基于IRAK1-NF-κB通路探讨解毒祛瘀滋阴方干预MRL/lPr狼疮小鼠机制  

Mechanism of Jiedu Quyu Ziyin Formula on MRL/lpr lupus mice based on IRAK1-NF-κB pathway

作　　者：嵇丽娜 李荣群[1] 包洁[1] 范永升[1] JI Li-na;LI Rong-qun;BAO Jie;FAN Yong-sheng(Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学,杭州310053

出　　处：《中华中医药杂志》2020年第5期2362-2368,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81673863);国家自然科学基金青年科学基金项目(No.81803980);浙江省自然科学基金青年项目(NO.LQ18H270004)。

摘　　要：目的:研究解毒祛瘀滋阴方对MRL/lpr狼疮小鼠及其脾细胞和CD4+T细胞白细胞介素-1受体相关激酶(IRAK1)信号通路异常活化的影响,探讨解毒祛瘀滋阴方对IRAK1-NF-κB信号通路的作用,为其治疗系统性红斑狼疮(SLE)临床用药提供良好的理论支持。方法:制备解毒祛瘀滋阴方冻干粉,HPLC法检测其有效成分及含量;分离来自MRL/lpr狼疮小鼠的脾脏细胞和CD4+T细胞,流式检测CD4+T细胞的纯度;以LPS诱导细胞,采用CCK8法检测LPS和中药对细胞的活力。将MRL/lpr狼疮小鼠随机分为西药组、中药组和模型组,MRL/Mp小鼠作为对照组,药物干预结束后,HE染色观察肾脏病理改变;RT-qPCR和Western blot技术检测小鼠脾细胞、CD4+T细胞和脾脏IRAK1及下游NF-κB信号通路之间的表达水平。结果:解毒祛瘀滋阴方中检测到有效成分没食子酸、芍药苷、阿魏酸和异阿魏酸;中药组MRL/lpr小鼠的狼疮症状减轻,脾脏IRAK1-NF-κB信号通路的表达水平显著降低(P<0.05)。解毒祛瘀滋阴方冻干粉以100、10、1μg/mL为最佳给药浓度,可降低LPS诱导的IRAK1及其下游蛋白的磷酸化,并抑制IFN-α和IFN-β的表达(P<0.05)。结论:解毒祛瘀滋阴方对MRL/lpr小鼠的狼疮症状有治疗作用,并能通过下调MRL/lpr小鼠及其CD4+T细胞中的IRAK1-NF-κB信号通路,抑制LPS诱导的细胞活化,解毒祛瘀滋阴方治疗SLE作用机制可能是通过调控IRAK1信号通路来实现的。Objective: To study the effects of Jiedu Quyu Ziyin Formula on the abnormal activation of IRAK1 signaling pathway in MRL/lpr lupus mice and their spleen cells and CD4+T cells, the effect of Jiedu Quyu Ziyin Decoction on IRAK1-NF-κB signaling pathway can be explored to provide good theoretical support for its clinical use in the treatment of systemiclupus erythematosus(SLE). Methods: The Jiedu Quyu Ziyin Formula powder was prepared, and the effective components and contents in the Jiedu Quyu Ziyin Formula were determined by HPLC. The spleen cells and CD4+T cells from MRL/lpr lupus mice were isolated, and the purity of CD4+T cells was detected by flow cytometry. Cells were induced, and the viability of LPS and Chinese medicine on the cells was detected by the CCK8 method. MRL/lpr lupus mice were randomly divided into western medicine group, traditional Chinese medicine group and model group. MRL/Mp mice were used as control group. After drug intervention, HE staining was used to observe renal pathological changes. Expression levels between mouse spleen cells, CD4+T cells, and spleen IRAK1 and downstream NF-κB signaling pathways were detected by RT-qRCR and Western blot. Results: The active ingredients gallic acid, paeoniflorin, ferulic acid and isoferulic acid were detected in Jiedu Quyu Ziyin Formula. The lupus symptoms of MRL/lpr mice in the traditional Chinese medicine group were alleviated, and the expression level of IRAK1-NF-κB signaling pathway in the spleen significantly reduced(P<0.05). Jiedu Quyu Ziyin Formula(100, 10, 1μg/mL) as the optimal concentrations, could reduce the phosphorylation of IRAK1 and its downstream proteins induced by LPS, and inhibit the IFN-α and IFNexpression(P<0.05). Conclusion: Jiedu Quyu Ziyin Formula has therapeutic effects on lupus symptoms in MRL/lpr mice, and can inhibit LPS-induced cells by down-regulating the IRAK1-NF-κB signaling pathway in MRL/lpr mice and their CD4+T cells. The mechanism of Jiedu Quyu Ziyin Formula in treating SLE may be achieved by regulating IRAK1

关 键 词：解毒祛瘀滋阴方 系统性红斑狼疮 CD4+T细胞 白细胞介素-1受体相关激酶 核转录因子ΚB MRL/lpr狼疮小鼠 CCK8法 机制 

分 类 号：R285.5[医药卫生—中药学]