基于网络药理学和分子对接分析野黄芩苷抗卵泡闭锁的作用机制

doi:10.16431/j.cnki.1671-7236.2023.06.042

摘要/Abstract

摘要： 【目的】基于网络药理学和分子对接方法探讨野黄芩苷抗卵泡闭锁的作用机制，以筛选野黄芩苷抗玉米赤霉烯酮（zearalenone，ZEA）致颗粒细胞损伤的作用途径。【方法】利用PharmMapper、TCMSP、SymMap和GeneCards数据库获得野黄芩苷发挥抗卵泡闭锁作用的靶点，使用Cytoscape v 3.7.2软件构建靶蛋白互作网络关系图并筛选关键靶点；利用DAVID数据库对靶点进行GO功能及KEGG通路富集分析，构建药物-靶点-通路网络图；使用AutoDock Tools 1.5.6软件进行分子对接验证。【结果】共获得34个交集靶点，筛选得到胰岛素样生长因子1受体（insulin-like growth factor 1 receptor，IGF1R）、丝裂原活化激酶3（mitogen-activated protein kinase 3，MAPK3）、血管紧张素原（angiotensinogen，AGT）、G1/S-特异性周期蛋白-D1（G1/S-specific cyclin-D1，CCND1）、纤连蛋白1（fibronectin 1，FN1）、肿瘤坏死因子（tumor necrosis factor，TNF）、半胱天冬酶-3（caspase-3，CASP3）、神经源性基因位点切口同源物蛋白1（neurogenic locus notch homolog protein 1，NOTCH1）和白细胞介素6（interleukin 6，IL6）共9个核心靶点。GO功能富集分析得到基因表达调控、细胞增殖正调控、凋亡过程负调控等264个生物过程条目；线粒体、内质网、细胞质等25个细胞组分条目；生长因子活性、蛋白质结合、细胞外基质结构组成等23个分子功能条目。KEGG通路富集分析得到癌症通路、IL17信号通路、PI3K-Akt信号通路等115条相关通路。分子对接验证结果显示，野黄芩苷与9个核心靶点蛋白的结合能均<0 kJ/mol，说明蛋白可与野黄芩苷自发结合。【结论】野黄芩苷可能通过多靶点、多通路来治疗卵泡闭锁，本研究结果可为阐明野黄芩苷抗ZEA致颗粒细胞损伤的作用机制提供思路。

关键词: 网络药理学; 分子对接; 野黄芩苷; 卵泡闭锁

Abstract: 【Objective】 Based on the method of network pharmacology and molecular docking, the mechanism of scutellarin against follicular atresia was investigated to screen the mechanism of scutellarin against zearalenone (ZEA)-induced granulosa cell damage.【Method】 The databases of PharmMapper, TCMSP, SymMap and GeneCards were used to obtain the target of scutellarin for anti-follicular atresia.The target protein interaction network diagram was constructed using Cytoscape v 3.7.2 software and the key targets were screened.DAVID database was used to analyze GO function and KEGG pathway enrichment of targets, and construct drug-targets-pathway network diagram.AutoDock Tools 1.5.6 software was used for molecular docking verification.【Result】 34 intersection targets were obtained, and 9 core targets were screened, including insulin-like growth factor 1 receptor (IGF1R), mitogen-activated protein kinase 3 (MAPK3), angiotensinogen (AGT), G1/S-specific cyclin-D1 (CCND1), fibronectin 1 (FN1), tumor necrosis factor(TNF), caspase-3 (CASP3), neurogenic locus notch homolog protein 1 (NOTCH1) and interleukin 6 (IL6).GO function enrichment analysis obtained 264 biological processe including regulation of gene expression, positive regulation of cell proliferation, and negative regulation of apoptosis, 25 cellular component including mitochondria, endoplasmic reticulum and cytoplasm, and 23 molecular function including growth factor activity, protein binding and extracellular matrix structure constitute.By KEGG pathway enrichment analysis, 115 related pathways such as cancer pathway, IL17 signaling pathway, PI3K-Akt signaling pathway were obtained.The results of molecular docking verification showed that the affinity of scutellarin with 9 core target proteins was <0 kJ/mol, indicating that the protein could spontaneously bind with scutellarin.【Conclusion】 Scutellarin might antagonize follicular atresia through multiple targets and pathways, which was a candidate research approach for the mechanism of scutellarin's anti ZEA induced granulosa cell injury.

Key words: network pharmacology; molecular docking; scutellarin; follicular atresia

中图分类号:

S859.7

张新越, 孙娜, 孙盼盼, 张华, 范阔海, 尹伟, 孙耀贵, 李宏全. 基于网络药理学和分子对接分析野黄芩苷抗卵泡闭锁的作用机制[J]. 中国畜牧兽医, 2023, 50(6): 2585-2593.

ZHANG Xinyue, SUN Na, SUN Panpan, ZHANG Hua, FAN Kuohai, YIN Wei, SUN Yaogui, LI Hongquan. Mechanism of Scutellarin Against Follicular Atresia Based on Network Pharmacology and Molecular Docking[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2585-2593.

参考文献

[1] KUIPER-GOODMAN T, SCOTT P, WATANABE H.Risk assessment of the mycotoxin zearalenone[J].Regulatory Toxicology and Pharmacology, 1987, 7(3):253-306.
[2] LIU J, APPLEGATE T.Zearalenone (ZEN) in livestock and poultry:Dose, toxicokinetics, toxicity and tstrogenicity[J].Toxins (Basel), 2020, 12(6):377.
[3] ZHU L, YUAN H, GUO C, et al. Zearalenone induces apoptosis and necrosis in porcine granulosa cells via a caspase-3- and caspase-9-dependent mitochondrial signaling pathway[J].Journal of Cellular Physiology, 2012, 227(5):1814-1820.
[4] 赵煜.玉米赤霉烯酮诱导卵巢颗粒细胞凋亡信号通路研究[D].杨凌:西北农林科技大学, 2014. ZHAO Y.Study on zearalenone induced apoptotic pathway of ovary granulosa cells[D].Yangling:Northwest A&F University, 2014.(in Chinese)
[5] 张国梁.赤霉烯酮暴露影响卵巢颗粒细胞发育和卵泡形成的机制研究[D].杨凌:西北农林科技大学, 2017. ZHANG G L.The effects of zearalenone exposure on the ovarian granulosa cell development and follicle assembly[D].Yangling:Northwest A&F University, 2017.(in Chinese)
[6] WANG H, ZHANG Y, ZHANG J, et al.circSLC41A1 resists porcine granulosa cell apoptosis and follicular atresia by promoting SRSF1 through miR-9820-5p sponging[J].International Journal of Molecular Science, 2022, 23(3):1509.
[7] 张颖, 李先强, 李亚娜, 等.颗粒细胞凋亡影响家禽卵泡闭锁的研究进展[J].中国畜牧兽医, 2022, 49(12):444-452. ZHANG Y, LI X Q, LI Y N, et al.Research progress on effects of granulosa cell apoptosis on follicle atresia in poultry[J].China Animal Husbandry & Veterinary Medicine, 2022, 49(12):444-452.(in Chinese)
[8] YAO W, PAN Z, DU X, et al.NORHA, a novel follicular atresia-related lncRNA, promotes porcine granulosa cell apoptosis via the miR-183-96-182 cluster and FoxO1 axis[J].Journal of Animal Science and Biotechnology, 2021, 12(1):103.
[9] LIU J, QI N, XING W, et al.The TGF-β/SMAD signaling pathway prevents follicular atresia by upregulating MORC2[J].International Journal of Molecular Sciences, 2022, 23(18):10657.
[10] HU X, WU X, ZHAO B, et al.Scutellarin protects human retinal pigment epithelial cells against hydrogen peroxide(H₂O₂)-induced oxidative damage[J].Cell and Bioscience, 2019, 9:12.
[11] GAO C, ZHOU Y, JIANG Z, et al.Cytotoxic and chemosensitization effects of scutellarin from traditional Chinese herb Scutellaria altissima L.in human prostate cancer cells[J].Oncology Report, 2017, 38(3):1491-1499.
[12] NIU C W, SHENG Y C, YANG R, et al.Scutellarin protects against the liver injury induced by diosbulbin B in mice and its mechanism[J].Journal of Ethnopharmacology, 2015, 164:301-308.
[13] SUN C Y, NIE J, ZHENG Z L, et al.Renoprotective effect of scutellarin on cisplatin-induced renal injury in mice:Impact on inflammation, apoptosis, and autophagy[J]. Biomed Pharmacother, 2019, 112:108647.
[14] WANG W J, MA X T, HAN J C, et al.Neuroprotective effect of scutellarin on ischemic cerebral injury by down-regulating the expression of angiotensin-converting enzyme and AT1 receptor[J].Public Library of Science, 2016, 11(1):e0146197.
[15] LI Q, CHEN Y J, ZHANG X, et al.Scutellarin attenuates vasospasm through the Erk5-KLF2-eNOS pathway after subarachnoid hemorrhage in rats[J].Journal of Clinical Neuroscience, 2016, 34:264-270.
[16] TIAN X, CHANG L, MA G, et al.Delineation of platelet activation pathway of scutellarin revealed its intracellular target as protein kinase C[J].Biological and Pharmaceutical Bulletin, 2021, 39(2):181-191.
[17] 扆妍妍.野黄芩苷抗ZEA诱导小鼠卵巢颗粒细胞凋亡及其作用机制[D].太谷:山西农业大学, 2020. YI Y Y.The mechanism of scutellarin against ZEA-induced mouse ovarian granulosa cells apoptosis[D].Taigu:Shanxi Agricultural University, 2020.(in Chinese)
[18] SIMONESCHI D, RONA G, ZHOU N, et al.CRL4AMBRA1 is a master regulator of D-type cyclins[J].Nature, 2021, 592(7856):789-793.
[19] 李俊玫.野黄芩苷对人舌鳞癌Tca8113细胞细胞周期及Fas/Fasl介导的细胞凋亡的影响[D].承德:承德医学院, 2013. LI J M.The effects of growth inhibition and apoptosis induction of scutellarin on Tca8113 human tongue cancer cells[D].Chengde:Chengde Medical University, 2013.(in Chinese)
[20] LEE J, KIM E, CHONG K, et al.Atypical induction of HIF-1α expression by pericellular Notch1 signaling suffices for the malignancy of glioblastoma multiforme cells[J].Cell and Molecular Life Science, 2022, 79(10):537.
[21] ZHOU H, CHEN X, CHEN L, et al.Anti-fibrosis effect of scutellarin via inhibition of endothelial-mesenchymal transition on isoprenaline-induced myocardial fibrosis in rats[J]. Molecules, 2014, 19(10):15611-15623.
[22] QIN X, CAO M, LAI F, et al.Oxidative stress induced by zearalenone in porcine granulosa cells and its rescue by curcumin in vitro[J].PLoS One, 2015, 10(6):e0127551.