缺氧下间充质干细胞分泌的外泌体对软骨细胞损伤的影响

doi:10.3969/j.issn.1006-5725.2025.22.009

Abstract

Abstract:

Objective To investigate the effects of exosomes derived from hypoxia-treated mesenchymal stem cells on chondrocyte injury. Methods After mesenchymal stem cells were subjected to hypoxic treatment， the secreted exosomes were collected and co-cultured with IL-1β-stimulated chondrocytes. Cell viability was assessed using the CCK-8 assay， while apoptosis was evaluated by flow cytometry and the measurement of Caspase-3 and PARP activities. Intracellular levels of ROS， Fe²⁺， and MDA were quantified using commercial assay kits. The expression of GPX4， SLC7A11， and ACSL4 was analyzed at both mRNA and protein levels via qRT-PCR and Western blot， respectively. Additionally， the secretion of COL2A1， MMP13， ADAMTS5， TNF-α， IL-6， and PGE2 was determined by ELISA. Results Chondrocyte viability was significantly enhanced following the uptake of exosomes derived from hypoxia-treated mesenchymal stem cells （H-Exo）（P < 0.05）. IL-1β treatment reduced chondrocyte viability， increased Caspase-3 and PARP activities， and promoted apoptosis （P < 0.05）； however， H-Exo effectively reversed IL-1β-induced apoptotic effects. Furthermore， IL-1β markedly down-regulated the expression of GPX4 and SLC7A11， up-regulated ACSL4 expression， and elevated intracellular levels of ROS， Fe²⁺， and MDA （P < 0.05）， indicating the induction of ferroptosis. Both the ferroptosis inhibitor and H-Exo significantly attenuated IL-1β-triggered ferroptosis， and H-Exo counteracted the detrimental effects of IL-1β as well as those induced by a ferroptosis inducer. Additionally， IL-1β suppressed the expression of the chondrogenic marker COL2A1， up-regulated the catabolic enzymes MMP13 and ADAMTS5， and enhanced the secretion of pro-inflammatory cytokines TNF-α， IL-6， and PGE2 （P < 0.05）. Notably， H-Exo alleviated IL-1β-mediated inflammation and restored the balance between chondrogenic anabolism and catabolism. Conclusions Exosomes secreted by mesenchymal stem cells under hypoxic conditions can effectively inhibit chondrocyte apoptosis and ferroptosis， thereby alleviating cellular injury. These findings suggest that such exosomes exert a protective effect on chondrocytes and hold promise as a novel therapeutic strategy for cartilage repair.

Key words: osteoarthritis, cartilage defect, chondrocyte, exosome, ferroptosis

CLC Number:

R684.3

Shaochu CHEN,Ming GONG,Wang ZHANG,Jiawen WU,Guangxin HUANG,Yadong ZHANG. Effects of exosomes secreted from mesenchymal stem cells on chondrocyte injury under hypoxia[J]. The Journal of Practical Medicine, 2025, 41(22): 3529-3536.

Figures/Tables 5

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References 23

[1]	洪坤豪，吴淮，刘文刚，等. 血清五聚素3和C1q/TNF相关蛋白3对膝关节骨性关节炎患者合并骨质疏松的诊断价值［J］. 实用医学杂志，2022，38（16）：2046-2050.
[2]	贾祥，徐田杰，樊佳欣，等. 二甲双胍通过激活SIRT1/p53信号通路对骨关节炎大鼠关节软骨发挥保护作用［J］. 实用医学杂志，2024，40（23）：3306-3316.
[3]	ZHANG W， LUO M， XING Y， et al. M2 Macrophage-Derived Extracellular Vehicles-Loaded Hyaluronic Acid-Alginate Hydrogel for Treatment of Osteoarthritis［J］. Orthop Surg， 2025，17（6）：1867-1881. doi:10.1111/os.70059 doi: 10.1111/os.70059
[4]	MAMACHAN M， SHARUN K， BANU S A， et al. Mesenchymal stem cells for cartilage regeneration： Insights into molecular mechanism and therapeutic strategies［J］. Tissue Cell， 2024， 88：102380. doi:10.1016/j.tice.2024.102380 doi: 10.1016/j.tice.2024.102380
[5]	TAN F， LI X， WANG Z， et al. Clinical applications of stem cell-derived exosomes［J］. Signal Transduct Target Ther， 2024， 9（1）：17. doi:10.1038/s41392-023-01704-0 doi: 10.1038/s41392-023-01704-0
[6]	YU H， HUANG Y， YANG L. Research progress in the use of mesenchymal stem cells and their derived exosomes in the treatment of osteoarthritis［J］. Ageing Res Rev， 2022， 80：101684. doi:10.1016/j.arr.2022.101684 doi: 10.1016/j.arr.2022.101684
[7]	HAO W， ZHU R， ZHANG H， et al. NS8593 inhibits chondrocyte ferroptosis and alleviates cartilage injury in rat adjuvant arthritis through TRPM7/HO-1 pathway［J］. Int J Biochem Cell Biol， 2024， 174：106618. doi:10.1016/j.biocel.2024.106618 doi: 10.1016/j.biocel.2024.106618
[8]	XU W， ZHANG B， XI C， et al. Ferroptosis Plays a Role in Human Chondrocyte of Osteoarthritis Induced by IL-1β In Vitro［J］. Cartilage， 2023， 14（4）：455-466. doi:10.1177/19476035221142011 doi: 10.1177/19476035221142011
[9]	VINCENT T L， ALLISTON T， KAPOOR M， et al. Osteoarthritis Pathophysiology： Therapeutic Target Discovery may Require a Multifaceted Approach［J］. Clin Geriatr Med， 2022， 38（2）：193-219. doi:10.1016/j.cger.2021.11.015 doi: 10.1016/j.cger.2021.11.015
[10]	TUNCAY DURUÖZ M， ÖZ N， GÜRSOY D E， et al. Clinical aspects and outcomes in osteoarthritis［J］. Best Pract Res Clin Rheumatol， 2023， 37（2）：101855. doi:10.1016/j.berh.2023.101855 doi: 10.1016/j.berh.2023.101855
[11]	XIAO S Q， CHENG M， WANG L， et al. The role of apoptosis in the pathogenesis of osteoarthritis［J］. Int Orthop， 2023， 47（8）：1895-1919. doi:10.1007/s00264-023-05847-1 doi: 10.1007/s00264-023-05847-1
[12]	AL-HETTY H R A K， ABDULAMEER S J， ALGHAZALI M W， et al. The Role of Ferroptosis in the Pathogenesis of Osteoarthritis［J］. J Membr Biol， 2023， 256（3）：223-228. doi:10.1007/s00232-023-00282-0 doi: 10.1007/s00232-023-00282-0
[13]	LIU Y， ZHANG Z， FANG Y， et al. Ferroptosis in Osteoarthritis： Current Understanding［J］. J Inflamm Res， 2024， 17：8471-8486. doi:10.2147/jir.s493001 doi: 10.2147/jir.s493001
[14]	PAN X， KONG X， FENG Z， et al. 4-Octyl itaconate protects chondrocytes against IL-1β-induced oxidative stress and ferroptosis by inhibiting GPX4 methylation in osteoarthritis［J］. Int Immunopharmacol， 2024， 137：112531. doi:10.1016/j.intimp.2024.112531 doi: 10.1016/j.intimp.2024.112531
[15]	LIU J， ZHOU H， CHEN J， et al. Baicalin inhibits IL-1β-induced ferroptosis in human osteoarthritis chondrocytes by activating Nrf-2 signaling pathway［J］. J Orthop Surg Res， 2024， 19（1）：23. doi:10.1186/s13018-023-04483-0 doi: 10.1186/s13018-023-04483-0
[16]	TIAN R， SU S， YU Y， et al. Revolutionizing osteoarthritis treatment： How mesenchymal stem cells hold the key［J］. Biomed Pharmacother， 2024， 173：116458. doi:10.1016/j.biopha.2024.116458 doi: 10.1016/j.biopha.2024.116458
[17]	HADE M D， SUIRE C N， SUO Z. Mesenchymal Stem Cell-Derived Exosomes： Applications in Regenerative Medicine［J］. Cells， 2021， 10（8）：1959. doi:10.3390/cells10081959 doi: 10.3390/cells10081959
[18]	SHEN K， DUAN A， CHENG J， et al. Exosomes derived from hypoxia preconditioned mesenchymal stem cells laden in a silk hydrogel promote cartilage regeneration via the miR-205-5p/PTEN/AKT pathway［J］. Acta Biomater， 2022， 143：173-188. doi:10.1016/j.actbio.2022.09.005 doi: 10.1016/j.actbio.2022.09.005
[19]	ZHAO J， SUN Y， SHENG X， et al. Hypoxia-treated adipose mesenchymal stem cell-derived exosomes attenuate lumbar facet joint osteoarthritis［J］. Mol Med， 2023， 29（1）：120. doi:10.1186/s10020-023-00709-3 doi: 10.1186/s10020-023-00709-3
[20]	XU W， ZHANG B， XI C， et al. Ferroptosis Plays a Role in Human Chondrocyte of Osteoarthritis Induced by IL-1β In Vitro［J］. Cartilage， 2023， 14（4）：455-466. doi:10.1177/19476035221142011 doi: 10.1177/19476035221142011
[21]	PENG S， SUN C， LAI C， et al. Exosomes derived from mesenchymal stem cells rescue cartilage injury in osteoarthritis through Ferroptosis by GOT1/CCR2 expression［J］. Int Immunopharmacol， 2023， 122：110566. doi:10.1016/j.intimp.2023.110566 doi: 10.1016/j.intimp.2023.110566
[22]	LIU Y， WANG Y， YANG M， et al. Exosomes from hypoxic pretreated ADSCs attenuate ultraviolet light-induced skin injury via GLRX5 delivery and ferroptosis inhibition［J］. Photochem Photobiol Sci， 2024， 23（1）：55-63. doi:10.1007/s43630-023-00498-y doi: 10.1007/s43630-023-00498-y
[23]	SHAO M， YE S， CHEN Y， et al. Exosomes from hypoxic ADSCs ameliorate neuronal damage post spinal cord injury through circ-Wdfy3 delivery and inhibition of ferroptosis［J］. Neurochem Int， 2024， 177：105759. doi:10.1016/j.neuint.2024.105759 doi: 10.1016/j.neuint.2024.105759

基因名称	引物序列
GPX4	F: 5′-GAGGCAAGACCGAAGTAAACTAC-3′
	R: 5′-CCGAACTGGTTACACGGGAA-3′
SLC7A11	F: 5′-TGCTGGGCTGATTTTATCTTCG-3′
	R: 5′-GAAAGGGCAACCATGAAGAGG-3′
ACSL4	F: 5′-TCTGCTTCTGCTGCCCAATT-3′
	R: 5′-CGCCTTCTTGCCAGTCTTTT-3′
COL2A1	F: 5′-TGCATGAGGGCGCGGTA-3′
	R: 5′-GGTCCTGGTTGCCGGACAT-3′
MMP13	F: 5′-ATGACTATGCGTGGCTGGAA-3′
	R: 5′-TGTCCCATTTGTGGTGTGGG-3′
ADAMTS5	F: 5′-TGGCTCACGAAATCGGACAT-3′
	R: 5′-TTGGACCAGGGCTTAGATGC-3′
GAPDH	F: 5'-CGCATCTTCTTTTGCGTCG-3'
	R: 5'-TTGAGGTCAATGAAGGGGTCA-3'

Effects of exosomes secreted from mesenchymal stem cells on chondrocyte injury under hypoxia

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