Effects of curcumin on apoptosis and proliferation of inflammatory chondrocytes induced by lipopolysaccharide

doi:10.3969/j.issn.2095-4344.0539

Abstract

Abstract:

BACKGROUND: Curcumin has been shown to play antioxidative, anti-inflammatory and anti-apoptotic roles in osteoarthritis, but the underlying mechanisms remain unclear.
OBJECTIVE: To investigate the effects of curcumin on apoptosis and proliferation of chondrocytes induced by lipopolysaccharide, and to explore the potential mechanisms, so as to provide new ideas for the treatment of osteoarthritis.
METHODS: Chondrocytes were isolated from the 4-week-old Sprague-Dawley rats, cultured, and identified by toluidine blue staining. The chondrocyte inflammation model in vitro was established by lipopolysaccharide induction. The cells were divided into five groups: control group (common medium); model group (medium with 5 µg/L lipopolysaccharide); low-dose group (medium with 5 µg/L lipopolysaccharide + 5 µmol/L curcumin); medium-dose group (medium with 5 µg/L lipopolysaccharide + 10 µmol/L curcumin); high-dose group (medium with 5 µg/L lipopolysaccharide + 20 µmol/L curcumin). After 24 hours, the expression levels of interleukin 1β and transforming growth factor α in the cellular supernatant were detected by ELISA method. The apoptotic cells were detected by flow cytometry. Cell proliferation was tested by MTT assay. The expression levels of autophagy-related proteins and extracellular regulated protein kinases 1/2 were determined by western blot assay.
RESULTS AND CONCLUSION: The levels of interleukin 1β and transforming growth factor α in the supernatant of chondrocytes induced by lipopolysaccharide were significantly higher than those in the control group (P < 0.01). The levels of interleukin 1β and transforming growth factor α in the supernatant treated with curcumin were lower than those in the model group, which was on a dose-dependent manner (P < 0.05, P < 0.01). After 24 hours of treatment, apoptotic rate of cells in the model group was significantly higher than that in the control group (P < 0.01). Compared with the model group, the cell apoptotic rate in each curcumin group was decreased, especially the high-dose group (P < 0.01). MTT assay showed that the cell proliferation ability in the model group was significantly lower than that in the control group (P < 0.01). Compared with the model group, the cell proliferation ability in each curcumin group was restored, and the effect was the best in the high-dose group. The results of western blot showed that the autophagic ability and the levels of autophagy marker (LC3-II) and autophagy related protein (Beclin-1) were significantly increased in each curcumin group compared with the model group(P < 0.05, P < 0.01), and the expression level of p-ERK1/2 protein was significantly increased (P < 0.05). Our findings suggest that 20 µmol/L curcumin cannot only inhibit apoptosis and relieve inflammatory of chondrocytes induced by lipopolysaccharide, but also has a certain antagonistic effect on lipopolysaccharide- induced inhibition of inflammatory chondrocyte proliferation, which may be via regulating the ERK1/2 pathway-activated autophagy.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Osteoarthritis, Curcumin, Cell Proliferation, Apoptosis, Tissue Engineering

CLC Number:

R318

Li Ya-nan, Ni Juan, Fang Yu-shun, Li Tao, Tan Hong-fei, Zhang Qing-song . Effects of curcumin on apoptosis and proliferation of inflammatory chondrocytes induced by lipopolysaccharide[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(32): 5157-5162.

Figures/Tables 2

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