Network pharmacology and molecular docking to study the mechanism of action of alpinumisoflavone in a tem-poromandibular joint osteoarthritis cell model
10.12016/j.issn.2096-1456.2024.08.002
- VernacularTitle:基于网络药理学和分子对接研究高山金莲花素对颞下颌关节骨关节炎细胞模型的作用机制
- Author:
Zejie WANG
1
;
Gaoyi WU
Author Information
1. 佳木斯大学口腔医学院,黑龙江省口腔生物医学材料及临床应用重点实验室,黑龙江 佳木斯(154000)
- Keywords:
temporomandibular joint;
osteoarthritis;
temporomandibular joint osteoarthritis cell model;
alpinu-misoflavone;
network pharmacology;
molecular docking;
flavonoids;
mandibular condylar chondrocytes;
apoptosis;
extracellular matrix degradation;
interleukin 1β;
B
- From:
Journal of Prevention and Treatment for Stomatological Diseases
2024;32(8):578-588
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the potential role of alpinumisoflavone(AIF)in the treatment of temporomandibu-lar joint osteoarthritis(TMJOA)cell model through network pharmacology and molecular docking and to provide a re-search basis for AIF in the treatment of TMJOA.Methods GeneCards,OMIM,DisGeNET,and PharmGKB databases were used to screen TMJOA disease targets,and PharmMapper and HERB were used to retrieve AIF-related targets.The intersection targets of the compounds and diseases were uploaded to the STRING database to obtain the key targets for GO and KEGG enrichment analysis,while the key targets in related signaling pathways were evaluated through mo-lecular docking.Approval was obtained from the Ethics Committee to extract condylar chondrocytes from 3-week-old SD rats.The CCK-8 assay was used to detect AIF cytotoxicity on condylar chondrocytes.Condylar chondrocytes were in-duced with 10 ng/mL interleukin 1β(IL-1β)for 24 h to construct a TMJOA cell model.The experiment was divided in-to three groups:control group,comprising condylar chondrocytes cultured in DMEM for 48 h;IL-1β group,comprising condylar chondrocytes pre-cultured in DMEM for 24 h,after which IL-1β was added to the original culture medium to obtain a medium concentration of 10 ng/mL and allowed to culture for 24 h;and the IL-1β+10 μmol/L AIF group,com-prising condylar chondrocytes pre-cultured in DMEM medium containing 10 μmol/L AIF for 24 h,after which IL-1β was added to the original culture medium to obtain a medium concentration of 10 ng/mL and allowed to culture for 24 h.The effect of AIF on condylar chondrocyte apoptosis in the TMJOA cell model was detected by flow cytometry.The experiment was divided into four groups:control group,IL-1β group,IL-1β+10 μmol/L AIF group,and IL-1β+30 μmol/L AIF group.The IL-1β+30 μmol/L AIF group was pre-cultured in DMEM containing 30 μmol/L AIF for 24 h,after which IL-1β was added to the original culture medium to obtain a medium concentration of 10 ng/mL and allowed to culture for 24 h.The remaining three groups were cultured in the same manner as before.The mRNA and protein ex-pression of apoptosis-associated B-cell leukemia/lymphoma-2(Bcl2),cysteinyl aspartate specific protease 3(caspase-3),matrix degradation-associated a disintegrin and metalloproteinase with thrombospondin motifs 4(ADAMTS4),matrix me-talloproteinase 3(MMP3),and matrix metalloproteinase 13(MMP13)were detected by qPCR and western blot,by AIF in the TMJOA cell model.Results The PharmMapper and HERB database search yielded 300 AIF compound targets.The GeneCards,OMIM,DisGeNET,and PharmGKB databases yielded 378 TMJOA disease targets.Thirty-three poten-tial common targets were obtained by intersecting compounds with disease targets.The common targets were uploaded into the STRING database to obtain 31 key targets that were mainly associated with apoptosis and extracellular matrix degradation.This process may be associated with the MAPK,estrogen,and TNF signaling pathways.The molecular docking results showed that AIF has good binding activity with extracellular signal-regulated kinase 1/2(ERK1/2)and estrogen receptor gene 1/2(ESR1/2),which are key targets in the MAPK and estrogen signaling pathways.The CCK-8 assay showed that AIF had no obvious cytotoxicity to condylar chondrocytes.The cell experiments showed that AIF inhib-ited apoptosis in the IL-1β+10 μmol/L AIF group compared to the IL-1β group.Compared to the IL-1β group in the IL-1β+10 μmol/L AIF group and the IL-1β+30 μmol/L AIF group,AIF upregulated Bcl2 and downregulated caspase-3 mRNA and protein expression and inhibited ADAMTS4,MMP3,and MMP13 mRNA and protein expression.Conclusion AIF inhibited apoptosis in the TMJOA cell model by upregulating Bcl2 and downregulating caspase-3 mRNA and pro-tein expression,and inhibited extracellular matrix degradation induced by IL-1β,thereby delaying TMJOA progression.
