首页分享基于网络药理学的女贞子抗骨质疏松作用机制研究

基于网络药理学的女贞子抗骨质疏松作用机制研究

来源：花匠小妙招时间：2025-05-12 16:22

目的基于网络药理学相关技术分析中药女贞子防治骨质疏松的主要活性成分及相关靶点通路,探讨其防治骨质疏松的潜在作用机制。方法采用TCMSP和TCMID数据库收集女贞子中活性成分,通过Swiss Target Prediction、Drugbank 以及IPA 对活性成分作用靶点进行预测;同时采用TTD、Drugbank 以及IPA 对骨质疏松疾病靶点进行预测,最后进行映射得到女贞子有效成分-疾病靶点。运用Cytoscape3.2.1软件构建“活性成分-靶点”网络图,进行可视化分析及ClueGO富集分析,此外借助在线分析工具DAVID 和 KOBAS 进行KEGG通路富集分析,筛选出显著富集的通路(P＜0.05),最后采用小鼠原代细胞观察女贞子醇提物对破骨细胞分化的影响。结果通过数据库收集得到女贞子化学成分共119个,其中根据OB≥30%、DL≥0.18以及文献报道的有效成分共17个,包括木犀草素(luteolin),槲皮素(quercetin),熊果酸(ursolic acid),花旗松素(taxifolin),β-谷甾醇(beta-sitosterol)等;同时收集骨质疏松作用靶点161个,映射后靶点为33个,其中包含与骨生成相关靶点ALP,RUNX2,CA2,ESR1,ESR2,PPARγ等。ClueGO富集分析结果显示,其生物途径主要包含类固醇代谢调节,平滑肌细胞增生,破骨细胞分化,组织重构,生殖发育等;KEGG分析结果显示与炎症,破骨分化,细胞凋亡等信号通路相关。细胞实验结果显示,女贞子醇提物1 μg·mL-1具有显著减低破骨细胞分化的作用(P＜0.05)。结论女贞子醇提物具有抑制破骨细胞分化的作用,其防治骨质疏松的机制可能是与抑制破骨细胞的分化,抗炎,抗脂肪生成及促骨生成相关,并通过AMPK, TNF,NF-κB,PPARγ等信号通路发挥作用。

Abstract

OBJECTIVE To analyze the major active components and related target pathways of Ligustrum lucidum for the treatment of osteoporosis based on network pharmacology, and explore its potential mechanism for prevention and treatment of osteoporosis. METHODS The active components of Ligustrum lucidum were collected using TCMSP and TCMID databases and the targets of active components were predicted by Swiss Target Prediction, Drugbank and IPA. In addition, the targets of osteoporosis were predicted by TTD, Drugbank and IPA. Finally, the two parts of the results were mapped to obtain the targets of active component for treating osteoporosis. The Cytoscape3.2.1 software was used to construct the “component-target network” for visual analysis and ClueGO enrichment analysis. In addition, the online analysis tools DAVID and KOBAS were used to perform KEGG pathway enrichment analysis to screen out significantly enriched pathways (P<0.05). Finally, the primary osteoclast of mouse bone marrow was cultured to observe the effect of ethanol extract of Ligustrum lucidum on osteoclast differentiation. RESULTS A total of 119 components of Ligustrum lucidum were collected from the database, and 17 active ingredients were screened according to OB≥30% and DL≥0.18, including luteolin, quercetin, ursolic acid, taxifolin and beta-sitosterol, etc. In addition, 161 targets on osteoporosis were collected and among which 33 targets were mapped, including bone generation related targets ALP, RUNX2, CA2, ESR1, ESR2, PPARγ and so on. ClueGO enrichment analysis showed that its biological pathways mainly included steroid metabolism regulation, smooth muscle cell proliferation, osteoclast differentiation, tissue remodeling as well as reproductive development. KEGG analysis depicted its association with signaling pathways such as inflammation, osteoclast differentiation and apoptosis etc. The results of cell experiments showed that the ethanol extract of Ligustrum lucidum of 1 μg·mL-1 significantly decreased the differentiation of osteoclasts (P<0.05). CONCLUSION The ethanol extract of Ligustrum lucidum has the effect of inhibiting the differentiation of osteoclasts. The mechanism for prevention and treatment of osteoporosis may be related to the reduction of osteoclast differentiation, anti-inflammation, anti-lipogenesis and osteogenic growth, and the signaling of AMPK, TNF, NF-κB, PPARγ may also plays significant roles.

关键词

网络药理学 /女贞子 /骨质疏松 /靶点 /机制{{custom_keyword}} /

Key words

network pharmacology /Ligustrum lucidum /osteoporosis /target /mechanism{{custom_keyword}} /

覃小燕, 韩晓玲, 张媛, 韦秋, 李洁, 毛浩萍.基于网络药理学的女贞子抗骨质疏松作用机制研究[J]. 中国药学杂志, 2021, 56(5): 368-378 https://doi.org/10.11669/cpj.2021.05.005

QIN Xiao-yan, HAN Xiao-ling, ZHANG Yuan, WEI Qiu, LI Jie, MAO Hao-ping.Mechanism of Ligustrum lucidum in Treatment of Osteoporosis Based on Network Pharmacology[J]. Chinese Pharmaceutical Journal, 2021, 56(5): 368-378 https://doi.org/10.11669/cpj.2021.05.005

参考文献

[1] Chinese Medical Association. Guidelines for diagnosis and treatment of primary osteoporosis (2017)[J]. Chin J Osteoporos Bone Miner Res(中华骨质疏松和骨矿盐疾病杂志), 2017, 10(5):413-444.
[2] ZHANG H, HE J W, WANG C, et al. Associations of polymorphisms in the SOST gene and bone mineral density in postmenopausal Chinese women[J]. Osteoporos Int, 2014, 25(12):2797-2803.
[3] GAFFNEY-STOMBERG E, INSOGAN K L, RODRIGUEZ N R, et al. Increasing dietary protein requirements in elderly people for optimal muscle and bone health [J]. J Am Geriatr Soc, 2009, 57(6):1073-1079.
[4] LISTED N. Assessment of fracture risk and its application to screening for postmenopausal osteoporosis. Report of a WHO Study Group [M].(WHO)World Health Organ Tech Rep Ser, 1994, 843: 1-129.
[5] ERKUS S L, TURGUT A L, KOSE O. Remodelling of humeral fracture in a child with osteopetrosis tarda following conservative management--a 7-year follow-up: a case report[J]. Malays Orthop J, 2019, 13(2): 42-44.
[6] WANG J. Treatment and prevention of senile osteoporosis[J]. Soc Sci Theory(科教导刊), 2018,(8): 146-147.
[7] XIAO Y P, ZENG J, JIAO L N, et al. Review for treatment effect and signaling pathway regulation of kidneytonifying traditional Chinese medicine on osteoporosis[J]. China J Chin Mater Med(中国中药杂志), 2018, 43(1):21-30.
[8] XU S N, ZHUANG L, ZHAI Y Y, et al. The material basis and mechanism of prevention and treatment of osteoporosis based on network pharmacology[J]. Chin Pharm J(中国药学杂志), 2018, 53 (22): 1913-1920.
[9] ZHU W T, FAN X M, WEI H, et al. Mechanism research of apatinib-treated breast cancer based on network pharmacology[J]. Chin Pharm J(中国药学杂志), 2014, 49(16):3148-3151.
[10] RU J L. Construction and Utilization of Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platfrom [D]. Yangling:Northwest A & F University, 2015.
[11] HOPKINS A L. Network pharmacology[J]. Nat Biotechnol, 2007, 25(10): 1110-1111.
[12] LI S, ZHNAG B. Traditional Chinese medicine network pharmacology: theory, methodology and application [J]. Chin J Nat Med(中华医学杂志), 2013, 11(2): 110-120.
[13] YANG K L, WANG Y, AI L, et al. Research ideas on Tongmai Yangxin Prescription in treatment of coronary artery disease based on network pharmacology[J]. Chin Tradit Herb Drugs(中草药), 2015, 46(20):2979-2984.
[14] QIU Y M, WEN G, JJIE L I, et al. Progress on lipidomics analytical methods and their applications in studies of traditional Chinese medicines[J]. China J Chin Mater Med(中国中药杂志), 2019, 44(9): 1760-1766.
[15] LISTED N. NIH consensus development panel on osteoporosis prevention, diagnosis, and therapy, march 7-29, 2000: highlights of the conference [J]. South Med J, 2001, 94(6):569-573.
[16] KANIS J A, MCCLOSKEY E V, HARVEY N C, et al. Intervention thresholds and the diagnosis of osteoporosis[J]. J Bone Miner Res, 2015, 30(10):1747-1753.
[17] WARD L M, RAUCH F. Anabolic therapy for the treatment of osteoporosis in childhood [J]. Curr Osteoporos Rep, 2018, 16(3):269-276.
[18] WHAYNE T F. Hypertriglyceridemia: an infrequent, difficult-to-predict, severe metabolic and vascular problem associated with estrogen administration[J]. Curr Vasc Pharmacol, 2020,18(3):254-261.
[19] LIN X S, WANG H Y, ZHANG Z, et al. Effects of acupoint application therapy with Tiangui powder on osteoporosis in ovariectomized rats through TGF-beta1 and Smad2/3 signaling pathway[J]. Orthop Surg, 2019, 11(1): 143-150.
[20] TABATAEI-MALAZY O, SALARI P, KHASHAYAR P, et al. New horizons in treatmen of osteoporosis [J]. Daru, 2017, 25(1):2.
[21] KIM I R, KIM S E, BAEK H S, et al.The role of kaempferolinduced autophagy on differentiation and mineralization of osteoblastic MC3T3-E1 cells[J]. BMC Complement Altern Med, 2016, 16 (1):333.
[22] MELGUIZO-RODRIGUEZ L, MANZANO-MORENO F J, ILLESCAS-MONTES, et al. Bone protective effect of extra-virgin olive oil phenolic compounds by modulating osteoblast gene expression [J]. Nutrients, 2019, 11(8):1722.
[23] PANG X G, CONG Y, BAO N R, et al. Quercetin stimulates bone marrow mesenchymal stem cell differentiation through an estrogen receptor-mediated pathway[J]. Biomed Res Int, 2018, 2018: 4178021.
[24] ZHANG M M. Estrogen and estrogen receptors on bone metabolism regulation[J]. Chin J Osteoporos(中国骨质疏松杂志), 2019, 25(5):704-708.
[25] TAHARA K, KOISO H, OSAKI Y H, et al. Malignancy-associated hypercalcemia related with receptor activator of NF-κB ligand (RANKL) expression in T-cell acute lymphoblastic leukemia[J]. Acta Haematol, 2019, 141(3):135-137.
[26] WALY N E, REFAIY A, ABOREHAB N M. IL-10 and TGF-β: roles in chondroprotective effects of glucosamine in experimental osteoarthritis [J]. Pathophysiol, 2017, 24(1): 45-49.
[27] FRANCESCONE R, HOU V, GRIVENNIKOV S I. Microbiome, inflammation, and cancer[J]. Cancer J, 2014, 20(3):181-189.
[28] NISHIKAWA A, ISHIDA T, TAKETSUNA M, et al. Safety and effectiveness of daily teriparatide in a prospective observational study in patients with osteoporosis at high risk of fracture in Japan:final report[J]. Clin Interv Aging, 2016, 11:913-925.
[29] KIM J, LEE Y J, KIM J M, et al. PPARgamma agonists induce adipocyte differentiation by modulating the expression of lipin-1, which acts as aPPAR gamma phosphatase[J]. Int J Biochem Cell Biol, 2016, 81:57-66.