lignans and Intervertebral-Disc-Degeneration

Arctigenin (ATG) is the active ingredient of the Chinese herbal medicine Arctium lappa, with anti-inflammatory and antioxidant effects. Excessive inflammation and cell apoptosis are important causes of intervertebral disc degeneration (IDD). Hence, this study probed into the possible role of ATG in IDD.. Interleukin (IL)-1β (10 ng/ml) was adopted to induce human nucleus pulposus cells (HNPCs) as a cell model for IDD. The effects of different concentrations of ATG (0, 2, 5, 10, 20, 50 μmol/L) on the viability of HNPCs and effects of ATG (10, 50 μmol/L) on the viability of IL-1β-induced HNPCs were detected by cell counting kit-8 (CCK-8). After IL-1β-induced HNPCs were transfected with miR-483-3p inhibitor and/or treated with ATG, cell viability and apoptosis were determined by CCK-8 and flow cytometry; the expressions of miR-483-3p, extracellular matrix (ECM)-related genes, and inflammation-related genes were measured by quantitative real time polymerase chain reaction (qRT-PCR), and expressions of ECM/apoptosis/NF-κB pathway-related proteins were quantified by Western blot.. ATG had no significant effect on the viability of HNPCs but could promote the viability of IL-1β-induced HNPCs. ATG inhibited apoptosis, ECM degradation, inflammation, and activation of NF-κB pathway in HNPCs induced by IL-1β, but promoted the expression of miR-483-3p. MiR-483-3p inhibitor reversed the above-mentioned regulatory effects of ATG.. Arctigenin suppresses apoptosis, ECM degradation, inflammation, and NF-κB pathway activation in HNPCs by up-regulating miR-483-3p.

Topics: Apoptosis; Cells, Cultured; Extracellular Matrix; Extracellular Matrix Proteins; Furans; Humans; Inflammation; Intervertebral Disc Degeneration; Lignans; MicroRNAs; NF-kappa B; Nucleus Pulposus

Intervertebral disc degeneration-related diseases are common health problems in the department of orthopedics. However, there is no effective treatment protecting the intervertebral disc from degeneration. Sesamin, a kind of sesame lignans extracted from sesame seed oil, has been proved to inhibit lipopolysaccharide-induced inflammation and extracellular matrix catabolism in rat intervertebral disc in vitro and ex vivo. The present study was designed to investigate the effects of sesamin on lesion-induced intervertebral disc degeneration in vivo. Degeneration of rat tail disc was induced by puncture lesion, followed by intradiscal injection of sesamin. Magnetic resonance imaging (MRI), quantitative real-time polymerase chain reaction, histological analysis, and biochemical analysis were carried out to analyze degeneration progression 2 weeks after surgery. As shown by results, intradiscal injection of sesamin inhibited the MRI signal decrease of nucleus pulposus (NP) in T2-weighted images. The upregulated mRNA expression of MMP-3 and ADAMTS-5 induced by lesion was significantly suppressed by sesamin injection. Sesamin partly protected mRNA expression of Col2a1 and Acan from downregulation. Intradiscal injection of sesamin effectively maintained the normal morphology of disc and inhibited lesion-induced degeneration-related histological changes. Immunohistochemical assay demonstrated that the upregulation of degradative enzymes protein expression and the downregulation of type II collagen expression in NP were suppressed by sesamin. According to biochemical analysis, sesamin significantly inhibited the lesion-induced decrease of proteoglycan content in NP. The present study proved the protective effects of sesamin on lesion-induced intervertebral disc degeneration at an early stage.

Topics: Animals; Dioxoles; Disease Models, Animal; DNA; Extracellular Matrix; Gene Expression Regulation; Intervertebral Disc; Intervertebral Disc Degeneration; Lignans; Magnetic Resonance Imaging; Nucleus Pulposus; Proteoglycans; Rats, Sprague-Dawley; RNA, Messenger

Topics: Animals; Antioxidants; Biphenyl Compounds; Carrier Proteins; Cell Cycle Proteins; Inflammasomes; Intervertebral Disc Degeneration; Lignans; Male; NLR Family, Pyrin Domain-Containing 3 Protein; Nucleus Pulposus; Rats; Rats, Sprague-Dawley; Signal Transduction

Oxidative stress-induced mitochondrial dysfunction is implicated in the pathogenesis of intervertebral disc degeneration (IVDD). Sirtuin 3 (SIRT3), a sirtuin family protein located in mitochondria, is essential for mitochondrial homeostasis; however, the role of SIRT3 in the process of IVDD has remained elusive. Here, we explored the expression of SIRT3 in IVDD in vivo and in vitro; we also explored the role of SIRT3 in senescence, apoptosis, and mitochondrial homeostasis under oxidative stress. We subsequently activated SIRT3 using honokiol to evaluate its therapeutic potential for IVDD. We assessed SIRT3 expression in degenerative nucleus pulposus (NP) tissues and oxidative stress-induced nucleus pulposus cells (NPCs). SIRT3 was knocked down by lentivirus and activated by honokiol to determine its role in oxidative stress-induced NPCs. The mechanism by which honokiol affected SIRT3 regulation was investigated in vitro, and the therapeutic potential of honokiol was assessed in vitro and in vivo. We found that the expression of SIRT3 decreased with IVDD, and SIRT3 knockdown reduced the tolerance of NPCs to oxidative stress. Honokiol (10 μM) improved the viability of NPCs under oxidative stress and promoted their properties of anti-oxidation, mitochondrial dynamics and mitophagy in a SIRT3-dependent manner. Furthermore, honokiol activated SIRT3 through the AMPK-PGC-1α signaling pathway. Moreover, honokiol treatment ameliorated IVDD in rats. Our study indicated that SIRT3 is involved in IVDD and showed the potential of the SIRT3 agonist honokiol for the treatment of IVDD.

Topics: Animals; Antioxidants; Biphenyl Compounds; Cells, Cultured; Female; Humans; Intervertebral Disc Degeneration; Lignans; Male; Middle Aged; Mitochondrial Dynamics; Nucleus Pulposus; Oxidative Stress; Rats; Rats, Sprague-Dawley; Sirtuin 3

Intervertebral disc (IVD) degeneration contributes to most spinal degenerative diseases, while treatment inhibiting IVD degeneration is still in the experimental stage. Sesamin, a bioactive component extracted from sesame, has been reported to exert chondroprotective and anti-inflammatory effects. Here, we analyzed the anti-inflammatory and anti-catabolic effects of sesamin on rat IVD in vitro and ex vivo. Results show that sesamin significantly inhibits the lipopolysaccharide (LPS)-induced expression of catabolic enzymes (MMP-1, MMP-3, MMP-13, ADAMTS-4, ADAMTS-5) and inflammation factors (IL-1β, TNF-α, iNOS, NO, COX-2, PGE2) in a dose-dependent manner in vitro. It is also proven that migration of macrophages induced by LPS can be inhibited by treatment with sesamin. Organ culture experiments demonstrate that sesamin protects the IVD from LPS-induced depletion of the extracellular matrix ex vivo. Moreover, sesamin suppresses LPS-induced activation of the mitogen-activated protein kinase (MAPK) pathway through inhibiting phosphorylation of JNK, the common downstream signaling pathway of LPS and IL-1β, which may be the potential mechanism of the effects of sesamin. In light of our results, sesamin protects the IVD from inflammation and extracellular matrix catabolism, presenting positive prospects in the treatment of IVD degenerative diseases.

Topics: Animals; Cell Movement; Cell Survival; Cells, Cultured; Cyclooxygenase 2; Dinoprostone; Dioxoles; Enzyme Activation; Extracellular Matrix; Inflammation; Interleukin-1beta; Intervertebral Disc; Intervertebral Disc Degeneration; JNK Mitogen-Activated Protein Kinases; Lignans; Lipopolysaccharides; Macrophages; MAP Kinase Signaling System; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Nucleus Pulposus; Rats, Sprague-Dawley; Real-Time Polymerase Chain Reaction; RNA, Messenger; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha