大鼠实验性自身免疫性脑脊髓炎模型的建立及丹参酮ⅡA干预对多发性硬化作用的研究  

Establishment of experimental autoimmune encephalomyelitis model in rats and study on the effect of tanshinoneⅡA intervention on multiple sclerosis

作　　者：孙倩 张贺 Sun Qian;Zhang He(Department of Animal Laboratory,General Hospital of Northern Theater Command,Shenyang 110016,China)

机构地区：[1]北部战区总医院实验动物室,沈阳110016

出　　处：《中国临床实用医学》2024年第5期43-48,共6页China Clinical Practical Medicine

基　　金：辽宁省科学技术计划项目(2023JH2/10130076)。

摘　　要：目的建立大鼠实验性自身免疫性脑脊髓炎(EAE)模型,探讨丹参酮ⅡA(TSⅡA)干预对多发性硬化(MS)的作用及可能的作用机制。方法选取SPF级8周龄雄性SD大鼠50只,体质量(250±20)g,体质量范围为230~270 g。采用简单随机分组法将SD大鼠分为对照组、EAE模型组、EAE模型+TSⅡA 1组、EAE模型+TSⅡA 2组、EAE模型+TSⅡA 3组,每组10只。除对照组外,其余四组建立EAE模型,腹腔注射麻醉后双后足垫单点皮下注射0.2 ml抗原佐剂乳化物,尾根部单点皮下注射0.1 ml抗原佐剂乳化物;每只大鼠在腹股沟处皮下注射百日咳毒素溶液500 ng。EAE模型制作当日开始干预,对照组、EAE模型组每日腹腔注射0.9%氯化钠注射液0.5 ml/kg,EAE模型+TSⅡA 1组每日腹腔注射TSⅡA 25 mg/kg,EAE模型+TSⅡA 2组每日腹腔注射TSⅡA 50 mg/kg,EAE模型+TSⅡA 3组每日腹腔注射TSⅡA 100 mg/kg,持续20 d。记录干预期间各组大鼠体质量和Kono评分变化情况。干预结束后,检测血清炎症因子[白细胞介素-2(IL-2)、白细胞介素-23(IL-23)、γ-干扰素(IFN-γ)]水平;根据Elliot公式测量脊髓含水量;尾静脉注射2%伊文思蓝染料观察血脊髓屏障渗透性。结果干预结束后,EAE模型+TSⅡA 2组体质量[(222.51±18.09)g]高于EAE模型组[(152.40±22.98)g]、EAE模型+TSⅡA 1组[(176.20±19.27)g]、EAE模型+TSⅡA 3组[(207.02±17.51)g];EAE模型+TSⅡA 2组Kono评分[(1.48±0.07)分]低于EAE模型组[(4.18±0.14)分]、EAE模型+TSⅡA 1组[(3.41±0.30)分]、EAE模型+TSⅡA 3组[(2.35±0.15)分],差异有统计学意义(P<0.05)。干预结束后,EAE模型+TSⅡA 2组血清IL-2水平[(289.74±34.21)pg/ml]低于EAE模型组[(332.16±20.33)pg/ml]、EAE模型+TSⅡA 1组[(319.72±27.33)pg/ml]、EAE模型+TSⅡA 3组[(310.43±26.32)pg/ml];EAE模型+TSⅡA 2组血清IL-23水平[(82.33±6.24)pg/ml]低于EAE模型组[(96.73±9.32)pg/mll]、EAE模型+TSⅡA 1[(90.62±6.31)pg/ml]、EAE模型+TSⅡA 3组[(84.77±5.39)pg/ml];EAE模型+TSⅡA 2组血�ObjectiveTo establish a model of experimental autoimmune encephalomyelitis(EAE)in rats and investigate the effect of tanshinoneⅡA(TSⅡA)intervention on multiple sclerosis(MS)and its possible mechanism.MethodsA total of 50 SPF grade 8-week-old male SD rats with body mass(250±20)g and body mass ranged from 230 g to 270 g were selected.The SD rats were randomly divided into control group,EAE model group,EAE model+TSⅡA 1 group,EAE model+TSⅡA 2 group,and EAE model+TSⅡA 3 group with 10 rats in each group.Except for the control group,EAE models were established in the other four groups by injecting subcutaneously 0.2 ml antigen adjuvant emulsion into both hind foot pads at single point and 0.1 ml antigen adjuvant emulsion into the tail root at single point after abdominal anesthesia,and in addition,500 ng pertussis toxin solution was injected subcutaneously in the groin.The intervention began on the day the EAE model was made.The control group and EAE model group were intraperitoneally injected 0.9%sodium chloride injection 0.5 ml/kg daily,the EAE model+TSⅡA 1 group was intraperitoneally injected TSⅡA 25 mg/kg daily,the EAE model+TSⅡA 2 group was intraperitoneally injected TSⅡA 50 mg/kg daily,and the EAE model+TSⅡA 3 group was intraperitoneally injected TSⅡA 100 mg/kg daily.The intervention lasted for 20 days.The changes of body mass and Kono score were recorded during the intervention.After the intervention,the levels of serum inflammatory factors[interleukin-2(IL-2),interleukin-23(IL-23),andγ-interferon(IFN-γ)]were detected,the spinal cord water content was measured according to Elliot formula,and the blood-spinal cord barrier permeability was visualized by injecting 2%Evans blue dye into the tail vein.ResultsAfter the intervention,the body mass of EAE model+TSⅡA 2 group[(222.51±18.09)g]was higher than that of EAE model group,EAE model+TSⅡA 1 group,EAE model+TSⅡA 3 group[(152.40±22.98)g,(176.20±19.27)g,(207.02±17.51)g],while the Kono score of EAE model+TSⅡA 2 group[(1.48±0.07)score

关 键 词：多发性硬化 实验性自身免疫性脑脊髓炎 丹参酮ⅡA 动物模型 

分 类 号：R744.51[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]