Protective effect of astragaloside IV against inflammatory injury in chondrocytes

doi:10.12307/2023.120

Abstract

Abstract: BACKGROUND: Cartilage lesions are the key to the occurrence and development of osteoarthritis. Exploring the function of chondrocytes and developing drugs for chondrocyte protection and pro-secretion of type II collagen are important means to clinically cure osteoarthritis.
OBJECTIVE: To explore the protective effect of astragaloside IV (AS-IV) on interleukin-1β-induced inflammation of chondrocytes and to explore whether the Hippo-YAP pathway is involved in this effect and its potential mechanism.
METHODS: Chondrocytes from the tibia and femur of Sprague-Dawley rats were isolated and cultured. Chondrocytes in the logarithmic growth phase were divided into five groups: blank group in which the cells were routinely cultured, model group in which the cells were treated with interleukin-1β to induce inflammatory injury in chondrocytes, astragaloside IV group in which the cells were treated with interleukin-1β + astragaloside IV, NC-siRNA group in which the cells were transfected with NC-siRNA + interleukin-1β + astragaloside IV, YAP1-siRNA group in which the cells were transfected with YAP1-siRNA+interleukin-1β+astragaloside IV. Relevant measurements were performed after 48 hours of culture.
RESULTS AND CONCLUSION: The cell viability was significantly lower in the model group than the blank group (P < 0.05), significantly higher in the astragaloside IV, NC-siRNA, and YAP1-siRNA groups than the model group (P < 0.05), and significantly higher in the YAP1-siRNA group than the NC-siRNA group (P < 0.05). The apoptotic rate was significantly higher in the model group than the blank group (P < 0.05), significantly lower in the astragaloside IV, NC-siRNA, and YAP1-siRNA transfection groups than the model group (P < 0.05), and significantly lower in the YAP1-siRNA group than the NC-siRNA group (P < 0.05). RT-qRCR results showed that compared with the blank group, the mRNA expressions of metalloproteinase 1 and tissue inhibitor of metalloproteinase 1 were significantly higher in the model group (P < 0.05), while the mRNA expressions of YAP1 and TEAD1 were significantly lower in the model group (P < 0.05). The mRNA expression of YAP1 in the astragaloside IV group was significantly higher than that in the model group (P < 0.05). Compared with the NC-siRNA group, the mRNA expressions of metalloproteinase 1 and tissue inhibitor of metalloproteinase 1 were significantly higher in the YAP1-siRNA group (P < 0.05), while the mRNA expressions of YAP1 and TEAD1 were significantly lower in the YAP1-siRNA group (P < 0.05). Western blot results revealed that compared with the blank group, the protein expressions of metalloproteinase 1, tissue inhibitor of metalloproteinase 1, and YAP1 were significantly higher in the model group (P < 0.05), while the protein expressions of type II collagen, TEAD1, and p-YAP1 were significantly lower in the model group (P < 0.05). Compared with the NC-siRNA group, the protein expressions of metalloproteinase 1 and tissue inhibitor of metalloproteinase 1 were significantly higher in the YAP1-siRNA group (P < 0.05), while the protein expressions of type II collagen, TEAD1, and p-YAP1 were significantly lower in the YAP1-siRNA group (P < 0.05). To conclude, astragaloside IV has a protective effect on interleukin-1β-induced inflammation of chondrocytes, which may be related to the Hippo-YAP pathway.

Key words: osteoarthritis, chondrocyte, astragaloside IV, Hippo-YAP, interleukin-1β, type II collagen

CLC Number:

Liao Jianzhao, Yang Nan, Zhou Yi, Xu Hang, Xia Tian, Song Shilei, Zeng Qi, Chen Yueping. Protective effect of astragaloside IV against inflammatory injury in chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(14): 2158-2163.

Figures/Tables 6

References

[1] 中华医学会骨科学分会关节外科学组.骨关节炎诊疗指南[J].中华骨科杂志,2018,38(12):705-715.
[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. Ann Rheum Dis. 2020;79(6):819-828.
[3] 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.
[4] ZHANG JF, SONG LH, WEI JN, et al. Prevalence of and risk factors for the occurrence of symptomatic osteoarthritis in rural regions of Shanxi Province, China. Int J Rheum Dis. 2016;19(8):781-789.
[5] ABRAMOFF B, CALDERA FE. Osteoarthritis: Pathology, Diagnosis, and Treatment Options. Med Clin North Am. 2020;104(2):293-311.
[6] LUO Y, SINKEVICIUTE D, HE Y, et al. The minor collagens in articular cartilage. Protein Cell. 2017;8(8):560-572.
[7] HUNTER DJ, BIERMA-ZEINSTRA S. Osteoarthritis. Lancet. 2019; 393(10182):1745-1759.
[8] MOYA IM, HALDER G. Hippo-YAP/TAZ signalling in organ regeneration and regenerative medicine. Nat Rev Mol Cell Biol. 2019;20(4):211-226.
[9] WANG S, ZHOU L, LING L, et al. The Crosstalk Between Hippo-YAP Pathway and Innate Immunity. Front Immunol. 2020;11:323.
[10] IBAR C, IRVINE KD. Integration of Hippo-YAP Signaling with Metabolism. Dev Cell. 2020;54(2):256-267.
[11] CHUANG LSH, ITO Y. The Multiple Interactions of RUNX with the Hippo-YAP Pathway. Cells. 2021;10(11):2925.
[12] 袁永刚.Yes相关蛋白(YAP)通过细胞外基质力学调节非小细胞肺癌生长的研究[D].济南:山东大学,2015.
[13] DENG Y, LU J, LI W, et al. Reciprocal inhibition of YAP/TAZ and NF-κB regulates osteoarthritic cartilage degradation. Nat Commun. 2018;9(1): 4564.
[14] WANG F, QIAN H, KONG L, et al. Accelerated Bone Regeneration by Astragaloside IV through Stimulating the Coupling of Osteogenesis and Angiogenesis. Int J Biol Sci. 2021;17(7):1821-1836.
[15] JIANG C, ZHOU Z, LIN Y, et al. Astragaloside IV ameliorates steroid-induced osteonecrosis of the femoral head by repolarizing the phenotype of pro-inflammatory macrophages. Int Immunopharmacol. 2021;93:107345.
[16] LI M, WANG W, GENG L, et al. Inhibition of RANKL-induced osteoclastogenesis through the suppression of the ERK signaling pathway by astragaloside IV and attenuation of titanium-particle-induced osteolysis. Int J Mol Med. 2015;36(5):1335-1344.
[17] HAN J, SHEN X, ZHANG Y, et al. Astragaloside IV suppresses transforming growth factor-β1-induced epithelial-mesenchymal transition through inhibition of Wnt/β-catenin pathway in glioma U251 cells. Biosci Biotechnol Biochem. 2020;84(7):1345-1352.
[18] 赵永见,薛纯纯,王利波,等.黄芪甲苷对关节不稳诱导型小鼠膝骨关节炎的保护作用[J].中国中医基础医学杂志,2015,21(6):668-670.
[19] 余素姣,谭慧.黄芪甲苷通过炎症小体活化影响软骨细胞炎性因子的表达[J].中国组织工程研究,2019,23(11):1652-1656.
[20] LIU J, MENG Q, JING H, et al. Astragaloside IV protects against apoptosis in human degenerative chondrocytes through autophagy activation. Mol Med Rep. 2017;16(3):3269-3275.
[21] RAHMATI M, NALESSO G, MOBASHERI A, et al. Aging and osteoarthritis: Central role of the extracellular matrix. Ageing Res Rev. 2017;40:20-30.
[22] CHARLIER E, DEROYER C, CIREGIA F, et al. Chondrocyte dedifferentiation and osteoarthritis (OA). Biochem Pharmacol. 2019;165:49-65.
[23] YANG Y, LIN H, SHEN H, et al. Mesenchymal stem cell-derived extracellular matrix enhances chondrogenic phenotype of and cartilage formation by encapsulated chondrocytes in vitro and in vivo. Acta Biomater. 2018;69:71-82.
[24] 刘建红,孟庆刚,胡佳卉,等.黄芪甲苷对早期骨关节炎软骨细胞基质金属蛋白酶-1表达的影响[J].中国老年学杂志,2017,37(3):532-534.
[25] CUI N, HU M, KHALIL RA. Biochemical and Biological Attributes of Matrix Metalloproteinases. Prog Mol Biol Transl Sci. 2017;147:1-73.
[26] TOMASZEWSKA E, RUDYK H, ŚWIETLICKA I, et al. Trabecular Bone Parameters, TIMP-2, MMP-8, MMP-13, VEGF Expression and Immunolocalization in Bone and Cartilage in Newborn Offspring Prenatally Exposed to Fumonisins. Int J Mol Sci. 2021;22(22):12528.
[27] SAW S, AIKEN A, FANG H, et al. Metalloprotease inhibitor TIMP proteins control FGF-2 bioavailability and regulate skeletal growth. J Cell Biol. 2019;218(9):3134-3152.
[28] AHARONOV A, SHAKKED A, UMANSKY KB, et al. ERBB2 drives YAP activation and EMT-like processes during cardiac regeneration. Nat Cell Biol. 2020;22(11):1346-1356.
[29] WANG J, LIU S, HEALLEN T, et al. The Hippo pathway in the heart: pivotal roles in development, disease, and regeneration. Nat Rev Cardiol. 2018;15(11):672-684.
[30] SUN X, REN Z, CUN Y, et al. Hippo-YAP signaling controls lineage differentiation of mouse embryonic stem cells through modulating the formation of super-enhancers. Nucleic Acids Res. 2020;48(13):7182-7196.
[31] LI C, JIN Y, WEI S, et al. Hippo Signaling Controls NLR Family Pyrin Domain Containing 3 Activation and Governs Immunoregulation of Mesenchymal Stem Cells in Mouse Liver Injury. Hepatology. 2019;70(5): 1714-1731.
[32] ZHOU X, LI Y, WANG W, et al. Regulation of Hippo/YAP signaling and Esophageal Squamous Carcinoma progression by an E3 ubiquitin ligase PARK2. Theranostics. 2020;10(21):9443-9457.
[33] LIU H, DU S, LEI T, et al. Multifaceted regulation and functions of YAP/TAZ in tumors (Review). Oncol Rep. 2018;40(1):16-28.
[34] LUO H, YAO L, ZHANG Y, et al. Liquid chromatography-mass spectrometry-based quantitative proteomics analysis reveals chondroprotective effects of astragaloside IV in interleukin-1β-induced SW1353 chondrocyte-like cells. Biomed Pharmacother. 2017;91:796-802.
[35] WANG L, WANG S, SHI Y, et al. YAP and TAZ protect against white adipocyte cell death during obesity. Nat Commun. 2020;11(1):5455.