龙胆泻肝汤对实验性自身免疫性葡萄膜炎大鼠M1/M2巨噬细胞极化平衡的调控作用  

Regulatory role of Longdan Xiegan Decoction in polarization balance of M1/M2 macrophages in rats with experimental autoimmune uveitis

作　　者：殷学伟 郭励劼 周梦贤 邱岩 毕宏生[4,5] 郭大东[5] YIN Xuewei;GUO Lijie;ZHOU Mengxian;QIU Yan;BI Hongsheng;GUO Dadong(Shandong University of Traditional Chinese Medicine,Jinan 250014,Shandong Province,China;Capital Medical University,Beijing 100069,China;The Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250002,Shandong Province,China;Affiliated Eye Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250002,Shandong Province,China;Shandong Provincial Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Therapy of Ocular Diseases,Key Laboratory of Integrated Traditional Chinese and Western Medicine Eye Disease Prevention and Treatment Technology(Enhanced)in Universities of Shandong Province,Eye Institute of Shandong University of Traditional Chinese Medicine,Jinan 250002,Shandong Province,China)

机构地区：[1]山东中医药大学,山东省济南市250014 [2]首都医科大学,北京市100069 [3]山东中医药大学第二附属医院,山东省济南市250002 [4]山东中医药大学附属眼科医院,山东省济南市250002 [5]山东省中西医结合眼病防治重点实验室,山东省高校中西医结合眼病防治技术(强化)重点实验室,山东中医药大学眼科研究所,山东省济南市250002

出　　处：《眼科新进展》2021年第5期408-412,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金项目(编号:81873163);山东省自然科学基金重点项目(编号:ZR2020KC024);山东中医药科技发展计划项目(编号:2015-145)。

摘　　要：目的探讨龙胆泻肝汤(Longdan Xiegan decoction,LXD)对实验性自身免疫性葡萄膜炎(EAU)大鼠M1/M2巨噬细胞极化平衡的调控作用。方法将48只雌性Lewis大鼠随机分为正常对照组、EAU模型组和LXD干预组,其中EAU模型组和LXD干预组大鼠首先诱导并建立EAU模型,LXD干预组大鼠每天给予LXD灌胃处理12 d,EAU组大鼠给予相同体积的PBS缓冲液灌胃处理12 d。实时荧光定量PCR(Q-PCR)和酶联免疫吸附实验(ELISA)检测免疫后12 d三组大鼠脾脏、淋巴结及眼组织中iNOS和Arg1 mRNA及蛋白的表达;此外,采用流式细胞仪检测以上各组织中M1型、M2型巨噬细胞极化水平,分析LXD对EAU大鼠M1/M2巨噬细胞极化平衡的影响。结果免疫后12 d,与正常对照组相比,EAU模型组大鼠各组织中iNOS mRNA和蛋白表达均升高,而Arg1 mRNA和蛋白表达均降低(均为P<0.05);与EAU模型组相比,LXD干预组大鼠脾脏、淋巴结以及眼组织中iNOS表达均降低,而Arg1表达均升高(均为P<0.05)。流式细胞仪检测结果显示,免疫后12 d,EAU模型组大鼠各组织中M1型巨噬细胞极化水平升高,而M2型巨噬细胞极化水平降低(均为P<0.05),呈现不均衡表达;与EAU模型组相比,LXD干预组大鼠各组织中M1型巨噬细胞极化水平下降,而M2型巨噬细胞极化水平升高,两者比例逐渐恢复均衡。结论在EAU大鼠中,M1/M2巨噬细胞极化比例紊乱,LXD可明显降低M1型巨噬细胞极化水平以及相关因子iNOS表达,提高M2型巨噬细胞极化水平以及相关因子Arg1表达,从而发挥对EAU大鼠M1/M2巨噬细胞极化平衡的调控作用。Objective To investigate the regulatory role of Longdan Xiegan Decoction(LXD)in the polarization balance of M1/M2 macrophages in rats with experimental autoimmune uveitis(EAU).Methods The female Lewis rats were randomly divided into normal control(NC)group,EAU model group and LXD intervention group.First,the EAU model was induced in rats in EAU model group and LXD intervention group.After immunization,rats in the LXD intervention group were given LXD intragastrically every day for 12 days,while the rats in EAU group were treated with the same volume of sterilized phosphate buffered saline(PBS)intragastrically every day for 12 days.The expression of iNOS and Arg1 mRNA and protein levels in spleen,lymph node and eye tissue were detected by quantitative polymerase chain reaction(Q-PCR)and enzyme-linked immunosorbent assay(ELISA),respectively;in addition,the expression levels of the polarization of M1 and M2 macrophages cells were detected by flow cytometry,and the effect of LXD on the polarization balance of M1/M2 macrophages in EAU rats was further investigated.Results Compared with the NC group,the mRNA and protein levels of iNOS in spleen,lymph node and eye tissue in EAU model group were increased,whereas the mRNA and protein levels of Arg1 were decreased 12 days after immunization(all P<0.05).Compared with EAU model group,the expression levels of iNOS in LXD group were significantly decreased,and the expression levels of Arg1 in spleen,lymph node and eye tissue were significantly elevated(all P>0.05).The results of flow cytometry showed that 12 days after immunization,the polarization level of M1 macrophages in the tissues of the EAU model group increased,while the polarization level of M2 macrophages decreased(all P<0.05),suggesting that the proportion was in disorder.In addition,compared with EAU model group,the level of M1 macrophages in LXD group was significantly decreased,whereas the level of M2 macrophages was significantly increased(all P>0.05),and the proportion of M1/M2 macrophages gradually returned t

关 键 词：龙胆泻肝汤 实验性自身免疫性葡萄膜炎 INOS Arg1 M1/M2巨噬细胞极化 

分 类 号：R773[医药卫生—眼科]