Baicalin promotes extracellular matrix synthesis in chondrocytes via the activation of hypoxia‑inducible factor‑1&alpha;

Wang,Pengzhen; Zhu,Pingping; Liu,Ruijia; Meng,Qingqi; Li,Siming

doi:10.3892/etm.2020.9356

Baicalin promotes extracellular matrix synthesis in chondrocytes via the activation of hypoxia‑inducible factor‑1α

Authors:
- Pengzhen Wang
- Pingping Zhu
- Ruijia Liu
- Qingqi Meng
- Siming Li
View Affiliations

Affiliations: Guangzhou Institute of Traumatic Surgery, Guangzhou Red Cross Hospital, Medical College, Jinan University, Guangzhou, Guangdong 510220, P.R. China, Department of Internal Neurology, Guangzhou Red Cross Hospital, Medical College, Jinan University, Guangzhou, Guangdong 510220, P.R. China, Department of Orthopedics, Guangzhou Red Cross Hospital, Medical College, Jinan University, Guangzhou, Guangdong 510220, P.R. China
Published online on: October 15, 2020 https://doi.org/10.3892/etm.2020.9356
Article Number: 226

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Chinese herbal extracts are being used increasingly to treat osteoarthritis (OA) in recent years. Baicalin (BA) is an active component of Scutellaria baicalensis Georgi extracts and protects chondrocytes against damage. The aim of the present study was to examine the mechanism of action of BA on chondrocytes from mouse articular cartilage. In total, 44 µM BA and 10 µM hypoxia‑inducible‑factor‑1α (HIF‑1α) inhibitor BAY‑87‑2243 were screened by the [3‑(4,5‑dimethylthiazol‑2‑yl)‑5‑(3‑carboxymethoxyphenyl)‑2‑(4‑sulfophenyl)‑2H‑tetrazolium] method. Alcian blue and Safran O staining were used to investigate the synthesis of extracellular matrix (ECM) in chondrocytes treated with BA. The expression of HIF‑1α and chondrogenic marker genes including SOX9, AGG and Col2α was detected by western blotting or reverse‑transcription quantitative (RT‑qPCR), the expression of PHD1,2,3 and catabolic genes including ADAMTS5, MMP9 and MMP13 were detected by RT‑qPCR. To investigate the effect of BA on the ECM synthesis of chondrocytes, 44 µM BA and 10 µM BAY were chosen for further experimentation. It was confirmed that BA at a concentration of 44 µM could significantly promote the secretion of ECM. The expressions of genes including HIF‑1α, SOX9, collagen type 2 (Col2α) and aggrecan (AGG) were elevated following BA pretreatment and decreased by subsequent BAY‑87‑2243 stimulation for 24 h. Compared with untreated chondrocytes, the expressions of genes including ADAMTS5, MMP9, MMP13, PHD1, PHD2 and PHD3 in chondrocytes treated by BA were downregulated, however, BAY‑87‑2243 reversed the effect of BA on the genes including ADAMTS5, MMP9, MMP13, PHD1, PHD2 and PHD3 in chondrocytes. The findings of the present study suggest that BA may promote ECM synthesis and marker gene expression in chondrocytes by activating HIF‑1α. Therefore, BA may represent a novel clinical drug for OA.

View Figures

View References

1	Thomas AC, Hubbard-Turner T, Wikstrom EA and Palmieri-Smith RM: Epidemiology of posttraumatic osteoarthritis. J Athl Train. 52:491–496. 2017.PubMed/NCBI View Article : Google Scholar
2	Centers for Disease Control and Prevention (CDC): Public health and aging: projected prevalence of self-reported arthritis or chronic joint symptoms among persons aged >65 years--United States, 2005-2030. MMWR Morb Mortal Wkly Rep. 52(21):489-491, 2003.
3	Zhou X, Tenaglio S, Esworthy T, Hann SY, Cui H, Webster TJ, Fenniri H and Zhang LG: Three-Dimensional printing biologically inspired DNA-based gradient scaffolds for cartilage tissue regeneration. ACS Appl Mater Interfaces. 12:33219–33228. 2020.PubMed/NCBI View Article : Google Scholar
4	Frenkel SR, Clancy RM, Ricci JL, Di Cesare PE, Rediske JJ and Abramson SB: Effects of nitric oxide on chondrocyte migration E, adhesion, and cytoskeletal assembly. Arthritis Rheum. 39:1905–1912. 2015.PubMed/NCBI View Article : Google Scholar
5	Heinemeier KM, Schjerling P, Heinemeier J, Møller MB, Krogsgaard MR, Grum-Schwensen T, Petersen MM and Kjaer M: Radiocarbon dating reveals minimal collagen turnover in both healthy and osteoarthritic human cartilage. Sci Transl Med. 8(346ra90)2016.PubMed/NCBI View Article : Google Scholar
6	Oda K and Minata M: Drug free remission after steroid-dependent disappearance of lymphoproliferative disorder in rheumatoid arthritis patient treated with TNF-alpha blockade: Case study. Springerplus. 4(41)2015.PubMed/NCBI View Article : Google Scholar
7	Yammani RR, Carlson CS, Bresnick AR and Loeser RF: Increase in production of matrix metalloproteinase 13 by human articular chondrocytes due to stimulation with S100A4: Role of the receptor for advanced glycation end products. Arthritis Rheum. 54:2901–2911. 2006.PubMed/NCBI View Article : Google Scholar
8	Thomas CM, Fuller CJ, Whittles CE and Sharif M: Chondrocyte death by apoptosis is associated with cartilage matrix degradation. Osteoarthritis Cartilage. 15:27–34. 2007.PubMed/NCBI View Article : Google Scholar
9	Goldring MB, Fukuo K, Birkhead JR, Dudek E and Sandell LJ: Transcriptional suppression by interleukin-1 and interferon-gamma of type II collagen gene expression in human chondrocytes. J Cell Biochem. 54:85–99. 1994.PubMed/NCBI View Article : Google Scholar
10	Zheng W, Tao Z, Chen C, Zhang C, Zhang H, Ying X and Chen H: Plumbagin prevents IL-1β-induced inflammatory response in human osteoarthritis chondrocytes and prevents the progression of osteoarthritis in mice. Inflammation. 40:849–860. 2017.PubMed/NCBI View Article : Google Scholar
11	Xue M, McKelvey K, Shen K, Minhas N, March L, Park SY and Jackson CJ: Endogenous MMP-9 and not MMP-2 promotes rheumatoid synovial fibroblast survival, inflammation and cartilage degradation. Rheumatology (Oxford). 53:2270–2279. 2014.PubMed/NCBI View Article : Google Scholar
12	Alvarez J, Horton J, Sohn P and Serra R: The perichondrium plays an important role in mediating the effects of TGF-beta1 on endochondral bone formation. Dev Dyn. 221:311–321. 2001.PubMed/NCBI View Article : Google Scholar
13	Kim BS, Kang KS and Kang SK: Soluble factors from ASCs effectively direct control of chondrogenic fate. Cell Prolif. 43:249–261. 2010.PubMed/NCBI View Article : Google Scholar
14	Rodenas-Rochina J, Kelly DJ, Gómez Ribelles JL and Lebourg M: Influence of oxygen levels on chondrogenesis of porcine mesenchymal stem cells cultured in polycaprolactone scaffolds. J Biomed Mater Res A. 105:1684–1691. 2017.PubMed/NCBI View Article : Google Scholar
15	Zhang FJ, Luo W and Lei GH: Role of HIF-1α and HIF-2α in osteoarthritis. Joint Bone Spine. 82:144–147. 2015.PubMed/NCBI View Article : Google Scholar
16	Hu S, Zhang C, Ni L, Huang C, Chen D, Shi K, Jin H, Zhang K, Li Y, Xie L, et al: Stabilization of HIF-1α alleviates osteoarthritis via enhancing mitophagy. Cell Death Dis. 11(481)2020.PubMed/NCBI View Article : Google Scholar
17	Hirota K and Semenza GL: Regulation of hypoxia-inducible factor 1 by prolyl and asparaginyl hydroxylases. Biochem Biophys Res Commun. 338:610–616. 2005.PubMed/NCBI View Article : Google Scholar
18	Araldi E and Schipani E: Hypoxia, HIFs and bone development. Bone. 47:190–196. 2010.PubMed/NCBI View Article : Google Scholar
19	Denko NC: Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nat Rev Cancer. 8:705–713. 2008.PubMed/NCBI View Article : Google Scholar
20	Chen H, Xu Y, Wang J, Zhao W and Ruan H: Baicalin ameliorates isoproterenolinduced myocardial infarction through iNOS, inflammation and oxidative stress in rat. Int J Clin Exp Pathol. 8:10139–10147. 2015.PubMed/NCBI
21	Chen C, Zhang C, Cai L, Xie H, Hu W, Wang T, Lu D and Chen H: Baicalin suppresses IL-1β-induced expression of inflammatory cytokines via blocking NF-κB in human osteoarthritis chondrocytes and shows protective effect in mice osteoarthritis models. Int Immunopharmacol. 52:218–226. 2017.PubMed/NCBI View Article : Google Scholar
22	Zhu M, Ying J, Lin C, Wang Y, Huang K, Zhou Y and Teng H: Baicalin induces apoptotic death of human chondrosarcoma cells through mitochondrial dysfunction and downregulation of the PI3K/Akt/mTOR pathway. Planta Med. 85:360–369. 2019.PubMed/NCBI View Article : Google Scholar
23	Xing D, Gao H, Liu Z, Zhao Y and Gong M: Baicalin inhibits inflammatory responses to interleukin-1β stimulation in human chondrocytes. J Interferon Cytokine Res. 37:398–405. 2017.PubMed/NCBI View Article : Google Scholar
24	Idelevich A, Rais Y and Monsonego-Ornan E: Bone Gla protein increases HIF-1alpha-dependent glucose metabolism and induces cartilage and vascular calcification. Arterioscler Thromb Vasc Biol. 31:e55–e71. 2011.PubMed/NCBI View Article : Google Scholar
25	Pan Y, Chen D, Lu Q, Liu L, Li X and Li Z: Baicalin prevents the apoptosis of endplate chondrocytes by inhibiting the oxidative stress induced by H2O2. Mol Med Rep. 16:2985–2991. 2017.PubMed/NCBI View Article : Google Scholar
26	Griggio V, Vitale C, Todaro M, Riganti C, Kopecka J, Salvetti C, Bomben R, Bo MD, Magliulo D, Rossi D, et al: HIF-1α is over-expressed in leukemic cells from TP53-disrupted patients and is a promising therapeutic target in chronic lymphocytic leukemia. Haematologica. 105:1042–1054. 2020.PubMed/NCBI View Article : Google Scholar
27	Samarpita S, Doss HM, Ganesan R and Rasool M: Interleukin 17 under hypoxia mimetic condition augments osteoclast mediated bone erosion and expression of HIF-1α and MMP-9. Cell Immunol. 332:39–50. 2018.PubMed/NCBI View Article : Google Scholar
28	Gosset M, Berenbaum F, Thirion S and Jacques C: Primary culture and phenotyping of murine chondrocytes. Nat Protoc. 3:1253–1260. 2008.PubMed/NCBI View Article : Google Scholar
29	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
30	Chen C, Zhang C, Cai L, Xie H, Hu W, Wang T, Lu D and Chen H: Baicalin suppresses IL-1β-induced expression of inflammatory cytokines via blocking NF-κB in human osteoarthritis chondrocytes and shows protective effect in mice osteoarthritis models. Int Immunopharmacol. 52:218–226. 2017.PubMed/NCBI View Article : Google Scholar
31	He Y, Moqbel SAA, Xu L, Ran J, Ma C, Xu K, Bao J, Jiang L, Chen W, Xiong Y and Wu L: Costunolide inhibits matrix metalloproteinases expression and osteoarthritis via the NF-κB and Wnt/β-catenin signaling pathways. Mol Med Rep. 20:312–322. 2019.PubMed/NCBI View Article : Google Scholar
32	Huang X, Wu H, Wang L, Zheng L and Zhao J: Protective effects of baicalin on rabbit articular chondrocytes in vitro. Exp Ther Med. 13:1267–1274. 2017.PubMed/NCBI View Article : Google Scholar
33	Ellinghaus P, Heisler I, Unterschemmann K, Haerter M, Beck H, Greschat S, Ehrmann A, Summer H, Flamme I, Oehme F, et al: BAY 87-2243, a highly potent and selective inhibitor of hypoxia-induced gene activation has antitumor activities by inhibition of mitochondrial complex I. Cancer Med. 2:611–624. 2013.PubMed/NCBI View Article : Google Scholar
34	Robins JC, Akeno N, Mukherjee A, Dalal RR, Aronow BJ, Koopman P and Clemens TL: Hypoxia induces chondrocyte-specific gene expression in mesenchymal cells in association with transcriptional activation of Sox9. Bone. 37:313–322. 2005.PubMed/NCBI View Article : Google Scholar
35	Lefebvre V and Dvir-Ginzberg M: SOX9 and the many facets of its regulation in the chondrocyte lineage. Connect Tissue Res. 58:2–14. 2017.PubMed/NCBI View Article : Google Scholar
36	Duval E, Baugé C, Andriamanalijaona R, Bénateau H, Leclercq S, Dutoit S, Poulain L, Galéra P and Boumédiene K: Molecular mechanism of hypoxia-induced chondrogenesis and its application in in vivo cartilage tissue engineering. Biomaterials. 33:6042–6051. 2012.PubMed/NCBI View Article : Google Scholar
37	Wang P, Zhang F, He Q, Wang J, Shiu HT, Shu Y, Tsang WP, Liang S, Zhao K and Wan C: Flavonoid compound icariin activates hypoxia inducible factor-1α in chondrocytes and promotes articular cartilage repair. PLoS One. 11(e0148372)2016.PubMed/NCBI View Article : Google Scholar
38	Park SS, Bae I and Lee YJ: Flavonoids-induced accumulation of hypoxia-inducible factor (HIF)-1alpha/2alpha is mediated through chelation of iron. J Cell Biochem. 103:1989–1998. 2008.PubMed/NCBI View Article : Google Scholar
39	Leopoldini M, Russo N, Chiodo S and Toscano M: Iron chelation by the powerful antioxidant flavonoid quercetin. J Agric Food Chem. 54:6343–6351. 2006.PubMed/NCBI View Article : Google Scholar
40	Davis CK, Jain SA, Bae ON, Majid A and Rajanikant GK: Hypoxia mimetic agents for ischemic stroke. Front Cell Dev Biol. 6(175)2019.PubMed/NCBI View Article : Google Scholar
41	Terkhorn SP, Bohensky J, Shapiro IM, Koyama E and Srinivas V: Expression of HIF prolyl hydroxylase isozymes in growth plate chondrocytes: Relationship between maturation and apoptotic sensitivity. J Cell Physiol. 210:257–265. 2007.PubMed/NCBI View Article : Google Scholar
42	Wang W, Xu B, Xuan H, Ge Y, Wang Y, Wang L, Huang J, Fu W, Michie SA and Dalman RL: Hypoxia-inducible factor 1 in clinical and experimental aortic aneurysm disease. J Vasc Surg. 68:1538–1550.e2. 2018.PubMed/NCBI View Article : Google Scholar
43	Watts ER and Walmsley SR: Inflammation and hypoxia: HIF and PHD isoform selectivity. Trends Mol Med. 25:33–46. 2019.PubMed/NCBI View Article : Google Scholar