D-青霉胺诱导的自身免疫性疾病模型大鼠脾组织中微RNA的表达  

Expression of microRNAs in spleens of autoimmune disease model rats induced by D-penicillamine

作　　者：任禹珂 姜华[1] 张頔[1] 杨艳伟[1] 王超[1] 张河战[1] 李波[1] 林志[1] REN Yu-ke;JIANG Hua;ZHANG Di;YANG Yan-wei;WANG Chao;ZHANG He-zhan;LI Bo;LIN Zhi(National Institutes for Food and Drug Control National Center for Safety Evaluation of Drugs,Key Laboratory of Beijing for Nonclinical Safety Evaluation of Drugs,Beijing 100176,China)

机构地区：[1]中国食品药品检定研究院,国家药物安全评价监测中心,药物非临床研究北京市重点实验室,北京100176

出　　处：《中国药理学与毒理学杂志》2021年第5期353-359,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：国家科技重大专项(2018ZX09201017-001)。

摘　　要：目的建立不同剂量D-青霉胺诱导的Brown Norway(BN)大鼠自身免疫性疾病模型,通过RT-qPCR检测大鼠脾组织中4种微RNA(miRNA)miR-126a-5p,miR-146a-5p,miR-27a-3p和miR-155-5p表达的变化,探讨miRNA作为生物标志物预测药源性自身免疫毒性的意义。方法采用30只BN雄性大鼠,分为溶媒对照组(生理盐水)、D-青霉胺150和450 mg·kg^(-1)组,每组10只,分别在给药7和14 d后(各5只)解剖,ELISA检测血清抗核抗体(ANA);测体重,计算脾系数;HE染色观察脾组织病理改变;使用茎环法RT-qPCR检测脾组织中miRNA表达水平,并绘制受试者操作特征曲线(ROC),比较ANA,miR-126a-5p,miR-146a-5p,miR-27a-3p和miR-155-5p对药源性自身免疫毒性的预测价值。结果与溶媒对照组相比,D-青霉胺给药组大鼠脾系数增加(P<0.05),脾组织生发中心活跃伴易染体巨噬细胞数目增多,血清ANA水平增高(P<0.05),脾组织中miR-126a-5p,miR-146a-5p,miR-27a-3p和miR-155-5p表达水平均显著升高(P<0.01)。ANA,miR-126a-5p,miR-146a-5p,miR-27a-3p和miR-155-5p的ROC曲线下面积(AUC)分别为0.783(95%CI,0.618~0.947),0.887(95%CI,0.732~1.000),0.851(95%CI,0.705~0.997),0.846(95%CI,0.701~0.991)和0.828(95%CI,0.667~0.989)。结论D-青霉胺诱导的自身免疫性疾病模型大鼠脾组织miRNA出现异常表达,并且与传统自身免疫生物标志物ANA相比具有更高的预测价值,提示miRNA有可能成为早期诊断药源性自身免疫毒性的生物标志物。OBJECTIVE To establish an autoimmune disease model of Brown Norway(BN)rats induced by different doses of D-penicillamine,detect the expression changes of four types of miRNA(miR-126 a-5 p,miR-155-5 p,miR-146 a-5 p and miR-27 a-3 p)in spleen tissues of rats by RT-qPCR,and to explore the significance of mi RNA as novel biomarkers for predicting drug induced autoimmune toxicity.METHODS Thirty BN rats were randomly divided into the solvent control group(normal saline),D-penicillamine 150 and 450 mg·kg^(-1) groups with 10 rats each.These rats were sacrificed after 7 and 14 d of administration(5 rats per time point)respectively,and their serum was separated for anti-nuclear antibody(ANA)test.The mass of the spleen was measured and histopathological examination was performed.The expression of miRNA in spleen tissue was detected by stem loop RT-qPCR,and the receiver operating curve(ROC)was drawn.Using the pathological change of the spleen as the standard,the predictive values of ANA,miR-126 a-5 p,miR-146 a-5 p,miR-27 a-3 p and miR-155-5 p for drug-induced autoimmunity were compared.RESULTS Compared with the solvent control group,the spleen coefficient of D-penicillamine groups was increased(P<0.05).Histopathological examination showed that the germinal center was active with the increase of the number of tingible body macrophages,and the level of ANA in serum was increased(P<0.05).Stem-loop RT-q PCR results confirmed that the expression levels of the four kinds of miRNA(miR-126 a-5 p,miR-146 a-5 p,miR-27 a-3 p,miR-155-5 p)in spleen tissue were significantly up-regulated(P<0.01).The areas under the ROC curve(AUC)of ANA,miR-126 a-5 p,miR-146 a-5 p,miR-27 a-3 p,miR-155-5 p were 0.783(95%CI:0.618-0.947),0.887(95%CI:0.732-1.000),0.851(95%CI:0.705-0.997),0.846(95%CI:0.701-0.991)and 0.828(95%CI:0.667-0.989)respectively.CONCLUSION The expression of mi RNA is aberrant in the autoimmune disease animal model induced by D-penicillamine in BN rats,and these miRNAs have higher predictive value than the traditional autoimmune marker A

关 键 词：D-青霉胺 自身免疫性疾病 微RNA 

分 类 号：R392.3[医药卫生—免疫学]