狼疮定通过调节TLR7-IRF7-IFNα通路治疗系统性红斑狼疮的机制
The mechanism of Langchuangding in treatment of systemic lupus erythematosus via modulating TLR7-IRF7-IFNα pathway.
作者信息
Wang Meijiao, Zhang Yiyang, Zhai Yingqi, Li Haichang, Xie Zhijun, Wen Chengping
机构信息
Research Institute of Chinese Medicine Clinical Foundation and Immunology, School of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, Zhejiang, China.
出版信息
Heliyon. 2024 Feb 17;10(5):e26022. doi: 10.1016/j.heliyon.2024.e26022. eCollection 2024 Mar 15.
OBJECT
Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by aberrant activity of the immune system. Plasmacytoid dendritic cells (pDCs) which the main producer of activated type I interferon, are related to SLE disease activity. To investigate the mechanism of Langchuangding (LCD) improving SLE based on TLR7-IRF7-IFNα pathway.
METHODS
SLE patients were randomly divided into Chinese medicine combined with western medicine (CWM) group and western medicine (WM) group, to observe the effect of LCD. The percent of pDCs in peripheral blood of SLE patients were detected by flow cytometry, and the influence of LCD on gene expression in SLE patients were detected by gene microarray. Mouse bone marrow cells were differentiated into dendritic like cells (DLC), then divided into Blank, immune complex (IC), LCD and dexamethasone (DXM) group. Employed RT-qPCR to detect , and mRNA, and western blotting to determinate TLR7, MyD88, and p-IRF7 proteins. The IFNα in SLE patients were detected by enzyme-linked immunosorbent assay (ELISA). Employ dual luciferase to observe the interferon stimulated response element (ISRE) gene.
RESULTS
pDCs in WM group was higher than that of CWM group. The plasma IFNα in CWM group was significantly lower than that in WM group. The gene microarray showed that the gene expression of IFNα related signaling pathway in peripheral blood mononuclear cell (PBMC) and genes related to activation and proliferation of immune cells were down-regulated after LCD treatment. The DLCs , and mRNA were down-regulated, TLR7, MyD88, and p-IRF7 proteins were significantly reduced, and the supernatant IFNα was significantly decreased in LCD group. LCD were mildly inhibited activation of ISRE in 293T cells.
CONCLUSIONS
In certain degree, LCD is beneficial to SLE patients. LCD therapy SLE may be through TLR7 signaling pathway, and IRF7 may be a promising therapeutic target for the treatment of SLE.
目的
系统性红斑狼疮（SLE）是一种复杂的自身免疫性疾病，其特征为免疫系统异常激活。浆细胞样树突状细胞（pDCs）是活化的I型干扰素的主要产生者，与SLE疾病活动相关。旨在探讨狼疮定（LCD）基于TLR7-IRF7-IFNα通路改善SLE的机制。
方法
将SLE患者随机分为中西医结合（CWM）组和西药（WM）组，观察LCD的疗效。采用流式细胞术检测SLE患者外周血中pDCs的百分比，采用基因芯片检测LCD对SLE患者基因表达的影响。将小鼠骨髓细胞分化为树突状样细胞（DLC），然后分为空白、免疫复合物（IC）、LCD和地塞米松（DXM）组。采用RT-qPCR检测 、 和 mRNA，采用蛋白质印迹法测定TLR7、MyD88和p-IRF7蛋白。采用酶联免疫吸附测定（ELISA）检测SLE患者体内的IFNα。采用双荧光素酶观察干扰素刺激反应元件（ISRE）基因。
结果
WM组的pDCs高于CWM组。CWM组的血浆IFNα显著低于WM组。基因芯片显示，LCD治疗后外周血单个核细胞（PBMC）中IFNα相关信号通路的基因表达以及与免疫细胞活化和增殖相关的基因表达下调。LCD组DLCs的 、 和 mRNA下调，TLR7、MyD88和p-IRF7蛋白显著减少，上清液IFNα显著降低。LCD轻度抑制293T细胞中ISRE的激活。
结论
在一定程度上，LCD对SLE患者有益。LCD治疗SLE可能通过TLR7信号通路，且IRF7可能是治疗SLE的一个有前景的治疗靶点。
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