基于网络药理学探讨恒古骨伤愈合剂治疗骨关节炎的机制及动物实验初步验证

邓绍友; 李蓉; 李进涛; 赵玉兰; 王佩锦; 郑红

doi:10.12259/j.issn.2095-610X.S20230701

基于网络药理学探讨恒古骨伤愈合剂治疗骨关节炎的机制及动物实验初步验证

doi: 10.12259/j.issn.2095-610X.S20230701

邓绍友¹,
李蓉²,
李进涛²,
赵玉兰²,
王佩锦^2, ,,
郑红^2, ,

1.
云南省执业药师注册中心，云南昆明　650101
2.
昆明医科大学实验动物学部，云南昆明　650500

基金项目: 云南省科技厅-昆明医科大学应用基础研究联合专项基金重点项目（202201AY070001-032）

详细信息

作者简介:
邓绍友（1964～），男，云南会泽人，理学学士，副主任药师，主要从事药事管理和药理学研究工作

通讯作者:
王佩锦，E-mail：965367067@qq.com

郑红，E-mail：847255170@qq.com

中图分类号: R966
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- 被引次数: 0
出版历程
- 收稿日期: 2023-03-27
- 网络出版日期: 2023-07-17
- 刊出日期: 2023-07-25

Mechanism of Osteoking Improving Osteoarthritis based on Network Pharmacology and DDM Rats

1.
Yunnan Certified Pharmacist Registration Center，Kunming Yunnan 650101
2.
Dept. of Laboratory Animal Science，Kunming Medical University，Kunming Yunnan 650500，China

摘要

摘要: 目的探讨恒古骨伤愈合剂（OK）治疗骨关节炎（OA）的潜在机制并利用DMM大鼠初步验证。方法使用TCMSP数据库收集OK潜在的靶点，运用 GeneCards数据库和OMIM筛选与OA相关的靶基因，利用Cytoscape3.8.2软件建立OK有效成分与OA靶基因间互作的调控网络。使用STRING数据库构建蛋白互作图，进行基因本体（GO）及京都基因与基因组百科全书（KEGG）分析。此外，采用内侧半月板不稳定术（DMM）构建OA大鼠，观察膝关节软骨病变，ELISA检测OK治疗后炎症以及氧化应激水平。结果网络药理学发现，血清白蛋白（ALB），白介素6（IL-6）等80个OK治疗OA的核心基因，主要涉及免疫、炎症反应、氧化应激等生物学过程。动物实验发现，与模型组相比，OK组大鼠关节软骨表面结构趋于正常，软骨细胞脱落少；OK降低血清炎症因子IL-6（P < 0.01）、VEGF（P < 0.01）和氧化应激因子MDA（P < 0.05），改善ALB水平和SOD活性（P < 0.01）。结论 OK可缓解OA软骨组织损伤，改善免疫，抑制炎症和氧化应激反应，初步验证了网络药理学的结果。
- 网络药理学 /
- 恒古骨伤愈合剂 /
- 骨关节炎 /
- DMM大鼠
Abstract: Objective To explore the potential mechanism of Osteoking （OK） in the treatment of Osteoarthritis （OA） and preliminarily verify it with DMM rats. Methods TCMSP database was used to collect potential targets of OK, GeneCards database and OMIM were used to screen target genes related to OA, and Cytoscape 3.8.2 software was used to establish a regulatory network for interaction between OK Active ingredient and OA target genes. The STRING database was used to construct protein interaction maps and GO and KECG were analyzed. In addition, OA rats were constructed using medial meniscus instability surgery （DMM） to observe knee joint cartilage lesions, and ELISA was used to detect inflammation and oxidative stress levels after OK treatment. Results Network pharmacology results showed that 80 core genes were involved in OK therapy for OA, including serum albumin （ALB） and interleukin-6 （IL6）, mainly involving biological processes such as immunity, inflammatory response, oxidative stress, and metabolism. Animal experiments showed that compared with the model group, the surface structure of the articular cartilage in the OK group of rats tended to be normal, with less detachment of chondrocytes; OK reduced serum inflammatory factors IL-6 （P < 0.01）, VEGF （P < 0.01）, and oxidative stress factor MDA （P < 0.05）, improved ALB levels and SOD activity （P < 0.01）. Conclusion OK can alleviate OA cartilage tissue damage, enhance immunity, and inhibit inflammation and oxidative stress reactions, preliminarily verifying the results of network pharmacology.
- Network pharmacology /
- Osteoking /
- Osteoarthritis /
- DMM rats

HTML全文

图 1 OK治疗OA的靶基因

Figure 1. Target genes for OK treatment of OA

下载: 全尺寸图片幻灯片

图 2 “OK-有效成分-靶基因-OA”网络图

Figure 2. Network diagram of "OK-Active Component-Target Gene-OA"

下载: 全尺寸图片幻灯片

图 3 靶点基因 degree 值排序图

Figure 3. Ranking chart of target gene degree values

下载: 全尺寸图片幻灯片

图 4 OK治疗OA的GO -BP富集分析

Figure 4. GO BP enrichment analysis of OK treatment for OA

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图 5 OK治疗OA的KEGG通路分析

Figure 5. KEGG pathway analysis of OK treatment for OA

下载: 全尺寸图片幻灯片

图 6 大鼠关节软骨 HE染色（10×）

A：sham组；B：模型组；C：OK组；黑色箭头表示软骨关节损伤。

Figure 6. HE staining of knee cartilage in rats （10×）

下载: 全尺寸图片幻灯片

图 7 3组大鼠血清免疫与炎症因子

与sham组比较，^**P < 0.01；与DMM组比较，^##P < 0.01。

Figure 7. Serum immunity and inflammatory factors in rats

下载: 全尺寸图片幻灯片

图 8 3组大鼠血清氧化应激因子

与sham组比较，^**P < 0.01；与DMM组比较，^#P < 0.05、^##P < 0.01。

Figure 8. The oxidative stress factors in rats

下载: 全尺寸图片幻灯片

参考文献(24)

[1]	Li M H,Xiao R,Li J B,et al. Regenerative approaches for cartilage repair in the treatment of osteoarthritis[J]. Osteoarthritis and Cartilage,2017,25(10):1577-1587. doi: 10.1016/j.joca.2017.07.004
[2]	Wang P E,Zhang L,Ying J,et al. Bushenhuoxue formula attenuates cartilage degeneration in an osteoarthritic mouse model through TGF-β/MMP13 signaling[J]. Journal of Translational Medicine,2018,16(1):72-86. doi: 10.1186/s12967-018-1437-3
[3]	Wu G,Chen W,Fan H,et al. Duhuo jisheng decoction promotes chondrocyte proliferation through accelerated G1/S transition in osteoarthritis[J]. International Journal of Molecular Medicine,2013,32(5):1001-1013. doi: 10.3892/ijmm.2013.1481
[4]	Liu F,Liu G,Liang W,et al. Duhuo jisheng decoction treatment inhibits the sodium nitroprussiate-induced apoptosis of chondrocytes through the mitochondrial-dependent signaling pathway[J]. International Journal of Molecular Medicine,2014,34(6):1573-1580. doi: 10.3892/ijmm.2014.1962
[5]	吴继昆. 恒古骨伤愈合剂治疗157例骨关节炎疗效总结[J]. 中国现代药物应用,2018,12(24):196-197.
[6]	徐涛涛,郑凌歆,孙岩,等. 恒古骨伤愈合剂治疗膝骨性关节炎的临床疗效及安全性评价[J]. 中国中医骨伤科杂志,2022,30(2):8-12.
[7]	Ru J,Li P,Wang J,et al. TCMSP: A database of systems pharmacology for drug discovery from herbal medicines[J]. Journal of Cheminformatics,2014,6(3):13-25.
[8]	Szklarczyk D,Gable A L,Lyon D,et al. STRING v11: Protein-protein association networks with increased coverage,supporting functional discovery in genome-wide experimental datasets[J]. Nucleic Acids Research,2019,47(1):607-613.
[9]	Culley K L,Dragomir C L,Chang J,et al. Mouse models of osteoarthritis: surgical model of posttraumatic osteoarthritis induced by destabilization of the medial meniscus[J]. Methods in Molecular Biology (Clifton,NJ),2015,1226:143-173.
[10]	王庆甫,马玉峰,殷岳杉. 重新认识膝骨性关节炎的诊断和防治[J]. 中国骨伤,2016,29(9):779-781.
[11]	Lin Z,Fu C,Yan Z,et al. The protective effect of hesperetin in osteoarthritis: An in vitro and in vivo study[J]. Food & Function,2020,11(3):2654-2666.
[12]	Wang S N,Xie G P,Qin C H,et al. Aucubin prevents interleukin-1 beta induced inflammation and cartilage matrix degradation via inhibition of NF-κB signaling pathway in rat articular chondrocytes[J]. International Immunopharmacology,2015,24(2):408-415. doi: 10.1016/j.intimp.2014.12.029
[13]	Wang B W,Jiang Y,Yao Z L,et al. Aucubin protects chondrocytes against iL-1β-Induced apoptosis in vitro and inhibits osteoarthritis in mice model[J]. Drug Design,Development and Therapy,2019,13(9):3529-3538.
[14]	Ling H,Zeng Q,Ge Q,et al. Osteoking decelerates cartilage degeneration in DMM-Induced osteoarthritic mice model through TGF-β/smad-dependent manner[J]. Frontiers in Pharmacology,2021,12(2):678810-678823.
[15]	Kim S,Mcclave S A,Martindale R G,et al. Hypoalbuminemia and clinical outcomes: What is the mechanism behind the relationship?[J]. American Surgeon,2017,83(11):1220-1227. doi: 10.1177/000313481708301123
[16]	Zhang J,Wang T,Fang Y,et al. Clinical significance of serum albumin/globulin ratio in patients with pyogenic liver abscess[J]. Frontiers in Surgery,2021,8(3):677799-677813.
[17]	Nicholson J A,Dowrick A S,Liew S M. Nutritional status and short-term outcome of hip arthroplasty[J]. Journal of Orthopaedic Surgery,2012,20(3):331-345. doi: 10.1177/230949901202000313
[18]	耿秋东,张应生,卢文亚,等. 化湿定痛汤对木瓜蛋白酶诱导的膝骨关节炎大鼠血清IL-1、IL-6、IL-10的影响[J]. 中华中医药杂志,2020,35(9):4654-4657.
[19]	张磊,张劲. 滑膜炎膏治疗膝关节骨性关节炎疗效及对IL-1、IL-6水平影响的临床研究[J]. 湖北中医药大学学报,2020,22(2):41-44.
[20]	李飞龙,谢平金,柴生颋,等. 川芎嗪对膝骨性关节炎大鼠软骨VEGF表达的影响[J]. 中国骨质疏松杂志,2018,24(7):904-909.
[21]	徐慧,陆进明. VEGF在骨关节炎发病机制中的研究进展[J]. 安徽医学,2016,37(7):923-925.
[22]	Pfander D,Kortje D,Zimmermann R,et al. Vascular endothelial growth factor in articular cartilage of healthy and osteoarthritic human knee joints[J]. Annals of the Rheumatic Diseases,2001,60(11):1070-1083. doi: 10.1136/ard.60.11.1070
[23]	Nagao M,Hamilton J L,Kc R,et al. Vascular endothelial growth factor in cartilage development and osteoarthritis[J]. Scientific Reports,2017,7(1):13027-13039. doi: 10.1038/s41598-017-13417-w
[24]	Di Meo S,Venditti P. Evolution of the knowledge of free radicals and other oxidants[J]. Oxidative Medicine and Cellular Longevity,2020,2020:9829176-9829189.

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