Unveiling the potential of Butylphthalide: inhibiting osteoclastogenesis and preventing bone loss

Osteoporosis, resulting from overactive osteoclasts and leading to elevated fracture risk, has emerged as a global public health concern due to the aging population. Therefore, inhibiting osteoclastogenesis and bone resorption function represents a crucial approach for preventing and treating osteoporosis. The purpose of this study was to examine the effects and molecular mechanisms of Butylphthalide (NBP) on the differentiation and function of osteoclasts induced by RANKL. Osteoclastogenesis was assessed through TRAP staining and bone slice assay. An animal model that underwent ovariectomy, simulating postmenopausal women’s physiological characteristics, was established to investigate the impact of Butylphthalide on ovariectomy-induced bone loss. To delve deeper into the specific mechanisms, we employed Western blot, PCR, immunofluorescence, and immunohistochemical staining to detect the expression of proteins that are associated with the osteoclast signaling pathway. In this study, we found that Butylphthalide not only suppressed osteoclastogenesis and bone resorption in vitro but also significantly decreased TRAcP-positive osteoclasts and prevented bone loss in vivo. Further mechanistic experiments revealed that Butylphthalide reduces intracellular ROS in osteoclasts, inhibits the MAPK and NFATc1 signaling pathways, and downregulates the key genes and proteins of osteoclasts. This inhibits osteoclast formation and function. The reduction in ROS in osteoclasts is intricately linked to the activity of Butylphthalide-modulated antioxidant enzymes. Overall, NBP may offer a alternative treatment option with fewer side effects for skeletal diseases such as osteoporosis

miR-136-5p Regulates the Inflammatory Response by Targeting the IKKβ/NF-κB/A20 Pathway After Spinal Cord Injury

Background/Aims: miR-136-5p participates in recovery after spinal cord injury (SCI) via an unknown mechanism. We investigated the mechanism underlying the involvement of miR-136-5p in the inflammatory response in a rat model of SCI. Methods: Sprague-Dawley rat astrocytes were cultured in vitro to construct a reporter plasmid. Luciferase assays were used to detect the ability of miR-136-5p to target the IKKβ and A20 genes. Next, recombinant lentiviral vectors were constructed, which either overexpressed miR-136-5p or inhibited its expression. The influence of miR-136-5p overexpression and miR-136-5p silencing on inflammation was observed in vivo in an SCI rat model. The expression of IL-1β, IL-6, TNF-α, IFN-α, and related proteins (A20, IKKβ, and NF-κB) was detected. Results: In vitro studies showed that luciferase activity was significantly activated in the presence of the 3’ untranslated region (UTR) region of the IKKβ gene after stimulation of cells with miR-136-5p. However, luciferase activity was significantly inhibited in the presence of the 3’UTR region of the A20 gene. Thus, miR-136-5p may act directly on the 3’UTR regions of the IKKβ and A20 genes to regulate their expression. miR-136-5p overexpression promoted the production of related cytokines and NF-κB in SCI rats and inhibited the expression of A20 protein. Conclusion: Overexpression of miR-136-5p promotes the generation of IL-1β, IL-6, TNF-α, IFN-α, IKKβ, and NF-κB in SCI rats but inhibits the expression of A20. Under these conditions, inflammatory cell infiltration into the rat spinal cord increases and injury is significantly aggravated. Silencing of miR-136-5p significantly reduces the protein expression results described after miR-136-5p overexpression and ameliorates the inflammatory cell infiltration and damage to the spinal cord. Therefore, miR-136-5p might be a new target for the treatment of SCI

Treatment results in the differential surgery of intradural extramedullary schwannoma of 110 cases.

STUDY DESIGN: A retrospective study of intradural extramedullary schwannoma. OBJECTIVE: The purpose of this study was to compare treatment results in the differential surgery of intradural extramedullary schwannoma. BACKGROUND: A reference guide to the surgical procedures available to treat intradural extramedullary schwannoma has not yet been established. METHODS: The study retrospectively reviewed 110 patients: Group A: laminectomy+microscopic excision; Group B: hemilaminectomy+microscopic excision; Group C: laminectomy+microscopic excision+pedicle screw fixation. Researchers selected patients for this retrospective review by applying the following criteria: 1) back pain spread out from the tumor level, sensory and motor loss; 2) treatment by surgery; 3) clinical diagnosis made by physical examination, magnetic resonance imaging (MRI), and pathology; 4) a minimum clinical and radiologic follow-up of 12 months. The clinical outcomes were assessed by comparing the Visual Analogue Pain Scores (VAS) and the Japanese Orthopedic Association Scores (JOA score). The study also performed a cost-effectiveness analysis. RESULTS: Cervical vertebrae: The estimated blood loss in Group B was significantly less than in Group C (P<0.05) (Table 1). Thoracic vertebrae: The duration of hospital stay and estimated blood loss in Group A was significantly less than in Group C (P<0.05) (Table 2, 3). Lumbar vertebrae: The resection rate in Group C was significantly higher than in Group A and Group B (P<0.05) (Table 4). Treatment in Group B was the least expensive, and therefore, the most cost-effective. CONCLUSION: In the case of appropriate surgical indications, the study suggests that hemilaminectomy+microscopic excision is advantageous in the removal of cervical schwannoma, and that laminectomy+microscopic excision is advantageous in the removal of thoracic schwannoma; lumbar intradural extramedullary schwannoma can be managed by laminectomy+microscopic excision+pedicle screw fixation

The Effects of Interleukin-17 (IL-17)-Related Inflammatory Cytokines and A20 Regulatory Proteins on Astrocytes in Spinal Cord Cultured In Vitro

Background/Aims: This study focused on investigating the expression of several inflammatory cytokines and chemokines, including regulatory proteins in the astrocytes of mice stimulated with IL-17. Materials and Methods: The cultured astrocytes from the spinal cords of mice were stimulated with IL-17. The expression of interleukin-6 (IL-6), tumor necrosis factor (TNF), monocyte chemotactic protein-1/5 (MCP-1/5) and macrophage inflammatory protein-2 (MIP-2) stimulated with IL-17 (50 ng/ml) at different time points (3 h, 6 h, 12 h, 24 h and 48 h) were determined using real-time PCR and ELISA. The expressions of A20 (tumor necrosis factor a inducible protein 3, TNFAIP3) and NF-&#954;B were examined using real-time PCR and western blotting. Results: Compared with the control group, the mRNA expression levels of IL-6, TNF, MCP-1/5 and MIP-2 increased significantly at 6 h after IL-17 stimulation, while the protein expression levels also increased significantly and peaked at 12 h. The mRNA expression level of NF-&#954;B increased and peaked at 6 h before gradually declining, while the expression of A20 decreased. The protein expression level of NF-&#954;B increased and peaked at 12 h, while the expression A20 had an opposite response. Conclusion: The study showed that NF-&#954;B may have an effect on the cytokines secreted by astrocytes after IL-17 stimulation. Moreover, both A20 and NF-&#954;B could regulate the expression and secretion of inflammatory mediators

Molecular Mechanism of MiR-136-5p Targeting NF-κB/A20 in the IL-17-Mediated Inflammatory Response after Spinal Cord Injury

Background/Aims: The pathophysiology of spinal cord injury (SCI) results in serious damage to the human body via an increase in the secondary biological processes imposed by activated astrocytes. Abnormal expression of microRNAs after SCI has become a potential research focus. However, the underlying mechanisms are poorly understood. Methods: SCI models were established in rats using Allen’s method, and the BBB scoring method was employed to assess locomotor function. Lentivirus was used to infect rat astrocytes and SCI rats. Real-time PCR and antibody chip were used to measure gene expression and cytokine secretion. Western blot analysis was employed to detect protein expression. HE staining was used to assess the histological changes in SCI. The immunohistochemical staining of A20 and p-NF-κB in SCI was also analyzed. Results: The in vitro experiment showed that miR-136-5p up-regulated the expression of p-NF-κB by down-regulating the expression of A20 so that astrocytes produced inflammatory factors and chemokines. The in vivo experiment indicated that overexpressed miR-136-5p promoted the production of inflammatory factors, chemokines and p-NF-κB in SCI rats, whereas it inhibited the expression of A20 protein and increased inflammatory cell infiltration and injuries in the spinal cord. Conclusion: The current findings indicate that silencing miR-136-5p effectively decreased inflammatory factors and chemokines and protected the spinal cord via NF-κB/A20 signaling in vivo and in vitro. In contrast, overexpression of miR-136-5p had the opposite effect

Clinical outcomes of the levels of schwannoma at thoracic vertebrae.

<p>This table shows that the clinical outcomes of the levels of schwannoma at thoracic vertebrae.</p><p>Data are expressed as the mean ± standard deviation.</p

Clinical outcomes of the levels of schwannoma at lumbar vertebrae.

<p>This table shows that the clinical outcomes of the levels of schwannoma at lumbar vertebrae.</p><p>PRE: preoperative; POST: postoperative; JOA-C: Japanese Orthopedic Association scores (JOA scores) for cervical vertebra; VAS: Visual Analogue Pain Score.</p><p>Data are expressed as the mean ± standard deviation.</p

Summary of baseline characteristics of intradural extramedullary schwannoma patients.

<p>This table showed that there was no significant difference among the three groups (Group A, Group B, Group C) in the basic characteristics (Age distribution, Gender distribution, Course of disease, and Tumor size).</p><p>CV: Cervical vertebra; TV: Thoracic vertebra; LV: Lumbar vertebra.</p><p>Data are expressed as the mean ± standard deviation.</p