A Promising Target of Langchuangding Prescription Treating Systemic Lupus Erythaematosus Integrated Network Pharmacology with HPLC-MS and Molecular Docking.
A Promising Target of Langchuangding Prescription Treating Systemic Lupus Erythaematosus Integrated Network Pharmacology with HPLC-MS and Molecular Docking.
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DOI：
10.31083/j.fbl2711307
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发表时间：
2022-11
期刊：
影响因子：
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通讯作者：
Qianqian Li;Jing Sun;Jifang Tu;Hai-chang Li;Jida Zhang;Huan-peng Gu;Zhijun Xie;Huiqing Lv
中科院分区：
文献类型：
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作者：
Qianqian Li;Jing Sun;Jifang Tu;Hai-chang Li;Jida Zhang;Huan-peng Gu;Zhijun Xie;Huiqing Lv
系统性红斑狼疮（systemic lupus erythematosus，SLE）是一种慢性多系统自身免疫性疾病，可累及几乎所有器官系统，目前尚无有效治疗方法。越来越多的证据表明，活化的T细胞是促进SLE发病的关键因素。传统临床狼疮定方是临床治疗SLE的有效中药方剂，副作用少，患者依从性好。然而，LCD如何影响SLE的机制仍不清楚。方法通过预测LCD和SLE相关靶点，并重叠构建蛋白质相互作用（PPI），筛选核心靶点。用流式细胞仪和Western-blot方法检测LCD血清处理后Jurkat T细胞中目的基因的表达水平。通过HPLC-MS鉴定了LCD中的主要化合物，并与核心目标化合物进行了对接。结果获得LCD中283个蛋白质靶点、1498个SLE靶点和150个常见靶点，构建蛋白质-蛋白质相互作用（PPI）。网络药理学结果提示LCD与CASP 3靶点密切相关。为了验证LCD治疗SLE的药理学机制的预测，我们使用CCK-8试剂盒和流式细胞术分析来研究LCD处理的大鼠血清对β-雌二醇（300 pg/mL）激活的Jurkat T细胞的体外抗增殖作用，然后分析CASP 3表达水平。体外实验证实，LCD血清可通过上调CASP 3表达水平，抑制活化T细胞增殖（p < 0.05），并诱导其凋亡。结合HPLC-MS分析和分子对接，进一步验证了CASP 3靶标与LCD的相互作用。结论LCD对SLE有一定的治疗作用，其机制可能与其上调CASP 3表达水平，诱导活化T细胞凋亡有关。网络药理学和分子对接方法为深化对中医药的认识提供了新的思路。LCD潜在地代表了SLE补充治疗的有前景的治疗处方，没有副作用。
BACKGROUND Systemic lupus erythematosus (SLE) is a chronic multisystem autoimmune disorder affecting almost any organ system without effective treatment. Based on accumulating evidence, activated T cells are key cause promoting the pathogenesis of SLE. A traditional clinic Langchuangding formula (LCD) is an effective clinical traditional Chinese medicine prescription for SLE with few side effects and good patient compliance. However, the mechanism of how LCD affects SLE remains unclear. METHODS Targets related to LCD and SLE were predicted and overlapped to construct protein-protein interaction (PPI) for screening core target. Subsequently, flow cytometry analysis and Western-blot method were used to verify the expression levels of target gene in LCD serum treated-Jurkat T cells. The main compounds of LCD were identified by HPLC-MS and further docked with the core targe. RESULTS 283 protein targets in LCD, 1498 SLE targets and 150 common targets were obtained to construct protein-protein interaction (PPI). Network pharmacology results suggested that LCD was closely related to CASP3 target. To verify the prediction of pharmacological mechanism of LCD treatment for SLE, we investigated the anti-proliferative effects of LCD-treated rat serum on β-oestradiol (300 pg/mL)-activated Jurkat T cells in vitro using a CCK-8 kit and flow cytometry analysis and then analyzed the CASP3 expression levels. Vitro experiments confirmed that LCD serum could suppress the proliferation (p < 0.05) and induce apoptosis of the activated T cells through up-regulating CASP3 expression levels. Interactions between CASP3 target and LCD were further validated integrating HPLC-MS analysis and molecular docking. CONCLUSIONS The results showed that LCD could relieve SLE, which might be attributed to inducing the activated T cells apoptosis by up-regulating CASP3 expression levels. The network pharmacology and molecular docking approach provide a new insight for deepening understanding about TCM. LCD potentially represents a promising therapeutic prescription for SLE supplement treatment with no adverse effects.