Resveratrol alleviates the interleukin-1β-induced chondrocytes injury through the NF-κB signaling pathway
Authors: Xin-Hui Zhu
Jian-Bo Fan
Wei Liu
Zhi-Ming Cui
Wei Zhang
Hong Yi
Sheng-Yu Cui
Journal: Journal of Orthopaedic Surgery and Research (2020)
Discipline: CLINICAL MEDICINE
Keywords: MMP-1; MMP-13; MMP-3; NF-κB signaling pathway; Osteoarthritis
Affiliations: Department of Orthopedic, the Affiliated Hospital 2 of Nantong university & the First People's Hospital of Nantong, No. 6 Haierxiangbei Road, Nantong City, 226001, Jiangsu Province, China., Department of Orthopedic, the Affiliated Hospital 2 of Nantong university & the First People's Hospital of Nantong, No. 6 Haierxiangbei Road, Nantong City, 226001, Jiangsu Province, China. .

Abstract:  Background: Osteoarthritis (OA) is a regular age-related disease that affects millions of people. Resveratrol (RSV) is a flavonoid with a stilbene structure with different pharmacological effects. The purpose of the experiment was to evaluate the protective role of RSV against the human OA chondrocyte injury induced by interleukin-1β (IL-1β). Methods: Chondrocytes were isolated from OA patients and identified by type II collagen, safranin O staining, and toluidine blue staining. Differentially expressed genes in chondrocytes treated RSV were identified by RNA sequencing. Kyoto encyclopedia of genes and genomes (KEGG) pathway as well as gene ontology (GO) were further conducted through Metascape online tool. A cell counting kit-8 (CCK-8) assay was applied to discover the viability of chondrocytes (6, 12, 24, and 48 μM). Many genes associated with inflammation and matrix degradation are evaluated by real-time PCR (RT-PCR) as well as western blot (WB). The mechanism of RSV for protecting IL-1β induced chondrocytes injury was further measured through immunofluorescence and WB assays. Results: A total of 845 differentially expressed genes (upregulated = 499, downregulated = 346) were found. These differentially expressed genes mainly enriched into negative regulation of catabolic process, autophagy, and cellular catabolic process, intrinsic apoptotic, apoptotic, and regulation of apoptotic signaling pathway, cellular response to abiotic stimulus, external stimuli, stress, and radiation. These differentially expressed genes were obviously enriched in NF-kB signaling pathway. RSV at the concentration of 48 μM markedly weakened the viability of the cells after 24 h of treatment (87% vs 100%, P &lt; 0.05). No obvious difference was observed between the 6, 12, and 24 μM groups (106% vs 100%, 104% vs 100%, 103% vs 100%, P &gt; 0.05). RSV (24 μM) also markedly depressed the levels of PGE2 and NO induced by IL-1β by 25% and 29% respectively (P &lt; 0.05). Our experiment pointed out that RSV could dramatically inhibit the inflammatory response induced by IL-1β, including the MMP-13, MMP-3, and MMP-1 in human OA chondrocytes by 50%, 35%, and 33% respectively. On the other hand, RSV inhibited cyclooxygenase-2 (COX-2), matrix metalloproteinase-1 (MMP-1), MMP-3, MMP-13, and inducible nitric oxide synthase (iNOs) expression (P &lt; 0.05), while increased collagen-II and aggrecan levels (P &lt; 0.05). From a mechanistic perspective, RSV inhibited the degradation of IκB-α as well as the activation of nuclear factor-kappa B (NF-κB) induced by IL-1β. Conclusion: In summary, RSV regulates the signaling pathway of NF-κB, thus inhibiting inflammation and matrix degradation in chondrocytes. More studies should be focused on the treatment efficacy of RSV for OA in vivo. 

Retracted: No

PubPeer comments:
 The Resveratrol and Control sample images appear to overlap. DAPI images marked, but this appears to occur in all 3 sets of images. Apologies to a Dr. Wei Zhang who I mistakenly included via email. This was a researcher with the same name incorrectly listed as an author on this paper at Researchgate. The moderators will hopefully remove the incorrect address. *** ![file] ,