狼疮定对系统性红斑狼疮活动期肝肾阴虚证患者外周血单个核细胞中pDC含量及TLR7/TLR9/MyD88通路的影响  

on pDC Content and TLR7/TLR9/MyD88 Pathway in Peripheral Blood Mononuclear Cells of Patients of Systemic Lupus Erythematosus in Active Stage with Syndrome of Liver and Kidney Yin Deficiency

作　　者：汪梅姣[1] 谢志军[1] 王大维[1] 李海昌[1] WANG Meijiao;XIE Zhijun;WANG Dawei;LI Haichang(School of Basic Medicine,Zhejiang University of Chinese Medicine,Hangzhou,310053,China)

机构地区：[1]浙江中医药大学基础医学院,浙江省杭州市310053

出　　处：《中医杂志》2020年第13期1169-1175,共7页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81873266,81973829);国家中医药行业科研专项(201507001-4);国家重点研发计划(2018YFC1705500)。

摘　　要：目的探讨狼疮定治疗系统性红斑狼疮(SLE)活动期肝肾阴虚证的可能作用机制。方法收取SLE活动期肝肾阴虚证患者及正常人全血分离外周血单个核细胞(PBMC),并将SLE活动期肝肾阴虚证患者PBMC分为模型组、狼疮定组(狼疮定冻干粉2μg/μl)、狼疮定+地塞米松组(狼疮定冻干粉2μg/μl+地塞米松注射液10μg/ml)、地塞米松组(地塞米松注射液10μg/ml)。用药24h后收集细胞,采用流式细胞术检测正常人与活动期SLE患者及各组PBMC中浆细胞样树突状细胞(pDC)含量。采用Realtime-PCR法检测各组PBMC细胞Toll样受体7(TLR7)、Toll样受体9(TLR9)、髓样分化因子88(MyD88) mRNA表达水平,Western blotting检测TLR7、TLR9、MyD88、核因子-κB p65(NF-κB p65)蛋白表达,ELISA法检测其上清中干扰素α(IFN-α)含量。结果与正常人比较,SLE活动期患者外周血PBMC中pDC含量减少(P<0.01)。与模型组比较,狼疮定组pDC含量、NF-κB p65蛋白表达增多,MyD88 mRNA及TLR7、TLR9蛋白表达、培养上清中IFN-α含量减少(P<0.05或P<0.01);狼疮定+地塞米松组TLR9、MyD88 mRNA及TLR9、MyD88、NF-κB p65蛋白表达,培养上清中IFN-α含量减少(P<0.05或P<0.01)。与地塞米松组比较,狼疮定组pDC含量增多,TLR7、MyD88 mRNA表达降低,培养上清中IFN-α含量降低;狼疮定+地塞米松组TLR7、TLR9、MyD88 mRNA表达及MyD88、NF-κB p65蛋白表达,培养上清中IFN-α含量均降低(P<0.01)。与狼疮定组比较,狼疮定+地塞米松组NF-κB p65蛋白表达减少(P<0.01)。结论狼疮定及狼疮定联合地塞米松可能通过增多活动期SLE患者PBMC中pDC含量,抑制其PBMC的TLR7/TLR9/MyD88信号通路,进而抑制SLE病情进展,且狼疮定联合地塞米松效果更好。Objective To investigate the possible mechanisms of Langchuangding in the treatment of liver and kidney yin deficiency syndrome in active stage of systemic lupus erythematosus(SLE). Methods Patients with liver and kidney yin deficiency syndrome and normal subject whole blood peripheral blood mononuclear cell(PBMC) of active SLE were collected and divided into a model group, a Langchuangding group(Langchuangding freeze-dry powder 2μg/μl), a Langchuangding+dexamethasone group(Langchuangding freeze-dry powder 2μg/μl + dexamethasone injection 10μg/ml), and a dexamethasone group(dexamethasone injection 10μg/ml). Cells were collected 24 hours after drug administration and flow cytometry was used to detect plasmacytoid dendritic cells(pDC) content in normal and active SLE patients and PBMC in each group. Levels of toll-like receptor 7(TLR7), toll-like receptor 9(TLR9) and myeloid differentiation factor 88(MyD88) mRNA expression in PBMC cells in each group were detected by Realtime-PCR. Protein expression levels of TLR7, TLR9, MyD88 and nuclear factor-κB p65(NF-κB p65) were detected by Western blotting, and the content of IFN-α in the supernatant was detected by ELISA. Results Compared with normal subjects, pDC content in peripheral blood PBMC of active SLE patients decreased(P<0.01). Compared with the model group, pDC content, NF-κB p65 protein expression increased, MyD88 mRNA and TLR7 and TLR9 protein expression decreased, and IFN-α content in culture supernatant decreased in the Langchuangding group(P<0.05 or P<0.01). TLR9, MyD88 mRNA and TLR9, MyD88, NF-κB p65 protein and IFN-α content in culture supernatant was decreased Langchuangding + dexamethasone group(P<0.05 or P<0.01). Compared with dexamethasone group, the content of pDC in Langchuangding group increased, the expression of TLR7 and MyD88 mRNA decreased, and the content of IFN-α in culture supernatant decreased. Expression of TLR7, TLR9, MyD88 mRNA and MyD88, NF-κB p65 protein and IFN-α content in culture supernatant in Langchuangding+dexameth

关 键 词：系统性红斑狼疮 狼疮定 浆细胞样树突状细胞 TOLL样受体7 TOLL样受体9 髓样分化因子88 

分 类 号：R259[医药卫生—中西医结合]