【摘要】 目的:探讨雷公藤内酯醇(triptolide,Tri)治疗实验性自身免疫性脑脊髓炎(EAE)的有效性和可能机制。方法:雌性C57BL/6小鼠随机分为EAE组(28只)、Tri治疗组(20只)和佐剂组(18只)。EAE组和Tri治疗组用髓鞘少突胶质细胞糖蛋白35-55多肽(MOG35-55)免疫建立EAE模型,佐剂组以等量生理盐水代替。Tri治疗组于免疫后5d开始按100μg/(kg.d)腹腔注射Tri,EAE组和佐剂组等量的生理盐水作为对照。观察3组小鼠的发病情况以及病理变化,免疫后18～20d无菌分离脾淋巴细胞,分2组,分别为对照组(只加200μL的细胞悬液)、实验组(细胞悬液中加入MOG35-55每孔终浓度为10mg/L),并将各组细胞悬液接种于96孔板中置于37℃、50mL/LCO2条件下孵育48h。用MTT法检测MOG35-55刺激前后各组脾淋巴细胞的增殖情况,ELISA检测各组细胞培养上清液中细胞因子INF-γ、IL-17、IL-10和IL-4的水平。结果:Tri能够显著改善EAE的发病,减轻炎性浸润,并且可抑制MOG35-55诱导的脾淋巴细胞增殖反应(P<0.01)及INF-γ的分泌(P<0.05),促进IL-10的分泌(P<0.05),而对IL-17和IL-4的分泌没有显著性影响。结论:Tri可以有效抑制小鼠EAE,这种抑制可能与其对脾细胞中INF-γ表达的下调及对IL-10分泌的上调作用密切相关。更多还原

【Abstract】 AIM:To explore the therapeutic effectivity and the possible mechanism of triptolide(Tri)on experimental autoimmune encephalomyelitis(EAE).METHODS:All female C57BL/6 mice were randomly divided into EAE group(28),Tri treated group(20)and adjuvant group(18).Mice in EAE and treated groups were immunized with myelin oligodendrocyte glycoprotein peptides 35-55(MOG35-55),adjuvant group was injected at the same time,but instead of MOG35-55 with normal saline.Tri was intraperitoneally injected in the dosage of 100 μg/(kg·d)in treated group on day 5 post-immunization(p.i.),and mice in EAE and adjuvant group injected with normal saline as control.The clinical feature and pathological changes were observed and the splenic lymphocytes were prepared on days 18-20 p.i.The cell cultures were divided into the control group(only 200 μL of cell suspension)and the experimental group(cell suspension in the presence of 10 mg/L MOG35-55).Then all of them were inoculated in 96-well flat-bottom plates under 37℃,50 mL/L CO2.After 48 h,the proliferation assay was determined by MTT,and the supernatants were harvested for the detection of INF-γ,IL-17,IL-10 and IL-4 by ELISA.RESULTS:Tri treatment showed an significantly protective action on EAE.After the intervention of Tri,the levels of IL-10 were increased(P<0.05),but the secretion of INF-γ and proliferation response of splenic lymphocytes induced by MOG35-55 were statistically significantly inhibited(P<0.05 and P<0.01,respectively).There were no influences on the amount of IL-17 and IL-4 by Tri.CONCLUSION:Tri is an effective drug in suppressing murine EAE.This suppression is surposed to be related to downregulation of INF-γ and upregulation of IL-10 secretion in splenic lymphocytes.更多还原