The role of ferroptosis in intervertebral disc degeneration.

Nucleus pulposus, annulus fibrosus, and cartilage endplate constitute an avascular intervertebral disc (IVD), which is crucial for spinal and intervertebral joint mobility. As one of the most widespread health issues worldwide, intervertebral disc degeneration (IVDD) is recognized as a key contributor to back and neck discomfort. A number of degenerative disorders have a strong correlation with ferroptosis, a recently identified novel regulated cell death (RCD) characterized by an iron-dependent mechanism and a buildup of lipid reactive oxygen species (ROS). There is growing interest in the part ferroptosis plays in IVDD pathophysiology. Inhibiting ferroptosis has been shown to control IVDD development. Several studies have demonstrated that in TBHP-induced oxidative stress models, changes in ferroptosis marker protein levels and increased lipid peroxidation lead to the degeneration of intervertebral disc cells, which subsequently aggravates IVDD. Similarly, IVDD is significantly relieved with the use of ferroptosis inhibitors. The purpose of this review was threefold: 1) to discuss the occurrence of ferroptosis in IVDD; 2) to understand the mechanism of ferroptosis and its role in IVDD pathophysiology; and 3) to investigate the feasibility and prospect of ferroptosis in IVDD treatment.

A Minimally Invasive Annulus Fibrosus Needle Puncture Model of Intervertebral Disc Degeneration in Rats.

BACKGROUND: The needle puncture model in rats has been accepted as an ordinary model to induce intervertebral disc degeneration (IVDD). However, the model primarily penetrated the whole intervertebral disc, resulting in injury to the nucleus pulposus (NP) and annulus fibrosus (AF). The intention of this research was to explore a minimally invasive approach through needle puncture of the AF percutaneously in rats. METHODS: Twenty SD rats underwent puncture at Co8/9 via a 20 G percutaneous needle. The needle was slowly advanced perpendicular to the tail skin to penetrate the whole AF without damaging the NP limited by a hand-made sheath. The X-rays and magnetic resonance imaging T2 relaxation was evaluated at 1, 2, and 3 weeks to assess the disc height index and signal changes. Histological and immunohistochemical staining of the IVD were obtained under a light microscope. RESULTS: X-rays showed that the disc height had progressively narrowed to 49% of baseline 3 weeks after injury. magnetic resonance imaging evaluation demonstrated that the mean T2-weighted signal intensity at 3 weeks was 43% of that in the uninjured control group at the Co8/9 level. Histological staining demonstrated disorganized lamellae in the AF and decreased proteoglycan content and cellularity within the NP in the injured discs. CONCLUSIONS: The present research demonstrates a reliable and convenient approach to induce an AF tear in rats through percutaneous needle puncture. This model reduces harm to the experimental animals significantly while imitating the progressive degeneration process. More importantly, the model confirmed that AF damage alone could lead to IVDD and provided a research method for AF degeneration in IVDD.

CB2R Attenuates Intervertebral Disc Degeneration by Delaying Nucleus Pulposus Cell Senescence through AMPK/GSK3ß Pathway.

Nucleus pulposus (NP) cell (NPC) senescence is one of the main causes of intervertebral disc degeneration (IVDD). However, the underlying mechanism of NPC senescence is still unclear. The cannabinoid type 2 receptor (CB2R) is a member of the cannabinoid system and plays an important role in antioxidative stress, anti-inflammatory and antisenescence activities. In this study, we used a hydrogen peroxide (H2O2)-induced NPC senescence model and a rat acupuncture IVDD model to explore the role of CB2R in IVDD in vitro and in vivo. First, we confirmed that the expression of p16INK4a in the NP tissues of IVDD patients and rat acupuncture IVDD models obviously increased accompanied by a decrease in CB2R expression. Subsequently, we found that activation of CB2R significantly reduced the number of SA-ß-gal positive cells and suppressed the expression of p16INK4a and senescence-related secretory phenotypes [SASP, including matrix metalloproteinase 9 and 13 (MMP9, MMP13) and high mobility group protein b1 (HMGB1)]. In addition, activation of CB2R promoted the expression of collagen type II (Col-2) and SRY-Box transcription factor 9 (SOX9), inhibit the expression of collagen type X (Col-X), and restore the balance of extracellular matrix (ECM) metabolism. In addition, the AMPK/GSK3ß pathway was shown to play an important role in CB2R regulation of NPC senescence. Inhibition of AMPK expression reversed the effect of JWH015 (a CB2R agonist). Finally, we further demonstrated that in the rat IVDD model, the AMPK/GSK3ß pathway was involved in the regulation of CB2R on NPC senescence. In conclusion, our experimental results prove that CB2R plays an important role in NPC senescence. Activation of CB2R can delay NPC senescence, restore the balance of ECM metabolism, and attenuate IVDD.

Melatonin Attenuates Intervertebral Disk Degeneration via Maintaining Cartilaginous Endplate Integrity in Rats.

OBJECTIVE: The aim of this study is to verify whether melatonin (Mel) could mitigate intervertebral disk degeneration (IVDD) in rats and to investigate the potential mechanism of it. METHOD: A rat acupuncture model of IVDD was established with intraperitoneal injection of Mel. The effect of Mel on IVDD was analyzed via radiologic and histological evaluations. The specific Mel receptors were investigated in both the nucleus pulposus (NP) and cartilaginous endplates (EPs). In vitro, EP cartilaginous cells (EPCs) were treated by different concentrations of Mel under lipopolysaccharide (LPS) and Luzindole conditions. In addition, LPS-induced inflammatory response and matrix degradation following nuclear factor kappa-B (NF-κB) pathway activation were investigated to confirm the potential mechanism of Mel on EPCs. RESULTS: The percent disk height index (%DHI) and MRI signal decreased after initial puncture in the degeneration group compared with the control group, while Mel treatment protected disk height from decline and prevented the loss of water during the degeneration process. In the meantime, the histological staining of the Mel groups showed more integrity and well-ordered construction of the NP and EPs in both low and high concentration than that of the degeneration group. In addition, more deep-brown staining of type II collagen (Coll-II) was shown in the Mel groups compared with the degeneration group. Furthermore, in rat samples, immunohistochemical staining showed more positive cells of Mel receptors 1a and 1b in the EPs, instead of in the NP. Moreover, evident osteochondral lacuna formation was observed in rat EPs in the degeneration group; after Mel treatment, the osteochondral destruction alleviated accompanying fewer receptor activator for nuclear factor-κB ligand (RANKL) and tartrate-resistant acid phosphatase (TRAP)-stained positive cells expressed in the EPs. In vitro, Mel could promote the proliferation of EPCs, which protected EPCs from degeneration under LPS treatment. What is more, Mel downregulated the inflammatory response and matrix degradation of EPCs activated by NF-κB pathway through binding to its specific receptors. CONCLUSION: These results indicate that Mel protects the integrity of the EPs and attenuates IVDD by binding to the Mel receptors in the EPs. It may alleviate the inflammatory response and matrix degradation of EPCs activated by NF-κB pathway.

Author Correction: Social attraction in Drosophila is regulated by the mushroom body and serotonergic system.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Social attraction in Drosophila is regulated by the mushroom body and serotonergic system.

Sociality is among the most important motivators of human behaviour. However, the neural mechanisms determining levels of sociality are largely unknown, primarily due to a lack of suitable animal models. Here, we report the presence of a surprising degree of general sociality in Drosophila. A newly-developed paradigm to study social approach behaviour in flies reveal that social cues perceive through both vision and olfaction converged in a central brain region, the γ lobe of the mushroom body, which exhibite activation in response to social experience. The activity of these γ neurons control the motivational drive for social interaction. At the molecular level, the serotonergic system is critical for social affinity. These results demonstrate that Drosophila are highly sociable, providing a suitable model system for elucidating the mechanisms underlying the motivation for sociality.

Intelligence Algorithms for Protein Classification by Mass Spectrometry.

Mass spectrometry (MS) is an important technique in protein research. Effective classification methods by MS data could contribute to early and less-invasive diagnosis and also facilitate developments in the bioinformatics field. As MS data is featured by high dimension, appropriate methods which can effectively deal with the large amount of MS data have been widely studied. In this paper, the applications of methods based on intelligence algorithms have been investigated. Firstly, classification and biomarker analysis methods using typical machine learning approaches have been discussed. Then those are followed by the Ensemble strategy algorithms. Clearly, simple and basic machine learning algorithms hardly addressed the various needs of protein MS classification. Preprocessing algorithms have been also studied, as these methods are useful for feature selection or feature extraction to improve classification performance. Protein MS data growing with data volume becomes complicated and large; improvements in classification methods in terms of classifier selection and combinations of different algorithms and preprocessing algorithms are more emphasized in further work.

CO Oxidation by Group 3 Metal Monoxide Cations Supported on [Fe(CO)4 ]2.

Infrared photodissociation spectroscopy of mass-selected heteronuclear cluster anions in the form of OMFe(CO)5- (M=Sc, Y, La) indicates that all these anions involve an 18-electron [Fe(CO)4 ]2- building block that is bonded with the M center through two bridged carbonyl ligands. The OLaFe(CO)5- anion is determined to be a CO-tagged complex involving a [Fe(CO)4 ]2- [LaO]+ anion core. In contrast, the OYFe(CO)5- anion is characterized to have a [Fe(CO)4 ]2- [Y(η2 -CO2 )]+ structure involving a side-on bonded CO2 ligand. The CO-tagged complex and the [Fe(CO)4 ]2- [Sc(η2 -CO2 )]+ isomer co-exist for the OScFe(CO)5- anion. These observations indicate that both the ScO+ and YO+ cations supported on [Fe(CO)4 ]2- are able to oxidize CO to CO2 . Theoretical analyses show that [Fe(CO)4 ]2- coordination significantly weakens the MO+ bond and decreases the energy gap of the interacting valence orbitals between MO+ and CO, leading to the CO oxidation reactions being both thermodynamically exothermic and kinetically facile.

Infrared Photodissociation Spectroscopy of Heterodinuclear Iron-Zinc and Cobalt-Zinc Carbonyl Cation Complexes.

Fe-Zn and Co-Zn heteronuclear carbonyl cation complexes are produced via a laser vaporization supersonic cluster source in the gas phase. The dinuclear FeZn(CO)5+ and CoZn(CO)7+ cation complexes are observed to be the most intense heterodinuclear carbonyl cation species in the mass spectra. The infrared spectra are obtained via mass selection and infrared photodissociation spectroscopy in the carbonyl stretching frequency region. Their geometric and electronic structures are assigned with the support of density functional calculations. The FeZn(CO)5+ complex is determined to have a (OC)5Fe-Zn structure with a Fe-Zn half bond. The CoZn(CO)7+ ion is established to have a staggered (OC)4Co-Zn(CO)3 structure involving a Co-Zn σ single bond.

Infrared Photodissociation Spectroscopic and Theoretical Study of Heteronuclear Transition Metal Carbonyl Cluster Cations in the Gas Phase.

Heteronuclear transition metal carbonyl cluster cations FeM(CO)8(+) (M = Co, Ni and Cu) and MCu(CO)7(+) (M = Co and Ni) are produced via a laser vaporization supersonic cluster ion source in the gas phase, which are each mass-selected and studied by infrared photodissociation spectroscopy in the carbonyl stretching frequency region. Their geometric and electronic structures are established by comparison of the experimental spectra with those derived from density functional theoretical calculations. The FeM(CO)8(+) (M = Co, Ni, Cu) complexes are determined to have eclipsed (CO)5Fe-M(CO)3(+) structures, and the MCu(CO)7(+) (M = Co, Ni) ions are characterized to have staggered (CO)4M-Cu(CO)3(+) structures. Natural bonding orbital analysis indicates that the positive charge is mainly distributed on the M(CO)3 fragment. The metal-metal interaction involves an σ-type (OC)4,5M→M(CO)3(+) dative bonding.