姜黄素对人椎间盘髓核细胞自噬和退变的影响

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摘要目的探讨姜黄素对人椎间盘髓核细胞自噬和退变的影响。方法人椎间盘髓核组织来源于2018年3～11月四川省遂宁市中心医院骨科收治的7例椎间盘突出患者，并进行体外髓核细胞原代培养，将其分为三组：对照组、白细胞介素-1β（IL-1β）组和白细胞介素-1β+姜黄素组。四甲基偶氮唑盐比色法（MTT）检测姜黄素对人髓核细胞毒性情况，RT-PCR和Western blot分别检测各组细胞内基质金属蛋白酶-3（MMP-3）、MMP-13、聚集蛋白聚糖（aggrecan）和胶原蛋白（Collagen）Ⅱ的表达水平，同时检测各组细胞内自噬蛋白微管相关蛋白3（LC3）和Beclin 1的蛋白表达情况；采用GFP-LC3腺病毒转染观察各组细胞内自噬水平。结果 MTT结果显示，当姜黄素浓度＜ 80 μmol/L时对髓核细胞是没有细胞毒性的，考虑到对髓核细胞无毒性，同时对髓核细胞能发挥最大的药物作用，选择姜黄素浓度为80 μmol/L进行后续的实验。姜黄素能显著增加髓核细胞内的自噬颗粒数，促进细胞自噬蛋白LC3的水平升高和P62蛋白水平下降（P ＜ 0.05）；同时，姜黄素显著提高髓核细胞aggrecan和Collagen Ⅱ的水平，降低MMP-3和MMP-13的表达水平（P ＜ 0.05）。结论姜黄素能减缓椎间盘的退变，其机制可能是通过促进细胞的自噬水平。

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	万俊锋阳波郑佳状冉茂波宋昭君

关键词 ：姜黄素, 自噬, 髓核细胞, 退变

Abstract：Objective To investigate the effect of Curcumin on autophagy and degeneration of human intervertebral disc nucleus pulposus. Methods The human disc nucleus pulposus tissues were obtained from 7 patients with disc herniation admitted to the Department of Orthopedics, Suining Central Hospital of Sichuan Province from March to November 2018. Primary culture of nucleus pulposus cells were performed in vitro and divided into three groups, control group, interleukin-1β (IL-1β) group and interleukin-1β + Curcumin group. The toxicity of Curcumin to human nucleus pulposus cells was detected by tetramethylazoles salt colorimetry (MTT) assay; RT-PCR and Western blot were used respectively to detect each cell matrix metalloproteinases-3 (MMP-3), MMP-13, gathered proteoglycans (aggrecan) and collagen (Collagen) Ⅱ expression level; meanwhile, protein expressions of autophagy protein microtubule-related protein 3 (LC3) and Beclin 1 in each group were detected. The autophagy level in each group was observed by transfection with GFP-LC3 adenovirus. Results MTT results showed no cytotoxicity to nucleus pulposus cells when curcumin concentration was less than 80 μmol/L. Considering that it has no toxic effect on nucleus pulposus cells and can exert maximum drug effect on nucleus pulposus cells. Curcumin concentration of 80 μmol/L was selected for subsequent experiments. Curcumin can significantly increase the number of autophagy particles in nucleus pulposus cells, promote the increase of autophagy protein LC3 level and decrease of P62 protein level (P ＜ 0.05). Curcumin significantly improve aggrecan nucleus pulposus cells and Collagen Ⅱ level, reduce the MMP-3 and the expression of MMP-13 levels (P ＜ 0.05). Conclusion Curcumin can slow disc degeneration, possibly by promoting autophagy in cells.

Key words： Curcumin Autophagy Nucleus pulposus cells Degeneration

基金资助:四川省卫生和计划生育委员会科研课题（16PJ5 23）。

作者简介: 万俊锋（1990.1-），男，硕士；研究方向：椎间盘退变。

引用本文:

万俊锋阳波郑佳状冉茂波宋昭君. 姜黄素对人椎间盘髓核细胞自噬和退变的影响[J]. 中国医药导报, 2019, 16(25): 11-15.
WAN Junfeng YANG Bo ZHENG Jiazhuang RAN Maobo SONG Zhaojun. Effects of curcumin on autophagy and degeneration of human intervertebral disc nucleus pulposus. 中国医药导报, 2019, 16(25): 11-15.

链接本文:

http://www.yiyaodaobao.com.cn/CN/ 或 http://www.yiyaodaobao.com.cn/CN/Y2019/V16/I25/11

[1] Le Maitre CL，Pockert A，Buttle DJ，et al. Matrix synthesis and degradation in human intervertebral disc degeneration [J]. Biochem Soc Trans，2007，35（Pt 4）：652-655.
[2] Ding F，Shao ZW，Xiong LM. Cell death in intervertebral disc degeneration [J]. Apoptosis，2013，18（7）：777-785.
[3] Hwang PY，Chen J，Jing L，et al. The role of extracellular matrix elasticity and composition in regulating the nucleus pulposus cell phenotype in the intervertebral disc：a narrative review [J]. J Biomech Eng，2014，136（2）：021010.
[4] Chainani-Wu N. Safety and Anti-Inflammatory Activity of Curcumin：A Component of Tumeric （Curcuma longa） [J]. J Altern Complement Med，2003，9（1）：161-168.
[5] 周小园，容松，田梅.姜黄素对肾缺血再灌注损伤的保护作用机制的研究进展[J].中国医药导报，2019，16（6）：20-27.
[6] Rao CV. Regulation of cox and lox by curcumin [J]. Adv Exp Med Biol，2007，595（1）：213-226.
[7] Dinkova-Kostova AT，Talalay P. Direct and indirect antioxidant properties of inducers of cytoprotective proteins [J]. Mol Nutr Food Res，2008，52（S1）：S128-S138.
[8] Li X，Feng K，Li J，et al. Curcumin Inhibits Apoptosis of Chondrocytes through Activation ERK1/2 Signaling Pathways Induced Autophagy [J]. Nutrients，2017，9（4）：414.
[9] Wang J，Ma J，Gu JH，et al. Regulation of type II collagen，matrix metalloproteinase -13 and cell proliferation by interleukin-1β is mediated by curcumin via inhibition of NF-κB signaling in rat chondrocytes [J]. Mol Med Rep，2017，16（2）：1837-1845.
[10] Pfirrmann CW，Metzdorf A，Zanetti M，et al. Magnetic Resonance Classification of Lumbar Intervertebral Disc Degeneration [J]. Spine，2001，26（17）：1873-1878.
[11] Zhao CQ，Wang LM，Jiang LS，et al. The cell biology of intervertebral disc aging and degeneration [J]. Ageing Res Rev，2007，6（3）：247-261.
[12] Priyadarshani P，Li Y，Yao L. Advances in biological therapy for nucleus pulposus regeneration [J]. Osteoarthritis Cartilage，2016，24（2）：206-212.
[13] Le Maitre CL，Binch AL，Thorpe AA et al. Degeneration of the intervertebral disc with new approaches for treating low back pain [J]. J Neurosurg Sci，2015，59（1）：47-61.
[14] Shen J，Fang J，Hao J，et al. SIRT1 Inhibits the Catabolic Effect of IL-1β through TLR2/SIRT1/NF-κB Pathway in Human Degenerative Nucleus Pulposus Cells [J]. Pain Physician，2016，19（1）：E215.
[15] Wongcharoen W，Phrommintikul A. The protective role of curcumin in cardiovascular diseases [J]. Int J Cardiol，2009，133（2）：145-151.
[16] Mathy-Hartert M，Jacquemond-Collet I，Priem F，et al. Curcumin inhibits pro-inflammatory mediators and metalloproteinase-3 production by chondrocytes [J]. Inflamm Res，2009，58（12）：899-908.
[17] 张锐，刘世清.姜黄素保护关节软骨抑制骨关节炎的作用和机制[J].中国组织工程研究，2015，19（2）：277-282.
[18] Mistiaen WP，Somers P，Knaapen MW，et al. Autophagy as mechanism for cell death in degenerative aortic valve disease [J]. Autophagy，2006，2（3）：221-223.
[19] Bredesen DE，Rao RV，Mehlen P. Cell death in the nervous system [J]. Nature，2006，443（7113）：796-802.
[20] Jiang W，Zhang X，Hao J，et al. SIRT1 protects against apoptosis by promoting autophagy in degenerative human disc nucleus pulposus cells [J]. Sci Rep，2014，4：7456.