Exploration of the mechanism of Bushen Bitong Prescription in the treatment of knee osteoarthritis based on network pharmacology and experimental verification
XIANG Wenyuan1 YI Lin1 DENG Yingjie2 FANG Rui2▲
1.The Fourth School of Clinical Medicine, Xinjiang Medical University, Xinjiang Uygur Autonomous Region, Urumqi 830054, China;
2.the Second Department of Orthopaedics, Hospital of Traditional Chinese Medicine, Xinjiang Medical University, Xinjiang Uygur Autonomous Region, Urumqi 830099, China
Abstract：Objective To explore the mechanism of Bushen Bitong Prescription in the treatment of knee osteoarthritis (KOA) by network pharmacology and experimental verification. Methods Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform were used to obtain the active ingredients and targets of Bushen Bitong Prescription. GeneCards, OMIM, PharmGkb, TTD, and DrugBank databases were used to search KOA disease related targets, and the intersection of drug and disease targets was selected. The obtained active ingredients and related targets were introduced into Cytoscape software, and the network map of “Bushen Bitong Prescription - active ingredients-KOA targets” was drawn to screen the key active ingredients. Potential targets were imported into the STRING database to plot the protein-protein interaction network. The core targets of Bushen Bitong Prescription in the treatment of KOA were screened, and GO enrichment and KEGG annotation were analyzed. Human chondrocytes were selected and set up in control group, injury group, and administration group. The control group was cultured normally, the injury group and administration group were cultured in the medium containing interleukin (IL)-1β (10 ng/ml) DMEM /F12, and the administration group was treated with 0.444 7 μg/ml Bushen Bitong Prescription liquid. After 24 h of culture, the cell morphology of the three groups was observed. The levels of inflammatory factors were detected by enzyme-linked immunosorbent assay, and the expression of cartilage repair factors in the three groups was detected by qRT-PCR and Western blot. Results A total of 186 active ingredients of Bushen Bitong Prescription were screened, mainly including quercetin, luteolin, kaempferol, etc., mainly through the key targets of TNF, MMP1, CXCL2, IL1B, and signaling pathways of PI3K/Akt, T cell receptors, toll-like receptors, hypoxy-inducer factor-1, TNF, IL-17, and others. It involved, oxidative stress, apoptosis, inflammation, and other biological processes. Microscopically, the number of cells decreased in the injury group and increased in the administration group. The levels of tumor necrosis factor-α (TNF-α), IL-1β, and IL-6 in injury group were higher than those in control group, and the levels of transforming growth factor-β (TGF-β), insulin-like growth factor-1 (IGF-1), basic fibroblast growth factor (BFGF), and the gene and protein expression levels of SOX9 and COL2A1 were lower than those in control group (P＜0.05). The levels of TNF-α, IL-1β, and IL-6 in administration group were lower than those in injury group, and the levels of TGF-β, IGF-1, BFGF, and the gene and protein expression levels of SOX9 and COL2A1 were higher than those in injury group (P＜0.05). Conclusion Bushen Bitong Prescription may promote the repair of injured chondrocytes by relieving inflammatory cell infiltration and participating in the regulation of SOX9 and COL2A1.
向文远1 易林1 邓迎杰2 方锐2▲. 基于网络药理学和实验验证探讨补肾痹通方治疗膝骨关节炎的作用机制[J]. 中国医药导报, 2023, 20(15): 27-34.
XIANG Wenyuan1 YI Lin1 DENG Yingjie2 FANG Rui2▲. Exploration of the mechanism of Bushen Bitong Prescription in the treatment of knee osteoarthritis based on network pharmacology and experimental verification. 中国医药导报, 2023, 20(15): 27-34.
[1] Charlier E，Deroyer C，Ciregia F，et al. Chondrocyte dedifferentiation and osteoarthritis（OA） [J]. Biochem Pharmacol，2019，165：49-65.
[2] Safiri S，Kolahi AA，Smith E，et al. Global，regional and national burden of osteoarthritis 1990-2017：a systematic analysis of the Global Burden of Disease Study 2017 [J]. Ann Rheum Dis，2020，79（6）：819-828.
[3] Cross M，Smith E，Hoy D，et al. The global burden of hip and knee osteoarthritis：estimates from the global burden of disease 2010 study [J]. Ann Rheum Dis，2014，73（7）：1323-1330.
[4] 中华医学会骨科学分会关节外科学组，中国医师协会骨科医师分会骨关节炎学组，国家老年疾病临床医学研究中心（湘雅医院），等.中国骨关节炎诊疗指南（2021年版）[J].中华骨科杂志，2021，41（18）：1291-1314.
[5] 方锐，孟庆才，王拥军，等.补肾通络方对大鼠膝关节炎软骨中半胱氨酸天冬氨酸特异性蛋白酶3的调节[J].中国临床康复，2006，10（43）：67-69，229.
[6] 方锐，孟庆才，邓迎杰，等.膝骨关节炎患者血清和关节液中基质金属蛋白酶1，3、基质金属蛋白酶抑制剂1及白细胞介素1β含量与补肾通络方的干预[J].中国组织工程研究与临床康复，2008，336（28）：5581-5585.
[7] Ru J，Li P，Wang J，et al. TCMSP：a database of systems pharmacology for drug discovery from herbal medicines [J]. J Cheminform，2014，6：13.
[8] Zhang JL，Laurence SC，Denslow ND，et al. Quercetin，a natural product supplement，impairs mitochondrial bioenergetics and locomotor behavior in larval zebrafish（Danio rerio） [J]. Toxicol Appl Pharmacol，2017，327：30-38.
[9] Qiu L，Luo Y，Chen X. Quercetin attenuates mitochondrial dysfunction and biogenesis via upregulated AMPK/SIRT1 signaling pathway in OA rats [J]. Biomed Pharmacother，2018， 103：1585-1591.
[10] Feng K，Chen Z，Pengcheng L，et al. Quercetin attenuates oxidative stress-induced apoptosis via SIRT1/AMPK-mediated inhibition of ER stress in rat chondrocytes and prevents the progression of osteoarthritis in a rat model [J]. J Cell Physiol 2019，234（10）：18192-18205.
[11] Li YC，Yeh CH，Yang ML，et al. Luteolin Suppresses Inflammatory Mediator Expression by Blocking the Akt/NF κB Pathway in Acute Lung Injury Induced by Lipop- olysaccharide in Mice [J]. Evid Based Complement Alternat Med，2012，2012：383608.
[12] Lee SA，Park BR，Moon SM，et al. Chondroprotective Effect of Cynaroside in IL-1beta-Induced Primary Rat Chondrocytes and Organ Explants via NF-κB and MAPK Signaling Inhibition [J]. Oxid Med Cell Longev，2020，2020： 9358080.
[13] Fei J，Liang B，Jiang C，et al. Luteolin inhibits IL-1β-induced inflammation in rat chondrocytes and T attenuates osteoarthritis progression in a rat model [J]. Biomed Pharmacother，2019，109：1586-1592.
[14] Zhou Z，Zhang L，Liu Y，et al. Luteolin Protects Chondrocytes from H2O2-Induced Oxidative Injury and Attenuates Osteoarthritis Progression by Activating AMPK-Nrf2 Signaling [J]. Oxid Med Cell Longev，2022，2022：5635797.
[15] Filomeni G，Desideri E，Cardaci S，et al. Carcinoma cells activate AMP-activated protein kinase-dependent autophagy as survival response to kaempferol-mediated energetic impairment [J]. Autophagy，2010，6（2）：202-216.
[16] Zhuang Z，Ye G，Huang B. Kaempferol Alleviates the Interleukin-1β-Induced Inflammation in Rat Osteoarthritis Chondrocytes via Suppression of NF-κB [J]. Med Sci Monit 2017，23：3925-3931.
[17] Tie F，Ding J，Hu N，et al. Kaempferol and Kaempferide Attenuate Oleic Acid-Induced Lipid Accumulation and Oxidative Stress in HepG2 Cells [J]. Int J Mol Sci，2021，22（16）：8847.
[18] Huang X，Pan Q，Mao Z，et al. Kaempferol inhibits interleukin1beta stimulated matrix metalloproteinases by suppressing the MAPK associated ERK and P38 signaling pathways [J]. Mol Med Rep，2018，18（3）：2697-2704.
[19] Jiang R，Hao P，Yu G，et al. Kaempferol protects chondrogenic ATDC5 cells against inflammatory injury triggered by lipopolysaccharide through down-regulating miR-146a [J]. Int Immunopharmacol，2019，69：373-381.
[20] Khan NM，Haseeb A，Ansari MY，et al. A wogonin-rich- fraction of Scutellaria baicalensis root extract exerts chondroprotective effects by suppressing IL-1β-induced activation of AP-1 in human OA chondrocytes [J]. Sci Rep 2017，7：43789.
[21] 沈晓庆，王晶，李婷君.汉黄芩素对胶原诱导性关节炎大鼠的治疗作用及对NLRP3炎症小体的影响[J].中医药导报，2021，27（8）：26-30.
[22] Khan NM，Ahmad I，Ansari MY，et al. Wogonin a natural flavonoid intercalates with genomic DNA and exhibits protective effects in IL-1β stimulated osteoarthritis chondrocytes [J]. Chem Biol Interact，2017，274：13-23.
[23] Yang H，Wang Z，Wang L，et al. Scutellarin ameliorates osteoarthritis by protecting chondrocytes and subchondral bone microstructure by inactivating NF-κB/MAPK signal transduction [J]. Biomed Pharmacother，2022，155：113781.
[24] 吴鹏，时孝晴，廖太阳，等.羌活-独活治疗膝骨关节炎的作用机制[J].中国骨质疏松杂志，2022，28（2）：262-268.
[25] 郭秋岩，李玮婕，王超，等.乌头汤缓解神经病理性疼痛的炎症网络调控机制研究[J].药学学报，2019，54（6）：1054-1061.
[26] Takada K，Hirose J，Senba K，et al. Enhanced apoptotic and reduced protective response in chondrocytes following endoplasmic reticulum stress in osteoarthritic cartilage [J]. Int J Exp Pathol，2011，92（4）：232-242.
[27] Hunter DJ，Bierma-Zeinstra S. Osteoarthritis [J]. Lancet，2019， 393（10182）：1745-1759.
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