Uncovering the mechanisms of leech and centipede granules in the treatment of diabetes mellitus-induced erectile dysfunction utilising network pharmacology
Uncovering the mechanisms of leech and centipede granules in the treatment of diabetes mellitus-induced erectile dysfunction utilising network pharmacology
复制标题
利用网络药理学揭示水蛭蜈蚣颗粒治疗糖尿病性勃起功能障碍的机制。
DOI：
10.1016/j.jep.2020.113358
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发表时间：
2021-01-30
影响因子：
5.4
通讯作者：
Chen, Wang Qiang
中科院分区：
文献类型：
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作者：
Ma, Jian Xiong;Wang, Bin;Chen, Wang Qiang
民族药理学相关性：糖尿病引起的勃起功能障碍(DMED)是糖尿病最常见的并发症之一。传统上，水蛭蜈蚣颗粒在东亚各民族医学系统中被用作活血剂，特别是在中国。研究目的：本研究旨在从网络药理学的角度寻找LCG对DMED的潜在基因和作用机制。材料和方法：通过UHPLC-Q-TOF-MS、TCMID和蝙蝠侠-中医药数据库鉴定LCG的活性成分，从DisGeNet、CooLGeN、GeneCards数据库中获取DMed的疾病靶点。在确定LCG的DMED靶点后，构建了蛋白质-蛋白质相互作用(PPI)网络。通过Cytoscape软件的MCODE插件识别HUB基因和重要模块。然后，使用Metascape数据库确定了这些模块的重要信号通路。利用分子操作环境(MOE)对接软件研究了化合物-HUB基因可能的相互作用模式。此外，我们还研究了LCG对链脲佐菌素(STZ)诱导的糖尿病大鼠勃起功能的改善作用及其机制。结果：结合UHPLC-Q-TOF-MS分析和网络药理学研究，筛选出18个活性化合物用于靶向预测。LCG和DMED之间有97个共同的靶基因。KEGG信号通路的丰富主要涉及钙信号通路、核因子-kappaB信号通路、cGMP-PKG信号通路、HIF-1信号通路、PI3K-Akt信号通路和mTOR信号通路。在DMED中，LCG调控了9个HUB基因，包括CXCL8、NOS3、CRH、TH、BDNF、DRD4、ACE、CNR1和HTR1A。分子对接分析结果表明，酪氨酸、熊果酸和L-组氨酸通过产生氢键、H-pi和pi-pi相互作用与相应的HUB基因具有相对稳定的相互作用。值得注意的是，对接结果预测，与酪氨酸、熊果酸和L-组氨酸相比，伐地那非对HUB基因有更高的亲和力。此外，LCG还能提高睾酮水平、勃起频率、ICP/MAP比值、SOD、cGMP、cAMP，降低MDA和AGEs的表达水平。LCG可改善DMED大鼠阴茎组织的组织学改变。因此，LCG可减轻氧化应激，增加NO的产生；在机制探讨中，LCG可显著上调CNR1、NO53、CRH、TH、BDNF和DRD4的mRNA和蛋白表达，而CXCL8、ACE和HTR1A的水平显著高于DMED组。此外，LCG还可激活NO/cGMP/PKG通路、PI3K/Akt/nNOS通路、cAMP/PKA通路，抑制HIF-1α/mTOR通路，从而改善勃起功能。结论：LCG可能通过改变相关代谢途径的基因表达，为DMED的治疗提供新的治疗依据。
Ethnopharmacological relevance: Diabetes mellitus-induced erectile dysfunction (DMED) is one of the most common complications of diabetes mellitus. Leech and centipede granules (LCG) have traditionally been used as blood-activating agents in various ethnomedicinal systems of East Asia, especially in China. It is often used to regulate bodily functions and considered as adjuvant therapy for promoting blood circulation, alleviating blood coagulation, activating meridians, and relieving stasis.Aim of the study: This study aimed to identify potential genes and mechanisms of LCG on DMED from the network pharmacological perspective.Materials and methods: The active components of LCG were identified by UHPLC-Q-TOF-MS, TCMID, and the BATMAN-TCM databases, and the disease targets of DMED were obtained from the DisGeNET, CooLGeN, GeneCards databases. After identifying DMED targets of LCG, a protein-protein interaction (PPI) network was constructed. Hub genes and significant modules were identified via the MCODE plug-in of Cytoscape software. Then, significant signaling pathways of the modules were identified using the Metascape database. The probable interaction mode of compounds-hub genes is examined using Molecular Operating Environment (MOE) docking software. Besides, we investigated the effects and mechanisms of LCG on improving erectile function in the streptozotocin (STZ)-induced diabetic rats model.Results: Combined UHPLC-Q-TOF-MS analysis with network pharmacology study, 18 active compounds were selected for target prediction. There are 97 common target genes between LCG and DMED. Enrichment of the KEGG pathway mainly involves in the calcium signaling pathway, NF-kappa B signaling pathway, cGMP-PKG signaling pathway, HIF-1 signaling pathway, PI3K-Akt signaling pathway, and mTOR signaling pathway. Nine hub genes were regulated by LCG in DMED, including CXCL8, NOS3, CRH, TH, BDNF, DRD4, ACE, CNR1, and HTR1A. The results of molecular docking analysis showed that the tyrosin, ursolic acid, and L-Histidine has a relatively stable interaction with corresponding hub genes via generating hydrogen bonds, H-pi, and pi-pi interactions. Significantly, the results in docking predicted a higher affinity of vardenafil to the hub genes compared to the tyrosin, ursolic acid, and L-Histidine. Furthermore, LCG increased the testosterone, erection frequency, the ratio of ICP and MAP, SOD, cGMP, cAMP as well as decreased the MDA, and AGEs expression levels. And, LCG ameliorated the histological change of penile tissues in DMED rats. Hence, LCG attenuates oxidative stress, increases NO production; For the mechanism exploration, LCG could significantly upregulate the mRNA and protein expression of CNR1, NO53, CRH, TH, BDNF, and DRD4, whereas CXCL8, ACE, and HTR1A levels were significantly higher than those in the DMED group. Moreover, LCG activates the NO/cGMP/PKG pathway, PI3K/Akt/nNOS pathway, cAMP/PKA pathway, and inhibits the HIF-1 alpha/mTOR pathway to improve erectile function.Conclusions: Our results suggest that LCG maybe offer a new therapeutic basis for the treatment of DMED via altering the gene expression of involved metabolic pathways.