Zhang Yi, Zhang FengQi, Gao YiNi, Wang MeiJiao, Gao Yan, Li HaiChang, Sun Jing, Wen ChengPing, Xie ZhiJun
School of Basic Medical Science, Zhejiang Chinese Medical University, Hangzhou, China.
The Second School of Clinical Medicine, Zhejiang Chinese Medical University, Hangzhou, China.
Front Pharmacol. 2022 Sep 23;13:952775. doi: 10.3389/fphar.2022.952775. eCollection 2022.
To clarify the mechanism of triptolide (TP) in alleviating the conditions underlying SLE. Eight-week-old MRL/lpr mice were randomly divided into a model group ( = 5), low-dose TP (TP-L) group ( = 5), and high-dose TP (TP-H) group ( = 5). Mice in these groups were gavaged with normal saline, low-dose TP solution, and high-dose TP solution for 8 weeks, respectively. The expression levels of anti-dsDNA, IgG, IgM, IgA, C3, C4, and CREA, BUN, ALT, AST, ALB, and ALP indexes in the serum of mice were detected. The proportion of CD19CD138B220 cells in the spleen and the pathological changes of kidney tissue in the mice were also evaluated. The possible signaling pathways and microRNA (miRNA) targets of TP in the treatment of SLE were analyzed using network pharmacology. The expressions of TLR7 mRNA and miR-146a in Raji cells (a B lymphocyte line) were detected using qPCR before and after intervention with a miR-146a inhibitor. The protein expression levels of TLR7, MyD88, p-IRAK1, and p-NF-κBp65 were detected using western blot analysis. TP could significantly decrease the levels of ds-DNA and IgG, alleviate pathological injury in renal tissue, and upregulate miR-146a expression in the B cells of MRL/lpr mice without obvious liver and kidney toxicity. Network pharmacology analysis showed that TP could mainly regulate the Toll-like receptor signaling pathway, and NF-κB signaling pathway, among others. miRNA target prediction suggested that TP could regulate miRNAs such as miR-146a. cell experiments further confirmed that TP could significantly upregulate miR-146a expression and downregulate the expression of TLR7 mRNA and protein levels TLR7, MyD88, p-IRAK1, and p-NF-κBp65. After intervention with a miR-146a inhibitor, TP had no obvious inhibitory effects on TLR7, MyD88, p-IRAK1, and p-NF-κBp65 expression. TP may exert therapeutic effects on SLE by regulating miR-146a expression, inhibiting the TLR7/NF-κB signaling pathway, and affecting B cell activation.
为阐明雷公藤甲素（TP）缓解系统性红斑狼疮（SLE）潜在病情的机制。将8周龄的MRL/lpr小鼠随机分为模型组（n = 5）、低剂量TP（TP-L）组（n = 5）和高剂量TP（TP-H）组（n = 5）。分别用生理盐水、低剂量TP溶液和高剂量TP溶液对这些组的小鼠进行灌胃8周。检测小鼠血清中抗双链DNA、IgG、IgM、IgA、C3、C4以及肌酐、尿素氮、谷丙转氨酶、谷草转氨酶、白蛋白和碱性磷酸酶指标的表达水平。还评估了小鼠脾脏中CD19⁺CD138⁺B220⁺细胞的比例以及肾脏组织的病理变化。使用网络药理学分析TP治疗SLE可能的信号通路和微小RNA（miRNA）靶点。在用miR-146a抑制剂干预前后，通过qPCR检测Raji细胞（一种B淋巴细胞系）中TLR7 mRNA和miR-146a的表达。使用蛋白质免疫印迹分析检测TLR7、MyD88、p-IRAK1和p-NF-κBp65的蛋白质表达水平。TP可显著降低双链DNA和IgG水平，减轻肾组织病理损伤，并上调MRL/lpr小鼠B细胞中miR-146a的表达，且无明显肝肾毒性。网络药理学分析表明，TP主要可调节Toll样受体信号通路和NF-κB信号通路等。miRNA靶点预测表明，TP可调节如miR-146a等miRNA。细胞实验进一步证实，TP可显著上调miR-146a表达，并下调TLR7 mRNA表达以及TLR7、MyD88、p-IRAK1和p-NF-κBp65的蛋白质水平。在用miR-146a抑制剂干预后，TP对TLR7、MyD88、p-IRAK1和p-NF-κBp65表达无明显抑制作用。TP可能通过调节miR-146a表达、抑制TLR7/NF-κB信号通路以及影响B细胞活化而对SLE发挥治疗作用。
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