Glucosamine Protects Nucleus Pulposus Cells and Induces Autophagy via the mTOR-Dependent Pathway
Glucosamine Protects Nucleus Pulposus Cells and Induces Autophagy via the mTOR-Dependent Pathway
复制标题
葡萄糖胺通过 mTOR 依赖性途径保护髓核细胞并诱导自噬
DOI：
10.1002/jor.22699
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发表时间：
2014-11-01
影响因子：
2.8
通讯作者：
Dong, Jian
中科院分区：
文献类型：
--
作者：
Jiang, LiBo;Jin, YongLong;Dong, Jian
虽然氨基葡萄糖已被认为是有效的治疗骨关节炎，其对椎间盘退变的影响仍不清楚。我们试图探索葡萄糖胺是否可以激活大鼠髓核（NP）细胞中的自噬，并保护用IL-1或过氧化氢（H2 O2）处理的细胞。通过检测LC 3、Beclin-1、m-TOR和p70 S6 K以及通过分析自噬体来检查细胞中的自噬。为了抑制自噬，使用3-甲基腺嘌呤（3-MA）。在用IL-1处理的细胞中，通过实时PCR和免疫荧光分析Adamts-4、Mmp-13、聚集蛋白聚糖和Col 2a 1的水平。TUNEL法检测细胞凋亡。SA-Gal染色显示H_2O_2诱导的细胞衰老。氨基葡萄糖可在24 h内剂量依赖性地激活自噬，并抑制m-TOR和p70 S6 K的磷酸化。自噬在IL-1或H2 O2处理的细胞增加葡糖胺。氨基葡萄糖可减轻IL-1诱导的NP细胞聚集蛋白聚糖的减少和凋亡，而3-MA可部分逆转这些作用。氨基葡萄糖可降低H_2O_2诱导的SA-Gal阳性细胞百分率，并伴有p70 S6 K磷酸化水平的下降。氨基葡萄糖通过抑制m-TOR通路保护NP细胞并上调自噬，这可能是椎间盘退变的潜在治疗剂。(c)2014骨科研究学会。出版社：Wiley Periodicals，Inc. J Orthop Res 32：1532-1542，2014.
Although glucosamine has been suggested to be effective in the treatment of osteoarthritis, its effect on disc degeneration remains unclear. We sought to explore whether glucosamine can activate autophagy in rat nucleus pulposus (NP) cells and protect cells treated with IL-1 or hydrogen peroxide (H2O2). Autophagy in cells was examined by detecting for LC3, Beclin-1, m-TOR, and p70S6K, as well as by analyzing autophagosomes. To inhibit autophagy, 3-methyladenine (3-MA) was used. In the cells treated with IL-1, the levels of Adamts-4, Mmp-13, aggrecan, and Col2a1 were analyzed by real-time PCR and immunofluorescence. Apoptosis was analyzed by TUNEL. Cell senescence under H2O2 was revealed by SA--Gal staining. Glucosamine could activate autophagy in a dose-dependent manner within 24h and inhibit the phosphorylation of m-TOR and p70S6K. Autophagy in IL-1 or H2O2-treated cells was increased by glucosamine. Glucosamine attenuated the decrease of aggrecan and prevented the apoptosis of the NP cells induced by IL-1, whereas 3-MA partly reversed these effects. The percentage of SA--Gal-positive cells induced by H2O2 treatment was decreased by glucosamine, accompanied by the decline of p70S6K phosphorylation. Glucosamine protects NP cells and up-regulates autophagy by inhibiting the m-TOR pathway, which might point a potential therapeutic agent for disc degeneration. (c) 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:1532-1542, 2014.