWP group is lower than that of EAN ＋ NS group （P〈0.05）. TWP relieves the clinical signs of EAN. In summary, the data show for the first time that TLR9 may play a role in the pathogenesis of EAN in its inducing stage and effecting stage. TWP may ameliorate the EAN through inhabiting the TLR9 activation.