{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T11:58:54Z","timestamp":1788350334597,"version":"build-2803163510"},"reference-count":44,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2024,4,30]],"date-time":"2024-04-30T00:00:00Z","timestamp":1714435200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Science and Technology","award":["111-2314-B-016-028-MY3"],"award-info":[{"award-number":["111-2314-B-016-028-MY3"]}]},{"name":"Ministry of Science and Technology","award":["PTH111060"],"award-info":[{"award-number":["PTH111060"]}]},{"name":"Ministry of Science and Technology","award":["PTH111061"],"award-info":[{"award-number":["PTH111061"]}]},{"name":"Ministry of Science and Technology","award":["PTH112058"],"award-info":[{"award-number":["PTH112058"]}]},{"name":"Ministry of Science and Technology","award":["A1111035"],"award-info":[{"award-number":["A1111035"]}]},{"name":"Taoyuan General Hospital, Ministry of Health and Welfare","award":["111-2314-B-016-028-MY3"],"award-info":[{"award-number":["111-2314-B-016-028-MY3"]}]},{"name":"Taoyuan General Hospital, Ministry of Health and Welfare","award":["PTH111060"],"award-info":[{"award-number":["PTH111060"]}]},{"name":"Taoyuan General Hospital, Ministry of Health and Welfare","award":["PTH111061"],"award-info":[{"award-number":["PTH111061"]}]},{"name":"Taoyuan General Hospital, Ministry of Health and Welfare","award":["PTH112058"],"award-info":[{"award-number":["PTH112058"]}]},{"name":"Taoyuan General Hospital, Ministry of Health and Welfare","award":["A1111035"],"award-info":[{"award-number":["A1111035"]}]},{"name":"Teh-Tzer Study Group for Human Medical Research Foundation","award":["111-2314-B-016-028-MY3"],"award-info":[{"award-number":["111-2314-B-016-028-MY3"]}]},{"name":"Teh-Tzer Study Group for Human Medical Research Foundation","award":["PTH111060"],"award-info":[{"award-number":["PTH111060"]}]},{"name":"Teh-Tzer Study Group for Human Medical Research Foundation","award":["PTH111061"],"award-info":[{"award-number":["PTH111061"]}]},{"name":"Teh-Tzer Study Group for Human Medical Research Foundation","award":["PTH112058"],"award-info":[{"award-number":["PTH112058"]}]},{"name":"Teh-Tzer Study Group for Human Medical Research Foundation","award":["A1111035"],"award-info":[{"award-number":["A1111035"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Osteoarthritis is more prevalent than any other form of arthritis and is characterized by the progressive mechanical deterioration of joints. Glucosamine, an amino monosaccharide, has been used for over fifty years as a dietary supplement to alleviate osteoarthritis-related discomfort. Silibinin, extracted from milk thistle, modifies the degree of glycosylation of target proteins, making it an essential component in the treatment of various diseases. In this study, we aimed to investigate the functional roles of glucosamine and silibinin in cartilage homeostasis using the TC28a2 cell line. Western blots showed that glucosamine suppressed the N-glycosylation of the gp130, EGFR, and N-cadherin proteins. Furthermore, both glucosamine and silibinin differentially decreased and increased target proteins such as gp130, Snail, and KLF4 in TC28a2 cells. We observed that both compounds dose-dependently induced the proliferation of TC28a2 cells. Our MitoSOX and DCFH-DA dye data showed that 1 \u00b5M glucosamine suppressed mitochondrial reactive oxygen species (ROS) generation and induced cytosol ROS generation, whereas silibinin induced both mitochondrial and cytosol ROS generation in TC28a2 cells. Our JC-1 data showed that glucosamine increased red aggregates, resulting in an increase in the red\/green fluorescence intensity ratio, while all the tested silibinin concentrations increased the green monomers, resulting in decreases in the red\/green ratio. We observed increasing subG1 and S populations and decreasing G1 and G2\/M populations with increasing amounts of glucosamine, while increasing amounts of silibinin led to increases in subG1, S, and G2\/M populations and decreases in G1 populations in TC28a2 cells. MTT data showed that both glucosamine and silibinin induced cytotoxicity in TC28a2 cells in a dose-dependent manner. Regarding endoplasmic reticulum stress, both compounds induced the expression of CHOP and increased the level of p-eIF2\u03b1\/eIF2\u03b1. With respect to O-GlcNAcylation status, glucosamine and silibinin both reduced the levels of O-GlcNAc transferase and hypoxia-inducible factor 1 alpha. Furthermore, we examined proteins and mRNAs related to these processes. In summary, our findings demonstrated that these compounds differentially modulated cellular proliferation, mitochondrial and cytosol ROS generation, the mitochondrial membrane potential, the cell cycle profile, and autophagy. Therefore, we conclude that glucosamine and silibinin not only mediate glycosylation modifications but also regulate cellular processes in human chondrocytes.<\/jats:p>","DOI":"10.3390\/ijms25094905","type":"journal-article","created":{"date-parts":[[2024,5,1]],"date-time":"2024-05-01T03:30:49Z","timestamp":1714534249000},"page":"4905","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Glucosamine and Silibinin Alter Cartilage Homeostasis through Glycosylation and Cellular Stresses in Human Chondrocyte Cells"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3797-4514","authenticated-orcid":false,"given":"Yu-Pao","family":"Hsu","sequence":"first","affiliation":[{"name":"Department of Orthopedics, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan City 330, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tsung-Hsi","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan City 330, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shu-Ting","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, National Defense Medical Center, Taipei City 114, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9305-921X","authenticated-orcid":false,"given":"Shih-Ming","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, National Defense Medical Center, Taipei City 114, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2683-3815","authenticated-orcid":false,"given":"Yi-Chou","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan City 330, Taiwan"},{"name":"Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei City 114, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2563-8679","authenticated-orcid":false,"given":"Chia-Chun","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, Tri-Service General Hospital, National Defense Medical Center, Taipei City 114, Taiwan"},{"name":"Institute of Preventive Medicine, National Defense Medical Center, New Taipei City 237, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,4,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.joca.2021.04.020","article-title":"Epidemiology of osteoarthritis","volume":"30","author":"Allen","year":"2022","journal-title":"Osteoarthr. 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