Bergapten alleviates senescence of chondrocytes in osteoarthritis

YE Yuanlan; LIU Kaiwen; PAN Yiming; GUO Fengjin

doi:10.16016/j.2097-0927.202408075

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Bergapten alleviates senescence of chondrocytes in osteoarthritis

Pharmacy | 更新时间：2025-02-26

- Bergapten alleviates senescence of chondrocytes in osteoarthritis
- Journal of Army Medical University Vol. 47, Issue 3, Pages: 262-274(2025)
- 作者机构：
  
  重庆医科大学基础医学院细胞生物学与遗传学教研室，重庆
- 作者简介：
  
  GUO Fengjin， E-mail： guo.fengjin@cqmu.edu.cn
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(82272550)
- DOI：10.16016/j.2097-0927.202408075
  CLC： R285.6;R329.28;R684.3
- Received：19 August 2024，
  
  Revised：24 September 2024，
  
  Published：15 February 2025
- 稿件说明：
移动端阅览
叶元兰，刘恺文，潘一鸣，等.佛手柑内酯延缓骨关节炎软骨细胞衰老［J］.陆军军医大学学报，2025，47（03）：262-274.DOI：10.16016/j.2097-0927.202408075.

YE Y L，LIU K W，PAN Y M，et al.Bergapten alleviates senescence of chondrocytes in osteoarthritis［J］.J Army Med Univ，2025，47（03）：262-274.DOI：10.16016/j.2097-0927.202408075.
叶元兰，刘恺文，潘一鸣，等.佛手柑内酯延缓骨关节炎软骨细胞衰老［J］.陆军军医大学学报，2025，47（03）：262-274.DOI：10.16016/j.2097-0927.202408075. DOI：

YE Y L，LIU K W，PAN Y M，et al.Bergapten alleviates senescence of chondrocytes in osteoarthritis［J］.J Army Med Univ，2025，47（03）：262-274.DOI：10.16016/j.2097-0927.202408075. DOI：

摘要

目的

基于网络药理学、分子对接技术并结合体外实验探讨佛手柑内酯（Bergapten）治疗骨关节炎（osteoarthritis，OA）的作用与机制。

方法

检索Super-Pred、Swiss Target Prediction、Drug Bank等相关数据库获取佛手柑内酯的功能靶点和OA疾病相关作用靶点，将靶点通过Venny 2.1.0进行交集，利用STRING数据库进行蛋白互作（protein-protein interaction，PPI）分析，利用AutoDock Vina软件进行分子对接。在C28/I2细胞中进行体外验证实验，CCK-8检测佛手柑内酯对C28/I2细胞增殖活性的影响，RT-qPCR和细胞免疫荧光检测增殖凋亡、炎症和衰老相关分泌表型（senescence associated secretory phenotype，SASP）因子的表达情况。使用细胞衰老β-半乳糖苷酶（senescence-associated β-galactosidase，SA-β-Gal）染色试剂盒检测SA-β-Gal水平的变化。

结果

筛选得到145个佛手柑内酯治疗OA的潜在作用靶点。PPI分析发现HSP90AA1、EGFR、HIF1A、PIK3CA 4个潜在核心靶点，且分子对接可形成较稳定的复合物。CCK-8发现，10 μmol

L佛手柑内酯处理48 h，C28/I2细胞的存活率最高（

0.05）。细胞免疫荧光、RT-qPCR和Western Blot实验显示，佛手柑内酯处理后，细胞增殖标志物PCNA、CCND1和CCNE1的表达增加（

0.05），而凋亡指标Caspase3和BAX的表达降低（

0.05）。同时，佛手柑内酯处理后，IL-1β诱导的炎性因子IL-6和TNF-α的表达降低，而抗炎因子IL-4和IL-10的表达增加（

0.05）。SASP因子P16、P21

、

MMP9和MMP13的表达也显著减少（

0.05）。Sa-β-Gal染色显示，IL-1β诱导后C28/I2细胞的β-半乳糖苷酶水平显著增加（

0.05），而佛手柑内酯处理后，β-半乳糖苷酶染色的阳性细胞率显著降低（

0.05）。

结论

佛手柑内酯可降低IL-1β刺激的软骨细胞中的SASP因子IL-6、TNF-α、P16、P21

、

MMP9、MMP13等的表达，从而在延缓OA进展中发挥其抗炎、抗衰老的作用。

Abstract

Objective‍

‍To investigate the role and mechanism of bergapten in treating osteoarthritis (OA) based on network pharmacology

molecular docking and in vitro experiments.

Methods‍

‍The functional targets of bergapten and the related targets of OA disease were obtained by searching Super-Pred

Swiss Target Prediction

Drug Bank. The obtained targets were intersected with Venny2.1.0

protein-protein interaction (PPI) analysis was performed using the STRING database

and molecular docking was conducted by AutoDock Vina. Subsequently

human chondrocyte C28/I2 cells were subjected in the following validation experiments. After the cells were treated with 0~50 μmol/L bergapten for 24

48 or 72 h

CCK-8 assay

RT-qPCR and immunofluorescence assay were used to detect the proliferation and apoptosis

and the expression of the molecules related to proliferation

inflammation and senescence (senescence associated secretory phenotype

SASP). Senescence β-galactosidase (SA-β-Gal) staining kit was employed to detect the change of SA-β-Gal level in the cells.

Results‍

There obtained 145 potential targets of bergapten for OA. PPI analysis revealed 4 potential core targets

HSP90AA1

EGFR

HIF1A

and PIK3CA

and they could form relatively stable complex with bergapten. CCK-8 assay showed that 10 μmol/L bergapten treatment for 48 h resulted in the highest proliferative activity of C28/I2 cells (P<0.05). Immunofluorescence assay

RT-qPCR and Western blotting indicated that bergapten treatment induced significant increases in the expression of cell proliferation markers such as PCNA

CCND1 and CCNE1 (P<0.05)

while decreases in the expression of apoptosis markers Caspase3 and BAX (P<0.05). Meanwhile

the expression levels of IL-1β-induced inflammatory factors IL-6 and TNF-α were reduced

while those of anti-inflammatory factors IL-4 and IL-10 were elevated after bergapten treatment (P<0.05). The levels of SASP factors

such as P16

P21

MMP9 and MMP13 were also significantly declined (P<0.05). SA-β-Gal staining displayed that the level of SA-β-Gal in C28/I2 cells was significantly enhanced after IL-1β induction (P<0.05)

while bergapten treatment could decrease the positive cell rate of the staining (P<0.05).

Conclusion‍

Bergapten can reduce the expression of SASP factors such as IL-6

TNF-α

P16

P21

MMP9 and MMP13 in IL-1β-stimulated chondrocytes

and thus exerts its anti-inflammatory and anti-aging effects in delaying the progression of OA.

关键词

Keywords

references

李捷婷 . 基于NMR代谢组学方法的大鼠前交叉韧带损伤后血清代谢特征和相关机制研究［D］. 福州：福建医科大学， 2021 . DOI： 10.27020/d.cnki.gfjyu.2021.000414 http://dx.doi.org/10.27020/d.cnki.gfjyu.2021.000414 .

LI J T . Study on serum metabolic characteristics and related mechanisms after anterior cruciate ligament injury in rats based on NMR metabonomics ［D］. Fuzhou ： Fujian Medical University ， 2021 . DOI： 10.27020/d.cnki.gfjyu.2021.000414 http://dx.doi.org/10.27020/d.cnki.gfjyu.2021.000414 .

MOTTA F ， BARONE E ， SICA A ， et al . Inflammaging and osteoarthritis ［J］. Clin Rev Allergy Immunol ， 2023 ， 64 （ 2 ）： 222 - 238 . DOI： 10.1007/s12016-022-08941-1 http://dx.doi.org/10.1007/s12016-022-08941-1 .

CORYELL P R ， DIEKMAN B O ， LOESER R F . Mechanisms and therapeutic implications of cellular senescence in osteoarthritis ［J］. Nat Rev Rheumatol ， 2021 ， 17 （ 1 ）： 47 - 57 . DOI： 10.1038/s41584-020-00533-7 http://dx.doi.org/10.1038/s41584-020-00533-7 .

BRISOLA A A ， COLOVATI M E S ， CERNACH M C S P ， et al . Association between genetic polymorphisms and osteoarthritis development. Overview of systematic reviews ［J］. Int J Rheum Dis ， 2022 ， 25 （ 7 ）： 733 - 742 . DOI： 10.1111/1756-185X. 14362 http://dx.doi.org/10.1111/1756-185X.14362 .

刘振东 . 甜橙黄酮通过调节SERPINA3延缓膝骨关节炎的作用及机制研究［D］. 扬州：扬州大学， 2023 . DOI： 10.27441/d.cnki.gyzdu.2023.000083 http://dx.doi.org/10.27441/d.cnki.gyzdu.2023.000083 .

LIU Z D . Study on the effect and mechanism of sinensetin in delaying knee osteoarthritis by regulating SERPINA3 ［D］. Yangzhou ： Yangzhou University ， 2023 . DOI： 10.27441/d.cnki.gyzdu.2023.000083 http://dx.doi.org/10.27441/d.cnki.gyzdu.2023.000083 .

RATHBUN A M ， MEHTA R ， RYAN A S ， et al . Duloxetine plus exercise for knee osteoarthritis and depression： a feasibility study ［J］. Osteoarthr Cartil Open ， 2024 ， 6 （ 1 ）： 100426 . DOI： 10.1016/j.ocarto.2023.100426 http://dx.doi.org/10.1016/j.ocarto.2023.100426 .

THORLUND J B ， SIMIC M ， PIHL K ， et al . Similar effects of exercise therapy， nonsteroidal anti-inflammatory drugs， and opioids for knee osteoarthritis pain： a systematic review with network meta-analysis ［J］. J Orthop Sports Phys Ther ， 2022 ， 52 （ 4 ）： 207 - 216 . DOI： 10.2519/jospt.2022.10490 http://dx.doi.org/10.2519/jospt.2022.10490 .

KATZ J N ， ARANT K R ， LOESER R F . Diagnosis and treatment of hip and knee osteoarthritis： a review ［J］. JAMA ， 2021 ， 325 （ 6 ）： 568 - 578 . DOI： 10.1001/jama.2020.22171 http://dx.doi.org/10.1001/jama.2020.22171 .

WANG J ， SUN Y ， LIU J ， et al . Roles of long non-coding RNA in osteoarthritis （Review）［J］. Int J Mol Med ， 2021 ， 48 （ 1 ）： 133 . DOI： 10.3892/ijmm.2021.4966 http://dx.doi.org/10.3892/ijmm.2021.4966 .

谢术欢，冯玛莉 . 佛手柑内酯药理作用研究进展［J］. 海南医学院学报， 2022 ， 28 （ 24 ）： 1917 - 1920 . DOI： 10.13210/j.cnki.jhmu.20210507.003 http://dx.doi.org/10.13210/j.cnki.jhmu.20210507.003 .

XIE S H ， FENG M L . Advances in pharmacological action of bergamot lactone ［J］. J Hainan Med Univ ， 2022 ， 28 （ 24 ）： 1917 - 1920 . DOI： 10.13210/j.cnki.jhmu.20210507.003 http://dx.doi.org/10.13210/j.cnki.jhmu.20210507.003 .

CHEN G P ， XU Q ， DAI M ， et al . Bergapten suppresses RANKL-induced osteoclastogenesis and ovariectomy-induced osteoporosis via suppression of NF-κB and JNK signaling pathways ［J］. Biochem Biophys Res Commun ， 2019 ， 509 （ 2 ）： 329 - 334 . DOI： 10.1016/j.bbrc.2018.12.112 http://dx.doi.org/10.1016/j.bbrc.2018.12.112 .

丁浩南，严佳，唐诗佳，等 . 佛手柑内酯对人牙髓干细胞成骨分化的影响［J］. 南京医科大学学报（自然科学版）， 2023 ， 43 （ 3 ）： 334 - 342 . DOI： 10.7655/NYDXBNS20230306 http://dx.doi.org/10.7655/NYDXBNS20230306 .

DING H N ， YAN J ， TANG S J ， et al . Effects of bergapten on osteogenic differentiation of human dental pulp stem cells ［J］. J Nanjing Med Univ Nat Sci ， 2023 ， 43 （ 3 ）： 334 - 342 . DOI： 10.7655/NYDXBNS20230306 http://dx.doi.org/10.7655/NYDXBNS20230306 .

ZHOU Y ， WANG J ， YANG W D ， et al . Bergapten prevents lipopolysaccharide-induced inflammation in RAW264.7 cells through suppressing JAK/STAT activation and ROS production and increases the survival rate of mice after LPS challenge ［J］. Int Immunopharmacol ， 2017 ， 48 ： 159 - 168 . DOI： 10.1016/j.intimp.2017.04.026 http://dx.doi.org/10.1016/j.intimp.2017.04.026 .

YAO Q ， WU X H ， TAO C ， et al . Osteoarthritis： pathogenic signaling pathways and therapeutic targets ［J］. Signal Transduct Target Ther ， 2023 ， 8 （ 1 ）： 56 . DOI： 10.1038/s41392-023-01330-w http://dx.doi.org/10.1038/s41392-023-01330-w .

钱振昊 . 基于NF-κB及PI 3 K通路探究青藤碱对骨关节炎的作用和机制［D］. 乌鲁木齐：新疆医科大学， 2021 . DOI： 10.27433/d.cnki.gxyku.2021.000909 http://dx.doi.org/10.27433/d.cnki.gxyku.2021.000909 .

QIAN Z H . Exploring the effect and mechanism of sinomenine on osteoarthritis based on NF-κB and PI3K pathways ［D］. Urumqi ： Xinjiang Medical University ， 2021 . DOI： 10.27433/d.cnki.gxyku.2021.000909 http://dx.doi.org/10.27433/d.cnki.gxyku.2021.000909 .

FLANNERY C R ， SEAMAN S A ， BUDDIN K E ， et al . A novel placental tissue biologic， PTP-001， inhibits inflammatory and catabolic responses in vitro and prevents pain and cartilage degeneration in a rat model of osteoarthritis ［J］. Osteoarthr Cartilage ， 2021 ， 29 （ 8 ）： 1203 - 1212 . DOI： 10.1016/j.joca.2021.03.022 http://dx.doi.org/10.1016/j.joca.2021.03.022 .

NEWMAN D J . Modern traditional Chinese medicine： identifying， defining and usage of TCM components ［J］. Adv Pharmacol ， 2020 ， 87 ： 113 - 158 . DOI： 10.1016/bs.apha. 2019.07.001 http://dx.doi.org/10.1016/bs.apha.2019.07.001 .

LIANG Y D ， XIE L ， LIU K ， et al . Bergapten： a review of its pharmacology， pharmacokinetics， and toxicity ［J］. Phytother Res ， 2021 ， 35 （ 11 ）： 6131 - 6147 . DOI： 10.1002/ptr.7221 http://dx.doi.org/10.1002/ptr.7221 .

LATIF K ， KHAN A U ， IZHAR UL HAQUE M ， et al . Bergapten attenuates nitroglycerin-induced migraine headaches through inhibition of oxidative stress and inflammatory mediators ［J］. ACS Chem Neurosci ， 2021 ， 12 （ 18 ）： 3303 - 3313 . DOI： 10.1021/acschemneuro.1c00146 http://dx.doi.org/10.1021/acschemneuro.1c00146 .

PARK S ， BELLO A ， ARAI Y ， et al . Functional duality of chondrocyte hypertrophy and biomedical application trends in osteoarthritis ［J］. Pharmaceutics ， 2021 ， 13 （ 8 ）： 1139 . DOI： 10.3390/pharmaceutics13081139 http://dx.doi.org/10.3390/pharmaceutics13081139 .

GIORGINO R ， ALBANO D ， FUSCO S ， et al . Knee osteoarthritis： epidemiology， pathogenesis， and mesenchymal stem cells： what else is new？ an update ［J］. Int J Mol Sci ， 2023 ， 24 （ 7 ）： 6405 . DOI： 10.3390/ijms24076405 http://dx.doi.org/10.3390/ijms24076405 .

MIAO Y ， CHEN Y W ， XUE F ， et al . Contribution of ferroptosis and GPX4’s dual functions to osteoarthritis progression ［J］. EBioMedicine ， 2022 ， 76 ： 103847 . DOI： 10.1016/j.ebiom.2022.103847 http://dx.doi.org/10.1016/j.ebiom.2022.103847 .

HA S ， KIM H W ， KIM K M ， et al . PAR2-mediated cellular senescence promotes inflammation and fibrosis in aging and chronic kidney disease ［J］. Aging Cell ， 2024 ， 23 （ 8 ）： e14184 . DOI： 10.1111/acel.14184 http://dx.doi.org/10.1111/acel.14184 .

降瑞婵，刘海亮 . 甘草中5种活性物质对人脂肪间充质干细胞的抗衰老作用［J］. 同济大学学报（医学版）， 2021 ， 42 （ 2 ）： 179 - 186 . DOI： 10.12289/j.issn.1008-0392.20291 http://dx.doi.org/10.12289/j.issn.1008-0392.20291 .

JIANG R C ， LIU H L . Anti-aging effect of five active substances from licorice on human adipose-derived stem cells ［J］. J Tongji Univ Med Sci ， 2021 ， 42 （ 2 ）： 179 - 186 . DOI： 10.12289/j.issn.1008-0392.20291 http://dx.doi.org/10.12289/j.issn.1008-0392.20291 .

XIAO X ， WANG W ， LI Y Q ， et al . HSP90AA1-mediated autophagy promotes drug resistance in osteosarcoma ［J］. J Exp Clin Cancer Res ， 2018 ， 37 （ 1 ）： 201 . DOI： 10.1186/s13046-018-0880-6 http://dx.doi.org/10.1186/s13046-018-0880-6 .

WEI Y L ， LUO L J ， GUI T ， et al . Targeting cartilage EGFR pathway for osteoarthritis treatment ［J］. Sci Transl Med ， 2021 ， 13 （ 576 ）： eabb3946 . DOI： 10.1126/scitranslmed.abb3946 http://dx.doi.org/10.1126/scitranslmed.abb3946 .

JIA H R ， MA X Y ， TONG W ， et al . EGFR signaling is critical for maintaining the superficial layer of articular cartilage and preventing osteoarthritis initiation ［J］. Proc Natl Acad Sci U S A ， 2016 ， 113 （ 50 ）： 14360 - 14365 . DOI： 10.1073/pnas.1608938113 http://dx.doi.org/10.1073/pnas.1608938113 .

SUN H ， PENG G X ， CHEN K H ， et al . Identification of EGFR as an essential regulator in chondrocytes ferroptosis of osteoarthritis using bioinformatics， in vivo ， and in vitro study ［J］. Heliyon ， 2023 ， 9 （ 9 ）： e19975 . DOI： 10.1016/j.heliyon.2023.e19975 http://dx.doi.org/10.1016/j.heliyon.2023.e19975 .

KORBECKI J ， SIMIŃSKA D ， GĄSSOWSKA-DOBROWOLSKA M ， et al . Chronic and cycling hypoxia： drivers of cancer chronic inflammation through HIF-1 and NF-κB activation： a review of the molecular mechanisms ［J］. Int J Mol Sci ， 2021 ， 22 （ 19 ）： 10701 . DOI： 10.3390/ijms221910701 http://dx.doi.org/10.3390/ijms221910701 .

XU K ， LU C ， REN X Y ， et al . Overexpression of HIF-1α enhances the protective effect of mitophagy on steroid-induced osteocytes apoptosis ［J］. Environ Toxicol ， 2021 ， 36 （ 11 ）： 2123 - 2137 . DOI： 10.1002/tox.23327 http://dx.doi.org/10.1002/tox.23327 .

PFANDER D ， SWOBODA B ， CRAMER T . The role of HIF-1ɑ in maintaining cartilage homeostasis and during the pathogenesis of osteoarthritis ［J］. Arthritis Res Ther ， 2006 ， 8 （ 1 ）： 104 . DOI： 10.1186/ar1894 http://dx.doi.org/10.1186/ar1894 .

DUVAL E ， BAUGÉ C ， ANDRIAMANALIJAONA R ， et al . Molecular mechanism of hypoxia-induced chondrogenesis and its application in in vivo cartilage tissue engineering ［J］. Biomaterials ， 2012 ， 33 （ 26 ）： 6042 - 6051 . DOI： 10.1016/j.biomaterials.2012.04.061 http://dx.doi.org/10.1016/j.biomaterials.2012.04.061 .

XUE J F ， SHI Z M ， ZOU J ， et al . Inhibition of PI3K/AKT/mTOR signaling pathway promotes autophagy of articular chondrocytes and attenuates inflammatory response in rats with osteoarthritis ［J］. Biomed Pharmacother ， 2017 ， 89 ： 1252 - 1261 . DOI： 10.1016/j.biopha.2017.01.130 http://dx.doi.org/10.1016/j.biopha.2017.01.130 .

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