Jieduquyuziyin Prescription-Treated Rat Serum Suppresses Activation of Peritoneal Macrophages in MRL/Lpr Lupus Mice by Inhibiting IRAK1 Signaling Pathway
Jieduquyuziyin Prescription-Treated Rat Serum Suppresses Activation of Peritoneal Macrophages in MRL/Lpr Lupus Mice by Inhibiting IRAK1 Signaling Pathway
复制标题
解毒祛瘀滋饮方大鼠血清通过抑制IRAK1信号通路抑制MRL/Lpr狼疮小鼠腹膜巨噬细胞的活化
DOI：
10.1155/2019/2357217
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发表时间：
2019-11-03
影响因子：
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通讯作者：
Li, Rongqun
中科院分区：
文献类型：
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作者：
Ji, Lina;Hou, Xiaoli;Li, Rongqun
系统性红斑狼疮(SLE)是一种慢性自身免疫性疾病，解毒祛瘀子饮方是中国多年来批准的治疗SLE有效的中药方剂。然而，它在治疗这种疾病中的作用机制在很大程度上是未知的。本研究旨在探讨JP大鼠血清能否通过下调IRAK1信号通路，抑制MRL/LPR小鼠腹腔巨噬细胞的活化，从而达到改善SLE的作用。制备JP处理的大鼠血清，体外分离MRL/LPR狼疮小鼠的巨噬细胞，用CCK8法检测JP对细胞活力的影响。脂多糖诱导和shRNA慢病毒感染后，免疫荧光染色检测JP对细胞IRAK1表达的影响。用双抗体夹心法测定细胞培养上清液中肿瘤坏死因子-α和白介素6的含量。免疫印迹法检测iRAK1、p-iRAK1、TRAF6、iKbα、p-ikBα、IKK + ikk、NF-κB、p-NF-κB蛋白的表达水平。我们研究了JP在MRL/LPR小鼠腹腔巨噬细胞中的作用，并确定了可能的作用机制。结果表明，JP可降低脂多糖诱导的IRAK1及其下游蛋白的磷酸化，抑制炎症细胞因子的表达，包括肿瘤坏死因子-α和IL-6。此外，将shRNA慢病毒导入细胞后，JP检测结果趋于一致。结论：JP可能通过下调IRAK1-NF-κB信号通路抑制小鼠腹腔巨噬细胞的活化，IRAK1可能是JP治疗SLE的潜在靶点。
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease, and Jieduquyuziyin prescription (JP) is a traditional Chinese medicine (TCM) formula that has been testified to be effective for SLE treatment as an approved hospital prescription for many years in China. However, its mechanism of action in the treatment of this disease is largely unknown. The purpose of this study was to determine whether JP-treated rat serum can inhibit the activation of peritoneal macrophages in MRL/lpr mice by downregulating the IRAK1 signaling pathway, thereby achieving the effect of improving SLE. The JP-treated rat serum was prepared, and the peritoneal macrophages of MRL/lpr lupus mice were isolated in vitro, and the effect of JP on cell viability was detected by the CCK8 method. After LPS induction and shRNA lentiviral transfection, the effect of JP on the expression of IRAK1 in cells was detected by immunofluorescence staining. The content of TNF-α and IL-6 in the cell supernatant was determined by ELISA. The expression of IRAK1, NF-κB, TNF-α, and IL-6 mRNA was detected by RT-PCR, and the expression levels of IRAK1, p-IRAK1, TRAF6, IKBα, p-IKBα, IKK + IKK, NF-κB, and p-NF-κB proteins was detected by western blot method. We investigated the role of JP in peritoneal macrophages of the MRL/lpr mouse and identified the possible mechanisms of action. The results showed that JP could reduce the phosphorylation of IRAK1 and its downstream proteins induced by LPS and inhibit the expression of inflammatory cytokines, including TNF-α and IL-6. In addition, after the transfection of cells with shRNA lentiviral, the results of JP tended to be consistent. In conclusion, JP may inhibit the activation of peritoneal macrophages in MRL/lpr mice by downregulating the IRAK1-NF-κB signaling pathway, and IRAK1 may be a potential target for JP treatment of SLE.