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Dexmedetomidine (Dex) is an anesthetic widely used in lumbar discectomy, but its effect on chondrocytes remains unclear. Dex is speculated to promote cartilage degeneration by activating α-2 adrenergic receptor. However, the antioxidative and anti-inflammatory effects of Dex implied the potential chondrocyte protective effect under stress conditions. The present study aimed to determine the effect of Dex on chondrocytes under non-stress and stress conditions. Chondrocytes were isolated from human annulus fibrosus (AF) tissues and oxidative stress was induced by treatment with 1 mM hydrogen peroxide (H2O2). Chondrocytes were treated with Dex alone or in combination with H2O2 Treatment with Dex alone decreased mRNA expression of COL2A1 and increased that of MMP-3 and MMP-13, thus contributing to cartilage degeneration. However, Dex prevented H2O2-induced death and degeneration of chondrocytes partly by enhancing antioxidant capacity. Mechanistically, Dex attenuated H2O2-mediated activation of NF-κB and NACHT, LRR, and PYD domains-containing protein 3 (NLRP3), both of which play key roles in inflammation and inflammatory damage. Dex inactivated NLRP3 through the suppression of NF-κB and JNK signals. Co-treatment with Dex and H2O2 increased protein level of XIAP (X-linked inhibitor-of-apoptosis, an anti-apoptosis protein), compared with H2O2 treatment alone. H2O2 treatment increased the expression of neural precursor cell expressed developmentally down-regulated protein 4 (NEDD4) that is a ubiquitin ligase targeting XIAP. However, Dex decreased the amount of NEDD4 adhering to XIAP, thus protecting XIAP protein from NEDD4-mediated ubiquitination and degradation. Given that surgery inevitably causes oxidative stress and inflammation, the protective effect of Dex on chondrocytes during oxidative stress is noteworthy and warrants further study.

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Dysfunction of cartilaginous endplates (CEP) is an important etiologic aspect of intervertebral disc degeneration (IDD) because the endplate has nutritional and biomechanical functions in maintaining proper disc health. In this study, we investigated the regulatory effects of estrogen on degenerated human CEP cells and the involvement of miR-221 in these effects. Normal and degenerated human CEP tissues were collected from patients with idiopathic scoliosis and IDD, respectively. CEP cells were isolated from these tissues. Polymerase chain reaction (PCR) and western blot analysis were performed to detect the expression of specific genes and proteins, respectively. Apoptosis and cell cycle were analyzed by flow cytometry. The results showed that the levels of aggrecan, collagen II, TGF-ß and estrogen receptor α (ERα) were decreased in degenerated CEP tissues, while the levels of MMP-3, adamts-5, IL-1ß, TNF-α, IL-6, and miR-221 were increased. Treatment of degenerated CEP cells with 17beta-estradiol (E2) increased the expressions of aggrecan and collagen II, as well as the secretion of TGF-ß, but decreased IL-6 secretion. Moreover, E2 inhibited the apoptosis, resumed cell-cycle progression in G0/G1 phase, and improved the cell viability. These data indicate that estrogen has protective effect against degeneration of CEP cells. Furthermore, ERα was confirmed to be a target of miR-221 by the luciferase assay. The synthetic miR-221 mimics or knockdown of ERα attenuated the protective effects of E2, but miR-221 inhibitors promoted the protective effects of E2. These results suggest that miR-221 may impair the protective effect of estrogen in degenerated CEP cells through targeting ERα. This study reveals an important mechanism underlying the degeneration of CEP cells.

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Clinical studies report that endogenous estrogen depletion is associated with disc degeneration. The present study aimed to investigate the effect and mechanism of estrogen on disc degeneration of the cartilage endplate. Three groups of mice with bilateral ovariectomy + 17ß-estradiol injection (OVX + E2 Group), bilateral ovariectomy + vehicle injection (OVX + vehicle), or sham operation + vehicle injection (Sham Group) were included in this study. The mice were sacrificed at 12 weeks and the cartilage endplate (CEP) were harvested. The calcification status was evaluated by Alizarin red staining and RT-PCR, which demonstrated the calcification level of the CEP gradually developed from the Sham Group, OVX + E2, to the OVX + vehicle group. The CEP cells were isolated, cultured, and treated with IL-1ß (75 ng/ml) for 24 h, with or without a pretreatment of 17ß-E2 for 1 h. RT-PCR analysis of calcification-related genes ALP, OCN, RUNX2, and COL-I were analyzed, and calcification of CEP cells induced by IL-1ß was reversed by pretreatment with 17ß-E2, in a dose-dependent manner. The protective effect of 17ß-E2 was abolished by estrogen receptor antagonist ICI182,780. These results suggest that decreased estrogen level may accelerate degeneration of the cartilage endplate by increasing calcification, which may be induced by IL-1ß, in a dose-dependent manner.