PMID- 36940989 OWN - NLM STAT- MEDLINE DCOM- 20230322 LR - 20230323 IS - 1002-1892 (Print) IS - 1002-1892 (Linking) VI - 37 IP - 3 DP - 2023 Mar 15 TI - [Research of epigallocatechin gallate in delaying chondrocyte senescence]. PG - 308-315 LID - 10.7507/1002-1892.202210101 [doi] AB - OBJECTIVE: To investigate the effect of epigallocatechin gallate (EGCG) on chondrocyte senescence and its mechanism. METHODS: The chondrocytes were isolated from the articular cartilage of 4-week-old Sprague Dawley rats, and cultured with type Ⅱcollagenase and passaged. The cells were identified by toluidine blue staining, alcian blue staining, and immunocytochemical staining for type Ⅱ collagen. The second passage (P2) cells were divided into blank control group, 10 ng/mL IL-1beta group, and 6.25, 12.5, 25.0, 50.0, 100.0, and 200.0 mumol/L EGCG+10 ng/mL IL-1beta group. The chondrocyte activity was measured with cell counting kit 8 after 24 hours of corresponding culture, and the optimal drug concentration of EGCG was selected for the subsequent experiment. The P2 chondrocytes were further divided into blank control group (group A), 10 ng/mL IL-1beta group (group B), EGCG+10 ng/mL IL-1beta group (group C), and EGCG+10 ng/mL IL-1beta+5 mmol/L 3-methyladenine (3-MA) group (group D). After cultured, the degree of cell senescence was detected by beta-galactosidase staining, the autophagy by monodansylcadaverine method, and the expression levels of chondrocyte-related genes [type Ⅱ collagen, matrix metalloproteinase 3 (MMP-3), MMP-13] by real-time fluorescent quantitative PCR, the expression levels of chondrocyte-related proteins (Beclin-1, LC3, MMP-3, MMP-13, type Ⅱ collagen, P16, mTOR, AKT) by Western blot. RESULTS: The cultured cells were identified as chondrocytes. Compared with the blank control group, the cell activity of 10 ng/mL IL-1beta group significantly decreased ( P<0.05). Compared with the 10 ng/mL IL-1beta group, the cell activity of EGCG+10 ng/mL IL-1beta groups increased, and the 50.0, 100.0, and 200.0 mumol/L EGCG significantly promoted the activity of chondrocytes ( P<0.05). The 100.0 mumol/L EGCG was selected for subsequent experiments. Compared with group A, the cells in group B showed senescence changes. Compared with group B, the senescence rate of chondrocytes in group C decreased, autophagy increased, the relative expression of type Ⅱ collagen mRNA increased, and relative expressions of MMP-3 and MMP-13 mRNAs decreased; the relative expressions of Beclin-1, LC3, and type Ⅱ collagen proteins increased, but the relative expressions of P16, MMP-3, MMP-13, mTOR, and AKT proteins decreased; the above differences were significant ( P<0.05). Compared with group C, when 3-MA was added in group D, the senescence rate of chondrocytes increased, autophagy decreased, and the relative expressions of the target proteins and mRNAs showed an opposite trend ( P<0.05). CONCLUSION: EGCG regulates the autophagy of chondrocytes through the PI3K/AKT/mTOR signaling pathway and exerts anti-senescence effects. FAU - Luo, Chao AU - Luo C AD - Department of Orthopedics, the Second Affiliated Hospital of Nanchang University, Nanchang Jiangxi, 330006, P. R. China. AD - Department of Orthopedics, Nanchang County People's Hospital, Nanchang Jiangxi, 330200, P. R. China. FAU - Zhou, Weijun AU - Zhou W AD - Department of Orthopedics, the Second Affiliated Hospital of Nanchang University, Nanchang Jiangxi, 330006, P. R. China. FAU - Zhang, Huying AU - Zhang H AD - Department of Orthopedics, the Second Affiliated Hospital of Nanchang University, Nanchang Jiangxi, 330006, P. R. China. FAU - Zhu, Jinwei AU - Zhu J AD - Department of Orthopedics, the Second Affiliated Hospital of Nanchang University, Nanchang Jiangxi, 330006, P. R. China. FAU - Chen, Lu AU - Chen L AD - Department of Orthopedics, the Second Affiliated Hospital of Nanchang University, Nanchang Jiangxi, 330006, P. R. China. FAU - Gu, Yurong AU - Gu Y AD - Department of Orthopedics, the Second Affiliated Hospital of Nanchang University, Nanchang Jiangxi, 330006, P. R. China. LA - chi PT - English Abstract PT - Journal Article PL - China TA - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi JT - Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery JID - 9425194 RN - EC 3.4.24.17 (Matrix Metalloproteinase 3) RN - EC 3.4.24.- (Matrix Metalloproteinase 13) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - 0 (Collagen Type II) RN - BQM438CTEL (epigallocatechin gallate) RN - EC 2.7.1.- (Phosphatidylinositol 3-Kinases) RN - 0 (Beclin-1) RN - 0 (Interleukin-1beta) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - 0 (RNA, Messenger) SB - IM MH - Rats MH - Animals MH - Rats, Sprague-Dawley MH - *Matrix Metalloproteinase 3/genetics/metabolism/pharmacology MH - *Chondrocytes/metabolism MH - Matrix Metalloproteinase 13/genetics/metabolism MH - Proto-Oncogene Proteins c-akt/metabolism MH - Collagen Type II/genetics/metabolism MH - Phosphatidylinositol 3-Kinases/metabolism/pharmacology MH - Beclin-1/metabolism MH - Interleukin-1beta/metabolism MH - TOR Serine-Threonine Kinases/metabolism/pharmacology MH - RNA, Messenger MH - Cells, Cultured PMC - PMC10027523 OTO - NOTNLM OT - Epigallocatechin gallate OT - PI3K/AKT/mTOR pathway OT - autophagy OT - chondrocytes OT - senescence COIS- 利益冲突　在课题研究和文章撰写过程中不存在利益冲突 EDAT- 2023/03/21 06:00 MHDA- 2023/03/23 06:00 PMCR- 2023/03/01 CRDT- 2023/03/20 20:34 PHST- 2023/03/20 20:34 [entrez] PHST- 2023/03/21 06:00 [pubmed] PHST- 2023/03/23 06:00 [medline] PHST- 2023/03/01 00:00 [pmc-release] AID - zgxfcjwkzz-37-3-308 [pii] AID - 10.7507/1002-1892.202210101 [doi] PST - ppublish SO - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2023 Mar 15;37(3):308-315. doi: 10.7507/1002-1892.202210101.