枯芩与条芩提取物对湿热型溃疡性结肠炎大鼠Th17/Treg免疫失衡的调节机制研究  

Mechanisms of extracts of Pith-nodecayed and Pith-decayed of Scutellariae Radix against Th17/Treg immune imbalance in damp-heat ulcerative colitis rats

作　　者：黄伟[1,2] 罗雅琴[3] 于新阳[3] 董红敬 王晓 HUANG Wei;LUO Yaqin;YU Xinyang;DONG Hongjing;WANG Xiao(Shandong Academy of Chinese Medicine,Institute of Chinese Materia Medica Pharmacology(Pharmacological Experimental Center),Jinan Shandong 250014,China;Qilu University of Technology(Shandong Academy of Sciences),Shandong Analysis and Test Center,Jinan Shandong 250014,China;Affiliated Hospital of Shandong University of Chinese Medicine(First Clinical College),Jinan Shandong 250014,China)

机构地区：[1]山东省中医药研究院中药药理研究所药理实验中心,山东济南250014 [2]齐鲁工业大学(山东省科学院)山东省分析测试中心,山东济南250014 [3]山东中医药大学附属医院第一临床学院,山东济南250014

出　　处：《中国药物警戒》2024年第5期535-539,546,共6页Chinese Journal of Pharmacovigilance

基　　金：中央本级重大增减支项目“名贵中药资源可持续利用能力建设项目”(2060302);国家自然科学基金资助项目(81704029);山东省自然科学基金联合基金重点资助项目(ZR 2022LZY007);山东省中医药高层次人才培育项目(鲁卫人才字[2023] 37号);齐鲁卫生与健康杰出青年人才资助项目(鲁卫人才字[2020] 3号);山东省中医药科技项目(Z-2022098T、M-2022027)。

摘　　要：目的比较枯芩、条芩提取物对湿热型溃疡性结肠炎(ulcerative colitis,UC)大鼠模型辅助性T细胞17(Th17)/调节性T细胞(Treg)细胞因子及转录因子维甲酸相关孤儿受体(ROR-γt)/叉头翼状螺旋转录因子3(Foxp3)表达的影响。方法50只健康SD大鼠随机分为5组:正常组、模型组、枯芩组、条芩组和美莎拉嗪组,每组10只。采用高脂高糖饲料喂养+高度白酒灌胃+5%葡聚糖硫酸钠(dextran sulfate sodium,DSS)联合诱导法,建立湿热型UC大鼠模型;各组分别于造模第1天开始灌胃给药,枯芩组、条芩组给药剂量均为5.25 g·kg^(-1)·d^(-1),美沙拉嗪组给药剂量为0.266 g·kg^(-1)·d^(-1),正常组、模型组灌服等体积生理盐水。连续给药28 d。给药结束后,麻醉大鼠,腹主动脉取血,分离血清,Elisa法检测大鼠血清白介素-6(IL-6)、IL-17A、IL-10、转化生长因子-β_(1)(TGF-β_(1))水平;分离脾脏和结肠组织,采用流式细胞术检测脾脏Th17、Treg细胞比例,计算Th17/Treg比值;采用实时荧光定量聚合酶链式反应(qRT-PCR)法检测结肠组织ROR-γt、Foxp3 mRNA表达水平;免疫蛋白印迹(western blot)法检测结肠组织ROR-γt、Foxp3蛋白表达水平。结果与正常组比较,模型组大鼠血清IL-6、IL-17A、脾脏Th17细胞比例、Th17/Treg、结肠组织ROR-γt mRNA和蛋白表达水平均显著增加(P<0.01),血清IL-10、IL-35、TGF-β_(1)、脾脏Treg细胞比例、结肠组织Foxp3 mRNA和蛋白表达水平均显著降低(P<0.01);给药28 d后,与模型组比较,各给药组大鼠血清IL-6、IL-17A、脾脏Th17细胞比例、Th17/Treg、结肠组织ROR-γt mRNA和蛋白表达水平出现不同程度的降低(P<0.05或P<0.01),血清IL-10、IL-35、TGF-β_(1)、脾脏Treg细胞比例、结肠组织Foxp3 mRNA和蛋白表达水平出现不同程度的升高(P<0.05或P<0.01);与枯芩组比较,条芩组、美沙拉嗪组在降低IL-6、IL-17A,增加IL-10、IL-35、TGF-β_(1)水平,调节ROR-γt mRNA和蛋白表达方面(P<0.05或P<0.01)�Objective To compare the effects of extracts from Pith-decayed and Pith-nodecayed products of Scutellariae Radix on expressions of helper T cell 17 (Th17)/regulatory T cell (Treg) cytokines and the transcription factor ROR-γt/Foxp3 in rats with damp-heat ulcerative colitis (UC).Methods Fifty healthy SD rats were randomly divided into five groups:the normal group,model group,Pith-decayed products of Scutellariae Radix group,Pith-nodecayed products of Scutellariae Radix group and mesalazine group,with 10rats in each.A damp-heat UC rat model was established via the combination of high-fat and high-sugar diet+high-liquor administration+5%dextran sodium sulfate (DSS).Each group was given intragastric administration on the first day of modeling,respectively.The dosage was 5.25 g·kg^(-1)·d^(-1) for the Pith-decayed products group and Pith-nodecayed products group,but 0.266 g·kg^(-1)·d^(-1) for the mesalazine group.The normal group and model group were given an equal volume of normal saline.The drug was administered for 28 consecutive days.After administration,the rats were anesthetized,blood was collected from the abdominal aorta,serum was separated,and serum levels of interleukin-6(IL-6),IL-17A,IL-10 and transforming growth factor-β_(1)(TGF-β_(1)) were detected by Elisa.Spleen and colon tissues were separated,the proportion of Th17 and Treg cells in spleens was detected by flow cytometry,and the Th17/Treg ratio was calculated.Real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression levels of ROR-γt and Foxp3 in colon tissues.The expression levels of ROR-γt and Foxp3 in colon tissues were detected by Western blot.Results Compared with the normal group,serum levels of IL-6 and IL-17A,the proportion of Th17 cells in spleens,Th17/Treg,and mRNA and protein expression levels of ROR-γt in colon tissues of the model group were significantly increased(P<0.01) while serum levels of IL-10,IL-35,TGF-β_(1),the proportion of Treg cells in spleens and the expression of F

关 键 词：黄芩 枯芩 条芩(或子芩) 提取物 湿热型溃疡性结肠炎 Th17/Treg失衡 细胞因子 ROR-γt Foxp3 大鼠 

分 类 号：R977[医药卫生—药品]