Anatomical Analysis of the Cervical Sympathetic Ganglion and Spinal Ganglion and its Physiological Significance in the Pathogenesis of Cervical Vertigo.

Objective: Investigating the anatomical connections between cervical sympathetic ganglia and spinal ganglia in rabbits and assessing the role of Neuropeptide Y in the pathogenesis of cervical vertigo. Method: Part 1: 32 adult healthy male New Zealand white rabbits (whose skin is very sensitive, so rabbits are generally used for stimulation experiments) were randomly divided into the upper cervical sympathetic ganglia (SCSG) stimulation group and the lower cervical sympathetic ganglia (ICSG) stimulation group, with 16 rabbits in each group. The two groups were divided into an experimental group and a control group, with 8 rabbits in each group. The cervical ganglia of each group of white rabbits were injected with 4% FluoroGold solution and observed under a section microscope. Part 2: Sixty New Zealand white rabbits were randomly divided into a blank control (n = 12), SCSG stimulation group (n = 12), SCSG sham surgery control (n = 12), ICSG stimulation group (n = 12), and ICSG sham surgery control group (n = 12). The SCSG group and ICSG group were subjected to electrical stimulation (i.e. 30.0Hz, 10.0V, 5-minute pulse width of 0.5 ms square wave pulse), and specimens were made. The expression of NPY was detected using immunohistochemical methods. Result: Neuropeptide Y was weakly expressed in all cervical ganglia (C1-C8). Compared with the sham surgery group, the superior cervical sympathetic ganglion stimulation group showed an increase in Neuropeptide Y positive cells in C2, C3, C4, and C5, with C2 and C3 showing the most significant increase. The number of C6, C7, and C8 Neuropeptide Y positive cells in the 3 C、3D and 4B, lower cervical sympathetic ganglion stimulated groups was higher than in the sham sham-operated group, and C6 and C7 significantly increased. Neuropeptide Y is like immunoreactive neurons in the cervical spinal ganglia, and the immunoreactive products are small brown particles distributed in the cytoplasm after electrical stimulation of the cervical sympathetic ganglia. The Neuropeptide Y content in the corresponding segment of the cervical spinal ganglia is significantly increased compared to the control group (P < .05). Conclusion: In New Zealand white rabbits, nerve fibers are interconnected between the cervical sympathetic ganglion and the cervical spinal ganglion, and this neural fiber connection has a certain segmental nature, providing experimental basis for the existence of the cervical spinal cord external nerve reflex arc and elucidating the pathogenesis of cervical vertigo in terms of neural anatomy. By using neuroelectrophysiological methods, it has been confirmed that electrical stimulation in the cervical spinal ganglia can reach the corresponding cervical sympathetic ganglia on the same side through a certain conduction pathway, providing experimental basis in neuroelectrophysiology for the existence of the cervical extraspinal nerve reflex arc and elucidating the pathogenesis of cervical vertigo. NPY may be involved in the pathogenesis of cervical vertigo, providing a theoretical basis for the clinical diagnosis of cervical vertigo.

Regulator of G protein signalling 18 promotes osteocyte proliferation by activating the extracellular signal­regulated kinase signalling pathway.

Osteocyte function is critical for metabolism, remodelling and regeneration of bone tissue. In the present study, the roles of regulator of G protein signalling 18 (RGS18) were assessed in the regulation of osteocyte proliferation and bone formation. Target genes and signalling pathways were screened using the Gene Expression Omnibus (GEO) database and Gene Set Enrichment Analysis (GSEA). The function of RGS18 and the associated mechanisms were analysed by Cell Counting Kit 8 assay, 5­ethynyl­2'­deoxyuridine assay, flow cytometry, reverse transcription­quantitative PCR, western blotting and immunostaining. Overlap analysis of acutely injured subjects (AIS) and healthy volunteers (HVs) from the GSE93138 and GSE93215 datasets of the GEO database identified four genes: KIAA0825, ANXA3, RGS18 and LIPN. Notably, RGS18 was more highly expressed in peripheral blood samples from AIS than in those from HVs. Furthermore, RGS18 overexpression promoted MLO­Y4 and MC3T3­E1 cell viability, proliferation and S­phase arrest, but inhibited apoptosis by suppressing caspase­3/9 cleavage. Silencing RGS18 exerted the opposite effects. GSEA of GSE93138 revealed that RGS18 has the ability to regulate MAPK signalling. Treatment with the MEK1/2 inhibitor PD98059 reversed the RGS18 overexpression­induced osteocyte proliferation, and treatment with the ERK1/2 activator 12­O­tetradecanoylphorbol­13­acetate reversed the effects of RGS18 silencing on osteocyte proliferation. In conclusion, RGS18 may contribute to osteocyte proliferation and bone fracture healing via activation of ERK signalling.

Abrine promotes cell proliferation and inhibits apoptosis of interleukin-1ß-stimulated chondrocytes via PIM2/VEGF signalling in osteoarthritis.

BACKGROUND: Osteoarthritis (OA), a common joint disorder with an increasing incidence worldwide, severely affects the quality of life of patients. In Chinese herbal medicine, Abrus cantoniensis Hance is considered to exert protective effects on the liver and to have beneficial effects on the gallbladder; additionally, it has antibacterial and anti-inflammatory properties, as well as the ability to enhance immunity, scavenge free radicals, regulate smooth muscle function, and improve endurance. Abrine extracted from A. cantoniensis Hance has been reported as a main functional compound capable of treating chronic inflammation. PURPOSE: In this study, we explored the effect of abrine on OA progression. STUDY DESIGN: Bioinformatics analysis was performed on abrine and its potential targets in OA, using the Comparative Toxicogenomics Database, GSE1919 dataset from the Gene Expression Omnibus database, Gene Set Enrichment Analysis, and docking interaction analysis. METHODS: The effect of abrine in vitro was analysed by Cell Counting Kit 8 assays, colony formation assays, enzyme-linked immunosorbent assay, flow cytometry analysis, quantitative real-time PCR, and western blotting using human transformed chondrocyte cell line C28/I2. The effect of abrine was evaluated in vivo using the anterior cruciate ligament transection (ACLT) Sprague-Dawley rat OA model. RESULTS: Abrine enhanced the proliferation of interleukin (IL)-1ß-stimulated C28/I2 cells in a dose-dependant manner. Expression of pro-inflammatory cytokines was induced by IL-1ß treatment, whereas abrine treatment repressed the induction of C28/I2 cells in a dose dependant manner (p < 0.05). Abrine induced cell proliferation and inhibited apoptosis in IL-1ß-stimulated C28/I2 cells (p < 0.05). Abrine also inhibited Proviral Integrations of Moloney virus 2 (PIM2) expression in IL-1ß-stimulated C28/I2 cells (p < 0.05). The expression of vascular endothelial growth factor (VEGF), p-VEGFR2, and p-eNOS was induced by IL-1ß treatment in C28/I2 cells, while abrine inhibited this induction in a dose dependant manner. Treatment with abrine decreased the expression levels of PIM2 and VEGF in IL-1ß-stimulated C28/I2 cells (p < 0.05). Overexpression of PIM2 induced cell proliferation and inhibited apoptosis in IL-1ß-stimulated C28/I2 cells, while VEGF silencing reversed this effect (p < 0.05). Finally, abrine prevented cartilage degradation in the ACLT model. CONCLUSION: We demonstrated that abrine promoted cell proliferation and inhibited apoptosis in IL-1ß-stimulated C28/I2 cells through PIM2/VEGF signalling. These findings indicate PIM2 to be a potential drug target. Moreover, abrine has potential applicability as a therapeutic agent against OA.

Knockdown of exosome­mediated lnc­PVT1 alleviates lipopolysaccharide­induced osteoarthritis progression by mediating the HMGB1/TLR4/NF­κB pathway via miR­93­5p.

Osteoarthritis is a chronic degenerative joint disease. Long non­coding RNA plasmacytoma variant translocation 1 (PVT1) is involved in the progression of osteoarthritis and exosomes serve a central role in intercellular communication. However, whether PVT1 can be mediated by exosomes in osteoarthritis has not been reported. Whole blood was drawn from osteoarthritis patients and healthy volunteers. Lipopolysaccharide (LPS) was used to stimulate human normal chondrocytes (C28/I2) to construct a cell damage model in vitro. Protein levels were examined via western blot analysis. eThe expression of PVT1, microRNA (miR)­93­5p and high mobility groupprotein B1 (HMGB1) was evaluated through reverse transcription­quantitative PCR. Cell viability and apoptosis were determined through CCK­8 or flow cytometric assay. Inflammatory cytokines were measured via ELISA. The relationship between PVT1 or HMGB1 and miR­93­5p was confirmed by dual­luciferase reporter assay. PVT1, HMGB1 and exosomal PVT1 were upregulated while miR­93­5p was downregulated in osteoarthritis patient serum and LPS­induced C28/I2 cells. Exosomes from osteoarthritis patient serum and LPS­treated C28/I2 cells increased PVT1 expression in C28/I2 cells. PVT1 depletion reversed the decrease of viability and the increase of apoptosis, inflammation responses and collagen degradation of C28/I2 cells induced by LPS. PVT1 regulated HMGB1 expression via sponging miR­93­5p. miR­93­5p inhibition abolished PVT1 silencing­mediated viability, apoptosis, inflammation responses and collagen degradation of LPS­stimulated C28/I2 cells. HMGB1 increase overturned miR­93­5p upregulation­mediated viability, apoptosis, inflammation responses and collagen degradation of LPS­stimulated C28/I2 cells. Furthermore, PVT1 modulated the Toll­like receptor 4/NF­κB pathway through an miR­93­5p/HMGB1 axis. In summary, exosome­mediated PVT1 regulated LPS­induced osteoarthritis progression by modulating the HMGB1/TLR4/NF­κB pathway via miR­93­5p, providing a new route for possible osteoarthritis treatment.

Functional Pathway Between Cervical Spinal and Sympathetic Ganglia: A Neurochemical Foundation Between Neck Pain and Vertigo.

BACKGROUND: Cervical vertigo commonly concurs in patients with neck pain, but the concurrent mechanism of these 2 symptoms still remains unclear. We previously reported a bidirectional segmental nerve fiber connection between cervical spinal and sympathetic ganglia, which provided a hypothesis that this connection between the 2 ganglia may be the anatomic basis for the concurrence of neck pain and cervical vertigo. However, this concurrent mechanism needs biochemical and functional evidence. OBJECTIVES: This study aimed to investigate a possible noradrenergic pathway between cervical spinal and sympathetic ganglia. STUDY DESIGN: We performed both clinical and laboratory research. Clinical observation was a prospective case-control study. SETTING: Clinical study took place in our hospital; laboratory study was in an orthopedic laboratory. METHODS: Cervical lamina block therapy used in patients with cervical vertigo was clinically evaluated; norepinephrine (NE) expressions in cervical sympathetic ganglia were analyzed using immunohistochemical staining after electrical stimulation to the cervical spinal ganglia; the influence of phentolamine local injection to the vertebrobasilar artery flow was experimentally measured. RESULTS: Cervical lamina block therapy could significantly shorten the clinical hospital stays of patients with cervical vertigo (P = 0.000) and improve vertebral artery flow (P < 0.05). NE expressions in superior cervical sympathetic ganglia (SCG) or inferior cervical sympathetic ganglia (ICG) increased significantly when ipsilateral C2 to C3 or C6 to C8 spinal ganglia were electrically stimulated, respectively. Adrenergic receptor block with phentolamine significantly inhibited the decrease of basilar artery (BA) flow induced by electrical stimulation of the cervical spinal ganglia. The change range of BA flow caused by stimulations of C2 to C3 and C6 to C8 spinal ganglia was more than that of C4 and C5. LIMITATIONS: The inpatients observed in this clinical study might be influenced by some factors including emotion, diet, sleep, and others. The limitations of the laboratory study included animal species and small sample size. CONCLUSIONS: Adrenergic system could play a part in cervical spinal ganglia altering the vertebrobasilar artery system. It could provide a neurochemical foundation between neck pain and vertigo, and that segmental functional connections exist between cervical spinal and sympathetic ganglia. KEY WORDS: Cervical vertigo, neck pain, cervical sympathetic ganglia, cervical spinal ganglia, noradrenaline.

Neural reflex pathway between cervical spinal and sympathetic ganglia in rabbits: implication for pathogenesis of cervical vertigo.

BACKGROUND CONTEXT: A functional association between cervix and vertigo has been observed in patients with cervical vertigo, implicating correlation between cervical spinal and sympathetic ganglia. However, it is unclear where there is an anatomic connection between those two groups of ganglia. PURPOSE: This study aimed to investigate the existence of the neural connections between cervical spinal and sympathetic ganglia. STUDY DESIGN/SETTING: FluoroGold staining patterns in cervical spinal and sympathetic ganglia were evaluated using FluoroGold retrograde tracing in New Zealand rabbits. METHODS: New Zealand rabbits were randomly divided into superior cervical spinal ganglion injection groups, inferior cervical spinal ganglion injection groups, superior cervical sympathetic ganglion injection group, and inferior cervical sympathetic ganglion injection group. Four percent FluoroGold solution was injected into these ganglia. Distribution of FluoroGold in cervical spinal and sympathetic ganglia was observed under a microscope. RESULTS: When FluoroGold solution was injected into C2 and C3 superior cervical spinal ganglia or C5-C6 inferior cervical spinal ganglia, fluorescence was only observed in the ipsilateral superior or inferior cervical sympathetic ganglia, respectively. When FluoroGold solution was injected into superior or inferior cervical sympathetic ganglia, fluorescence was found mainly in the ipsilateral C3-C4 superior or C5-C8 inferior spinal ganglia. No fluorescence was observed in contralateral ganglia of experimental animals and all ganglia of matched control animals injected with physiological saline. CONCLUSIONS: Bidirectional nerve fiber connections between cervical spinal and sympathetic ganglia were observed, and these connections are arranged in a segmental distribution. This observation may provide a possible neuroanatomic basis for the pathogenesis of cervical vertigo.