lithium-chloride and Osteoarthritis

Nowadays, the cumulative intake of glucocorticoids has become the most common pathogenic factor for non-traumatic osteonecrosis of the femoral head (ONFH). Apoptosis of osteoblasts is considered as the main reason of ONFH at the molecular level. Glycogen synthase kinase 3β (GSK3β) is an important regulator of cellular differentiation and apoptosis pathway, which can modulate the balance between osteoblasts and osteoclasts. Several studies have reported about its function in osteoporosis, but little is known about it in osteonecrosis. In our study, lipopolysaccharide and methylprednisolone were utilized to establish a rat ONFH model. The phosphorylation of GSK3β Ser-9 was decreased in the model. Western blotting examination of β-catenin, Bcl-2, Bax and caspase-3 revealed that the osteoblasts were apoptotic. In dexamethasone (Dex)-incubated primary osteoblasts, the expression profile of GSK3β phosphorylation and apoptotic factors were consistent with those in the rat ONFH model. To further investigate the regulation of osteonecrosis caused by GSK3β, the expression and function of GSK3β were inhibited in Dex-incubated primary osteoblasts. The knockdown of GSK3β by siRNA decreased the expression of Bax and cleaved caspase-3, but increased Bcl-2 and β-catenin. On the other hand, selective inhibition of GSK3β function by LiCl counteracted the activation of caspase-3 induced by Dex. Our work is the first study about the GSK3β phosphorylation in ONFH, and provides evidence for further therapeutic methods.

Topics: Animals; Apoptosis; Dexamethasone; Disease Models, Animal; Femur Head Necrosis; Gene Knockdown Techniques; Glycogen Synthase Kinase 3 beta; Lipopolysaccharides; Lithium Chloride; Methylprednisolone; Osteoarthritis; Osteoblasts; Phosphorylation; Rats; Serine; Up-Regulation

Abnormal activation of the Wnt/β-catenin signaling is implicated in the osteoarthritis (OA) pathology. We searched for a pre-approved drug that suppresses abnormally activated Wnt/β-catenin signaling and has a potency to reduce joint pathology in OA. We introduced the TOPFlash reporter plasmid into HCS-2/8 human chondrosarcoma cells to estimate the Wnt/β-catenin activity in the presence of 10 μM each compound in a panel of pre-approved drugs. We found that fluoxetine, an antidepressant in the class of selective serotonin reuptake inhibitors (SSRI), down-regulated Wnt/β-catenin signaling in human chondrosarcoma cells. Fluoxetine inhibited both Wnt3A- and LiCl-induced loss of proteoglycans in chondrogenically differentiated ATDC5 cells. Fluoxetine increased expression of Sox9 (the chondrogenic master regulator), and decreased expressions of Axin2 (a marker for Wnt/β-catenin signaling) and Mmp13 (matrix metalloproteinase 13). Fluoxetine suppressed a LiCl-induced increase of total β-catenin and a LiCl-induced decrease of phosphorylated β-catenin in a dose-dependent manner. An in vitro protein-binding assay showed that fluoxetine enhanced binding of β-catenin with Axin1, which is a scaffold protein forming the degradation complex for β-catenin. Fluoxetine suppressed LiCl-induced β-catenin accumulation in human OA chondrocytes. Intraarticular injection of fluoxetine in a rat OA model ameliorated OA progression and suppressed β-catenin accumulation.

Topics: Animals; Axin Protein; Cartilage, Articular; Cell Differentiation; Cells, Cultured; Chondrocytes; Chondrogenesis; Disease Models, Animal; Down-Regulation; Fluoxetine; Humans; Lithium Chloride; Matrix Metalloproteinase 13; Osteoarthritis; Phosphorylation; Rats; Rats, Sprague-Dawley; Selective Serotonin Reuptake Inhibitors; SOX9 Transcription Factor; Wnt Signaling Pathway

The present study aimed to investigate the effect of palmatine (Pal) in a rabbit osteoarthritis (OA) model in vivo and rabbit interleukin-1β (IL-1β)-stimulated chondrocytes in vitro. Appropriate concentrations of Pal were identified by the MTT assay and used to preincubate IL-1β-induced chondrocytes, as well as an activator or inhibitor of Wnt and Hedgehog signaling pathways. Matrix metalloproteinase (MMP)-1, 3, and 13; tissue inhibitor of metalloproteinase (TIMP)-1; collagenase II; aggrecan; and the related molecules of the Wnt/β-catenin and Hedgehog signaling pathways were investigated. Protein expression was detected by Western blot analysis and messenger RNA (mRNA) expression was examined by PCR analysis. Pal (0.3 mL, 100 mg/L) was injected into rabbit knee joints and histological examination, immunohistochemistry, and Mankin scoring of the articular cartilage were performed. Pal (10-100 mg/L) had no effect on chondrocyte viability, decreased the expression of the MMPs, and increased the synthesis of TIMP-1whereas collagenase II and aggrecan were inhibited by IL-1β. When the activator (Licl) and inhibitor (DKK-1) of the Wnt/β-catenin signaling pathway as well as the inhibitor (cyclopamine) of the Hedgehog signaling pathway were added, the Wnt/β-catenin signaling pathway was less inhibited by Pal, and a similar inhibitory effect of cyclopamine on the Hedgehog signaling pathway was evident. Additionally, Pal enhanced the effect of cyclopamine. The histological examination, immunohistochemistry and Mankin scoring also demonstrated the protective effect of Pal, and the inhibition of the Wnt and Hedgehog signaling pathways by Pal. Pal may be useful in the treatment of OA, in which its effect is likely mediated via the Wnt/β-catenin and Hedgehog signaling pathways.

Topics: Animals; Berberine Alkaloids; beta Catenin; Cells, Cultured; Chondrocytes; Drug Synergism; Hedgehogs; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-1beta; Lithium Chloride; Matrix Metalloproteinases; Osteoarthritis; Rabbits; Tissue Inhibitor of Metalloproteinase-1; Veratrum; Veratrum Alkaloids; Wnt Proteins; Wnt Signaling Pathway

Molecular hydrogen (H2) is effective for many diseases. However, molecular bases of H2 have not been fully elucidated. Cumulative evidence indicates that H2 acts as a gaseous signal modulator. We found that H2 suppresses activated Wnt/β-catenin signaling by promoting phosphorylation and degradation οf β-catenin. Either complete inhibition of GSK3 or mutations at CK1- and GSK3-phosphorylation sites of β-catenin abolished the suppressive effect of H2. H2 did not increase GSK3-mediated phosphorylation of glycogen synthase, indicating that H2 has no direct effect on GSK3 itself. Knock-down of adenomatous polyposis coli (APC) or Axin1, which form the β-catenin degradation complex, minimized the suppressive effect of H2 on β-catenin accumulation. Accordingly, the effect of H2 requires CK1/GSK3-phosphorylation sites of β-catenin, as well as the β-catenin degradation complex comprised of CK1, GSK3, APC, and Axin1. We additionally found that H2 reduces the activation of Wnt/β-catenin signaling in human osteoarthritis chondrocytes. Oral intake of H2 water tended to ameliorate cartilage degradation in a surgery-induced rat osteoarthritis model through attenuating β-catenin accumulation. We first demonstrate that H2 suppresses abnormally activated Wnt/β-catenin signaling, which accounts for the protective roles of H2 in a fraction of diseases.

Topics: Adenomatous Polyposis Coli Protein; Animals; Axin Protein; beta Catenin; Casein Kinase I; Cell Line; Chondrocytes; Gases; Glycogen Synthase Kinase 3; HCT116 Cells; HeLa Cells; HT29 Cells; Humans; Hydrogen; Leupeptins; Lithium Chloride; Male; Microscopy, Fluorescence; Osteoarthritis; Phosphorylation; Rats; Rats, Sprague-Dawley; RNA Interference; RNA, Small Interfering; SOX9 Transcription Factor; Water; Wnt Signaling Pathway; Wnt3A Protein

Osteoarthritis is a chronic degenerative disease that affects the articular cartilage. Recent studies have demonstrated that lithium chloride exhibits significant efficacy as a chondroprotective agent, blocking cartilage degradation in response to inflammatory cytokines. However, conflicting literature suggests lithium may affect the physicochemical properties of articular cartilage and thus long-term exposure may negatively affect the mechanical functionality of this tissue. This study aims to investigate the effect of lithium chloride on the biomechanical properties of healthy and interleukin-1β treated cartilage in vitro and examines the consequences of long-term exposure to lithium on cartilage health in vivo. Bovine cartilage explants were treated with lithium chloride for 12 days. Chondrocyte viability, matrix catabolism and the biomechanical properties of bovine cartilage explants were not significantly altered following treatment. Consistent with these findings, long term-exposure (9 months) to dietary lithium did not induce osteoarthritis in rats, as determined by histological staining. Moreover, lithium chloride did not induce the expression of catabolic enzymes in human articular chondrocytes. In an inflammatory model of cartilage destruction, lithium chloride blocked interleukin-1β signaling in the form of nitric oxide and prostaglandin E2 release and prevented matrix catabolism such that the loss of mechanical integrity observed with interleukin-1β alone was inhibited. This study provides further support for lithium chloride as a novel compound for the treatment of osteoarthritis.

Topics: Animals; Cartilage, Articular; Cattle; Cells, Cultured; Drug Evaluation, Preclinical; Humans; Interleukin-1beta; Lithium Chloride; Male; Osteoarthritis; Rats, Wistar

To determine the actions of lithium chloride (LiCl) on catabolic events in human articular chondrocytes, and the effects of LiCl on the progression and severity of cartilage degradation in interleukin-1β (IL-1β)-treated mouse knee joints and after surgical induction of osteoarthritis (OA) in a mouse model.. Human articular chondrocytes were treated with LiCl followed by IL-1β, and the expression levels of catabolic genes were determined by real-time polymerase chain reaction. To understand the mechanism by which LiCl affects catabolic events in articular chondrocytes after IL-1β treatment, the activation of NF-κB was determined using luciferase reporter assays, and the activities of MAPKs and the STAT-3 signaling pathway were determined by immunoblot analysis of total cell lysates. Cultures of mouse femoral head explants treated with IL-1β and a mouse model of surgically induced OA were used to determine the effects of LiCl on proteoglycan loss and cartilage degradation.. LiCl treatment resulted in decreased catabolic marker messenger RNA levels and activation of NF-κB, p38 MAPK, and STAT-3 signaling in IL-1β-treated articular chondrocytes. Furthermore, LiCl directly inhibited IL-6-stimulated activation of STAT-3 signaling. Consequently, the loss of proteoglycan and severity of cartilage destruction in LiCl-treated mouse knee joints 8 weeks after OA induction surgery or in LiCl-treated mouse femoral head explants after IL-1β treatment were markedly reduced compared to that in vehicle-treated joints or explants.. LiCl reduced catabolic events in IL-1β-treated human articular chondrocytes and attenuated the severity of cartilage destruction in IL-1β-treated mouse femoral head explants and in the knee joints of mice with surgically induced OA, acting via inhibition of the activities of the NF-κB, p38, and STAT-3 signaling pathways.

Topics: Aged; Animals; Cartilage, Articular; Cells, Cultured; Chondrocytes; Disease Models, Animal; Disease Progression; Femur Head; Humans; Interleukin-1beta; Lithium Chloride; Male; Mice; Mice, Inbred C57BL; Middle Aged; NF-kappa B; Osteoarthritis; p38 Mitogen-Activated Protein Kinases; Proteoglycans; Severity of Illness Index; Signal Transduction; STAT3 Transcription Factor

To determine the effects and mechanism of action of lithium chloride (LiCl) on cartilage destruction induced by the pro-inflammatory cytokines IL-1, IL-1 + oncostatin M and TNF-α.. The release of collagen was assessed in bovine cartilage explant cultures, whereas collagenolytic activities (active and total) in conditioned culture supernatants were determined by bioassay. The expression and production of MMP from chondrocytes were analysed by real-time RT-PCR and ELISA. Signalling pathway analysis was performed using a phospho-antibody array and standard immunoblotting.. LiCl, but not selective glycogen synthase kinase 3 (GSK-3) inhibitor compounds SB-415286 and TDZD-8, significantly decreased pro-inflammatory cytokine-induced collagen release from bovine cartilage via the down-regulation of collagenolytic activity. Furthermore, MMP-1 and MMP-13 expression was reduced in both bovine and human chondrocytes. Pathway analysis revealed that LiCl selectively inhibited activation of the p38 mitogen-activated protein kinase pathway; effects that were recapitulated by specific p38 pathway inhibition.. This study demonstrates for the first time that LiCl can protect against cartilage damage induced by pro-inflammatory cytokines, and indicates that LiCl-mediated cartilage protection is not via a GSK-3-dependent mechanism, but potentially via inhibition of the p38 pathway. These data indicate that lithium administration may represent a potential therapy for arthritis.

Topics: Aged; Animals; Cartilage; Cattle; Chondrocytes; Down-Regulation; Enzyme-Linked Immunosorbent Assay; Humans; Lithium Chloride; Matrix Metalloproteinases; Osteoarthritis; p38 Mitogen-Activated Protein Kinases; Reverse Transcriptase Polymerase Chain Reaction

Topics: Adult; Aged; Arthritis, Rheumatoid; Chlorides; Drug Evaluation; Drug Therapy, Combination; Female; Humans; Iontophoresis; Joints; Lithium; Lithium Chloride; Male; Middle Aged; Osteoarthritis; Radionuclide Imaging; Time Factors