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Effects of curcumin on autophagy and degeneration of human intervertebral disc nucleus pulposus |
WAN Junfeng YANG Bo ZHENG Jiazhuang RAN Maobo SONG Zhaojun |
Department of Orthopedics, Suining Central Hospital, Sichuan Province, Suining 629000, China |
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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.
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