RESUMO
Notch signal plays an important role in regulating cell-cell interactions with the adjacent cells. However, it remains unknown whether Jagged1 (JAG-1) mediated Notch signaling regulates bone cancer pain (BCP) via the spinal cell interactions mechanism. Here, we showed that intramedullary injection of Walker 256 breast cancer cells increased the expression of JAG-1 in spinal astrocytes and knockdown of JAG-1 reduced BCP. The supplementation of exogenous JAG-1 to the spinal cord induced BCP-like behavior and promoted expression of c-Fos and hairy and enhancer of split homolog-1 (Hes-1) in the spinal cord of the naïve rats. These effects were reversed when the rats were administered intrathecal injections of N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester (DAPT). The intrathecal injection of DAPT reduced BCP and inhibited Hes-1 and c-Fos expression in the spinal cord. Furthermore, our results showed that JAG-1 up-regulated Hes-1 expression by inducing the recruitment of Notch intracellular domain (NICD) to the RBP-J/CSL-binding site located within the Hes-1 promoter sequence. Finally, the intrathecal injection of c-Fos-antisense oligonucleotides (c-Fos-ASO) and administration of sh-Hes-1 to the spinal dorsal horn also alleviated BCP. The study indicates that inhibition of the JAG-1/Notch signaling axis may be a potential strategy for the treatment of BCP.
Assuntos
Neoplasias Ósseas , Dor do Câncer , Ratos , Animais , Dor do Câncer/etiologia , Neoplasias Ósseas/complicações , Transdução de Sinais/fisiologia , Dor , Medula EspinalRESUMO
Due to its complex pathological mechanisms, bone cancer pain (BCP) has become an increasingly challenging clinical issue, there is an urgent need to identify the underlying mechanisms of BCP. In our present study, we found that decreased expression of miR-199a-3p in spinal dorsal horn (SDH) neurons contributed to BCP hypersensitivity. Intrathecal administration of miR-199a-3p agomir alleviated the initiation of tumor inoculation induced pain hypersensitivity and suppressed the expression of DNMT3A. Subsequently, luciferase assays confirmed direct binding between miR-199a-3p and Dnmt3a mRNA. AAV-DNMT3A-shRNA microinjection relieved mechanical hyperalgesia and upregulated the expression of Nrf2 levels in BCP. In naïve rats, Overexpression of DNMT3A yielded the opposite effects. Finally, increase of DNMT3A by lentiviral vector abolished miR-199a-3p-mediated alleviation hypersensitivity effects on BCP progression. Taken these together, our findings highlighted a novel contribution of miR-199a-3p to BCP and provided a fresh outlook on potential mechanism research for BCP.
Assuntos
Neoplasias Ósseas , Dor do Câncer , MicroRNAs , Osteossarcoma , Ratos , Animais , Dor do Câncer/genética , Dor do Câncer/metabolismo , Regulação para Cima , Dor/metabolismo , Neoplasias Ósseas/complicações , Neoplasias Ósseas/genética , Neoplasias Ósseas/metabolismo , Células do Corno Posterior/metabolismo , Osteossarcoma/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismoRESUMO
BACKGROUND: Complex regional pain syndrome type-I (CRPS-I) causes excruciating pain that affect patients' life quality. However, the mechanisms underlying CRPS-I are incompletely understood, which hampers the development of target specific therapeutics. METHODS: The mouse chronic post-ischemic pain (CPIP) model was established to mimic CRPS-I. qPCR, Western blot, immunostaining, behavioral assay and pharmacological methods were used to study mechanisms underlying neuroinflammation and chronic pain in spinal cord dorsal horn (SCDH) of CPIP mice. RESULTS: CPIP mice developed robust and long-lasting mechanical allodynia in bilateral hindpaws. The expression of inflammatory chemokine CXCL13 and its receptor CXCR5 was significantly upregulated in ipsilateral SCDH of CPIP mice. Immunostaining revealed CXCL13 and CXCR5 was predominantly expressed in spinal neurons. Neutralization of spinal CXCL13 or genetic deletion of Cxcr5 (Cxcr5-/-) significantly reduced mechanical allodynia, as well as spinal glial cell overactivation and c-Fos activation in SCDH of CPIP mice. Mechanical pain causes affective disorder in CPIP mice, which was attenuated in Cxcr5-/- mice. Phosphorylated STAT3 co-expressed with CXCL13 in SCDH neurons and contributed to CXCL13 upregulation and mechanical allodynia in CPIP mice. CXCR5 coupled with NF-κB signaling in SCDH neurons to trigger pro-inflammatory cytokine gene Il6 upregulation, contributing to mechanical allodynia. Intrathecal CXCL13 injection produced mechanical allodynia via CXCR5-dependent NF-κB activation. Specific overexpression of CXCL13 in SCDH neurons is sufficient to induce persistent mechanical allodynia in naïve mice. CONCLUSIONS: These results demonstrated a previously unidentified role of CXCL13/CXCR5 signaling in mediating spinal neuroinflammation and mechanical pain in an animal model of CRPS-I. Our work suggests that targeting CXCL13/CXCR5 pathway may lead to novel therapeutic approaches for CRPS-I.
Assuntos
Quimiocina CXCL13 , Dor Crônica , Receptores CXCR5 , Distrofia Simpática Reflexa , Animais , Camundongos , Quimiocina CXCL13/metabolismo , Modelos Animais de Doenças , Hiperalgesia , Doenças Neuroinflamatórias , NF-kappa B , Corno Dorsal da Medula Espinal , Receptores CXCR5/metabolismoRESUMO
Bone cancer pain (BCP) is a clinically intractable mixed pain, involving inflammation and neuropathic pain, and its mechanisms remain unclear. CXC chemokine receptor 1 (CXCR1, IL-8RA) and 2 (CXCR2, IL-8RB) are high-affinity receptors for interleukin 8 (IL8). According to previous studies, CXCR2 plays a crucial role in BCP between astrocytes and neurons, while the role of CXCR1 remains unclear. The objective of this study was to investigate the role of CXCR1 in BCP. We found that CXCR1 expression increased in the spinal dorsal horn. Intrathecal injection of CXCR1 siRNA effectively attenuated mechanical allodynia and pain-related behaviors in rats. It was found that CXCR1 was predominantly co-localized with neurons. Intrathecal injection of CXCR1-siRNA reduced phosphorylated JAK2/STAT3 protein levels and the NLRP3 inflammasome (NLRP3, caspase1, and IL-1ß) levels. Furthermore, in vitro cytological experiments confirmed this conclusion. The study results suggest that the spinal chemokine receptor CXCR1 activation mediates BCP through JAK2/STAT3 signaling pathway and NLRP3 inflammasome (NLRP3, caspase1, and IL-1ß).
Assuntos
Neoplasias Ósseas , Dor do Câncer , Neuralgia , Ratos , Feminino , Animais , Receptores de Interleucina-8A/genética , Receptores de Interleucina-8A/metabolismo , Dor do Câncer/etiologia , Dor do Câncer/metabolismo , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , RNA Interferente Pequeno/metabolismo , Neoplasias Ósseas/complicações , Neoplasias Ósseas/metabolismo , Receptores de Interleucina-8B/metabolismo , Neuralgia/metabolismo , Medula Espinal/metabolismoRESUMO
BACKGROUND: Noncoding microRNAs have emerged as critical players of gene expression in the nervous system, where they contribute to regulating nervous disease. As stated in previous research, the miR-155-5p upregulation happens in the spinal cord at the nociceptive state. It was unclear if miR-155-5p is linked to bone cancer pain (BCP). Herein, we aimed at investigating the miR-155-5p functional regulatory function in BCP process and delineating the underlying mechanism. METHODS: The miRNA-155-5p levels and cellular distribution were determined by RNA sequencing, fluorescent in situ hybridization (FISH), and quantitative real-time PCR (qPCR). Immunoblotting, qPCR, dual-luciferase reporter gene assays, immunofluorescence, recombinant overexpression adeno-associated virus, small interfering RNA, intraspinal administration, and behavioral tests were utilized for exploring the downstream signaling pathway. RESULTS: The miR-155-5p high expression in spinal neurons contributes to BCP maintenance. The miR-155-5p blockage via the intrathecal injection of miR-155-5p antagomir alleviated the pain behavior; in contrast, upregulating miR-155-5p by agomir induced pain hypersensitivity. The miR-155-5p bounds directly to TCF4 mRNA's 3' UTR. BCP significantly reduced protein expression of TCF4 versus the Sham group. The miR-155-5p inhibition relieved the spinal TCF4 protein's down-expression level, while miR-155-5p upregulation by miR-155-5p agomir intrathecal injection decreased TCF4 protein expression in naïve rats. Additionally, TCF4 overexpression in BCP rats could increase Kv1.1. Moreover, TCF4 knockdown inhibited Kv1.1 expression in BCP rats. Indeed, TCF4 and Kv1.1 were co-expressed in BCP spinal cord neurons. CONCLUSION: The study findings stated the miR-155-5p pivotal role in regulating BCP by directly targeting TCF4 in spinal neurons and suggested that miR-155-5p could be a promising target in treating BCP.
Assuntos
Neoplasias Ósseas , Dor do Câncer , MicroRNAs , Regiões 3' não Traduzidas , Animais , Antagomirs/metabolismo , Neoplasias Ósseas/complicações , Neoplasias Ósseas/genética , Neoplasias Ósseas/metabolismo , Dor do Câncer/genética , Dor do Câncer/metabolismo , Hibridização in Situ Fluorescente , Luciferases/genética , Luciferases/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Dor/metabolismo , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/metabolismo , Ratos , Medula Espinal/metabolismoRESUMO
Bone cancer pain (BCP) is a clinical pathology that urgently needs to be solved, but research on the mechanism of BCP has so far achieved limited success. Nuclear factor erythroid 2 (NFE2)-related factor 2 (Nrf2) has been shown to be involved in pain, but its involvement in BCP and the specific mechanism have yet to be examined. This study aimed to test the hypothesis that BCP induces the transfer of Nrf2 from the cytoplasm to the nucleus and further promotes nuclear transcription to activate heme oxygenase-1 (HO-1) and inhibit the activation of nuclear factor-kappa B (NF-κB) signalling, ultimately regulating the neuroinflammatory response. Von-Frey was used for behavioural analysis in rats with BCP, whereas western blotting, real-time quantitative PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect molecular expression changes, and immunofluorescence was used to detect cellular localization. We demonstrated that BCP induced increased Nrf2 nuclear protein expression with decreased cytoplasmic protein expression in the spinal cord. Further increases in Nrf2 nuclear protein expression can alleviate hyperalgesia and activate HO-1 to inhibit the expression of NF-κB nuclear protein and inflammatory factors. Strikingly, intrathecal administration of the corresponding siRNA reversed the above effects. In addition, the results of double immune labelling revealed that Nrf2 and NF-κB were coexpressed in spinal cord neurons of rats with BCP. In summary, these findings suggest that the entry of Nrf2 into the nucleus promotes the expression of HO-1, inhibiting activation of the NF-κB signalling pathway, reducing neuroinflammation and ultimately exerting an anti-nociceptive effect.
Assuntos
Neoplasias Ósseas/metabolismo , Dor do Câncer/metabolismo , Hiperalgesia/metabolismo , Fator 2 Relacionado a NF-E2/biossíntese , NF-kappa B/metabolismo , Medula Espinal/metabolismo , Transporte Ativo do Núcleo Celular/fisiologia , Animais , Neoplasias Ósseas/patologia , Dor do Câncer/patologia , Linhagem Celular Tumoral , Núcleo Celular/metabolismo , Feminino , Hiperalgesia/patologia , NF-kappa B/antagonistas & inibidores , Neurônios/metabolismo , Neurônios/patologia , Ratos , Ratos Sprague-Dawley , Medula Espinal/patologiaRESUMO
BACKGROUND Primary lower-extremity hyperhidrosis (PLEH) can be treated by CT-guided lumbar sympathetic nerve modulation using absolute ethanol. However, doses of ethanol that are too high can cause nerve injury, and doses that are too low have suboptimal results. The present study aimed to investigate the dose-effect relationship of CT-guided lumbar sympathetic nerve modulation with absolute ethanol for PLEH. MATERIAL AND METHODS The study was conducted at the First Affiliated Hospital of Jiaxing University between 07/2014 and 02/2017. Twenty participants were enrolled in each group. The doses of absolute ethanol were 2.0 ml in the R1 group, 2.5 ml in the R2 group, 3.0 ml in the R3 group, 3.5 ml in the R4 group, and 4.0 ml in the R5 group. Treatment effectiveness was assessed according to the time to complete hyperhidrosis relief: <10 min, effective; ≥10 min, non-effective. RESULTS The patient characteristics among the 5 groups were not statistically different (P>0.05). The onset time and time to complete hyperhidrosis relief decreased significantly with increasing dose of absolute ethanol (P<0.05). The effective rates in the 5 groups were 15.0%, 35.0%, 60.0%, 90.0%, and 100.0%, respectively. The ED50 and ED95 were 2.306 ml (95% CI: 2.003-2.512 ml) and 3.343 ml (95% CI: 3.051-3.962 ml), respectively. CONCLUSIONS This was the first dose-effect pilot study of consecutive PLEH patients treated by CT-guided lumbar sympathetic nerve modulation. CT-guided lumbar sympathetic nerve modulation with 2.306 ml (ED50) and 3.343 ml (ED95) of absolute ethanol showed treatment efficacy for PLEH. No complications were seen.
Assuntos
Bloqueio Nervoso Autônomo/métodos , Etanol/farmacologia , Hiperidrose/terapia , Extremidade Inferior/fisiopatologia , Adulto , Relação Dose-Resposta a Droga , Etanol/administração & dosagem , Etanol/uso terapêutico , Feminino , Humanos , Extremidade Inferior/inervação , Plexo Lombossacral/diagnóstico por imagem , Plexo Lombossacral/efeitos dos fármacos , Masculino , Tomografia Computadorizada por Raios X/métodosRESUMO
Chronic pain is one of the serious conditions that affect human health and remains cure still remains a serious challenge as the molecular mechanism remains largely unclear. Here, we used label-free proteomics to identify potential target proteins that regulate peripheral inflammatory pain and reveal its mechanism of action. Inflammation in peripheral tissue was induced by injecting complete Freund's adjuvant (CFA) into rat hind paw. A proteomic method was adopted to compare the spinal dorsal horn (SDH) in peripheral inflammatory pain (PIP) model rats with controls. Differential proteins were identified in SDH proteome by label-free quantification. The role of screened target proteins in the PIP was verified by small interfering RNA (siRNA). A total of 3072 and 3049 proteins were identified in CFA and normal saline (NS) groups, respectively, and 13 proteins were identified as differentially expressed in the CFA group. One of them, neurexin-2, was validated for its role in the inflammatory pain. Neurexin-2 was up-regulated in the CFA group, which was confirmed by quantitative PCR. Besides, intrathecal siRNA-mediated knock-down of neurexin-2 attenuated CFA-induced mechanical and thermal hyperalgesia and reduced the expression of SDH membrane glutamate receptors (eg mGlu receptor 1, AMPA receptor) and postsynaptic density (eg PSD-95, DLG2). These findings increased the understanding of the role of neurexin-2 in the inflammatory pain, implicating that neurexin-2 acts as a potential regulatory protein of inflammatory pain through affecting synaptic plasticity in the SDH of rats.
Assuntos
Inflamação/complicações , Proteínas do Tecido Nervoso/genética , Dor/etiologia , Dor/metabolismo , Corno Dorsal da Medula Espinal/metabolismo , Animais , Biomarcadores , Modelos Animais de Doenças , Suscetibilidade a Doenças , Adjuvante de Freund/efeitos adversos , Inativação Gênica , Hiperalgesia/diagnóstico , Hiperalgesia/etiologia , Hiperalgesia/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Dor/diagnóstico , Proteoma , Proteômica/métodos , Interferência de RNA , RNA Interferente Pequeno/genética , Ratos , Células Receptoras Sensoriais/metabolismoRESUMO
Descending nociceptive modulation from the supraspinal structures has an important role in cancer-induced bone pain (CIBP). Midbrain ventrolateral periaqueductal gray (vlPAG) is a critical component of descending nociceptive circuits; nevertheless, its precise cellular and molecular mechanisms involved in descending facilitation remain elusive. Our previous study has shown that the activation of p38 MAPK in vlPAG microglia is essential for the neuropathic pain sensitization. However, the existence of potential connection between astrocytes and c-Jun N-terminal kinase (JNK) pathway in CIBP has not yet been elucidated. The following study examines the involvement of astrocyte activation and upregulation of p-JNK in vlPAG, using a CIBP rat model. Briefly, CIBP was mimicked by an intramedullary injection of Walker 256 mammary gland carcinoma cells into the animal tibia. A significant increase in expression levels of astrocytes in the vlPAG of CIBP rats was observed. Furthermore, stereotaxic microinjection of the astrocytic cytotoxin L-α-aminoadipic acid decreased the mechanical allodynia as well as established and reversed the astrocyte activation in CIBP rats. A significant increase in expression levels of p-JNK in astrocytes in vlPAG of CIBP rats was also observed. Moreover, the intrathecal administration of JNK inhibitors SP600125 reduced the expression of glial fibrillary acidic protein, while microinjection of the SP600125 decreased the mechanical allodynia of CIBP rats. These results suggested that CIBP is associated with astrocyte activation in the vlPAG that probably participates in driving descending pain facilitation through the JNK MAPK signaling pathway. To sum up, these findings reveal a novel site of astrocytes modulation of CIBP.
Assuntos
Astrócitos/patologia , Dor do Câncer/patologia , Regulação Neoplásica da Expressão Gênica/fisiologia , MAP Quinase Quinase 4/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Substância Cinzenta Periaquedutal/patologia , Animais , Antracenos/farmacologia , Peso Corporal/efeitos dos fármacos , Neoplasias Ósseas/complicações , Neoplasias Ósseas/patologia , Antígeno CD11b/metabolismo , Dor do Câncer/etiologia , Carcinoma/complicações , Carcinoma/patologia , Linhagem Celular Tumoral , Inibidores Enzimáticos/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Proteína Glial Fibrilar Ácida/metabolismo , Hiperalgesia/etiologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Substância Cinzenta Periaquedutal/metabolismo , Fosfopiruvato Hidratase/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: Despite accumulating evidence on the role of glial cells and their associated chemicals in mechanisms of pain, few studies have addressed the potential role of chemokines in the descending facilitation of chronic pain. We aimed to study the hypothesis that CXCL1/CXCR2 axis in the periaqueductal gray (PAG), a co-restructure of the descending nociceptive system, is involved in descending pain facilitation. METHODS: Intramedullary injection of Walker 256 mammary gland carcinoma cells of adult female Sprague Dawley rats was used to establish a bone cancer pain (BCP) model. RT-PCR, Western blot, and immunohistochemistry were performed to detect pNfkb, Cxcl1, and Cxcr2 and their protein expression in the ventrolateral PAG (vlPAG). Immunohistochemical co-staining with NeuN, GFAP, and CD11 were used to examine the cellular location of pNFκB, CXCL1, and CXCR2. The effects of NFκB and CXCR2 antagonists and CXCL1 neutralizing antibody on pain hypersensitivity were evaluated by behavioral testing. RESULTS: BCP induced cortical bone damage and persistent mechanical allodynia and increased the expression of pNFκB, CXCL1, and CXCR2 in vlPAG. The induced phosphorylation of NFκB was co-localized with GFAP and NeuN, but not with CD11. Micro-injection of BAY11-7082 attenuated BCP and reduced CXCL1 increase in the spinal cord. The expression level of CXCL1 in vlPAG showed co-localization with GFAP, but not with CD11 and NeuN. Micro-administration of CXCL1 neutralizing antibody from 6 to 9 days after inoculation attenuated mechanical allodynia. Furthermore, vlPAG application of CXCL1 elicited pain hypersensitivity in normal rats. Interestingly, CXCR2 was upregulated in vlPAG neurons (not with CD11 and GFAP) after BCP. CXCR2 antagonist SB225002 completely blocked the CXCL1-induced mechanical allodynia and attenuated BCP-induced pain hypersensitivity. CONCLUSION: The NFκB-dependent CXCL1-CXCR2 signaling cascade played a role in glial-neuron interactions and in descending facilitation of BCP.
Assuntos
Astrócitos/metabolismo , Dor do Câncer/patologia , Quimiocina CXCL1/metabolismo , NF-kappa B/metabolismo , Neurônios/metabolismo , Receptores de Interleucina-8B/metabolismo , Analgésicos/uso terapêutico , Animais , Anticorpos/uso terapêutico , Neoplasias Ósseas/complicações , Antígenos CD11/metabolismo , Dor do Câncer/tratamento farmacológico , Dor do Câncer/etiologia , Carcinoma/complicações , Linhagem Celular Tumoral , Quimiocina CXCL1/genética , Quimiocina CXCL1/imunologia , Modelos Animais de Doenças , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Hiperalgesia/etiologia , Hiperalgesia/patologia , NF-kappa B/genética , NF-kappa B/imunologia , Nitrilas/uso terapêutico , Ratos , Ratos Sprague-Dawley , Receptores de Interleucina-8B/genética , Receptores de Interleucina-8B/imunologia , Sulfonas/uso terapêuticoRESUMO
BACKGROUND: Chemokine, monocyte chemoattractant protein-1 (MCP-1), is a potential factor to cause cancer-induced bone pain (CIBP). NF-κB signaling is very important in mediating the expression of chemokines and may have a role in CIBP. However, the mechanism is still unclear. This study investigates the role of NF-κB in CIBP by regulating MCP-1/chemokine CC motif receptor-2 (CCR2) signaling pathway. METHODS: A rat CIBP model was established by injecting Walker-256 cells into the tibia medullary cavity. Nine days later, animals were intrathecally administrated with MCP-1 neutralizing antibody, CCR2 antagonist (RS504393), or NF-кB inhibitor (BAY11-7081). Mechanical paw withdrawal threshold was used to assess pain behavior and sciatic functional index, and radiographic images were adopted to evaluate the damage of nerve and bone. The spinal cords were harvested for Western blot and quantitative reverse transcription polymerase chain reaction. The distribution of MCP-1, CCR2, and NF-кB was detected by double immunofluorescent staining. RESULTS: CIBP caused remarkable bone destruction, injury of sciatic and femoral nerve, and persistent (>15 days) mechanical allodynia in rats. Tumor cell inoculation upregulate MCP-1 and NF-кB in activated neurons as well as CCR2 in neurons and microglia of the spinal cord. MCP-1 antibody, RS504393, and BAY11-7081 partially reversed CIBP-induced mechanical allodynia, and CIBP regulated the expression levels of pro-inflammatory cytokines, tumor necrosis factor-α and interferon-γ, and anti-inflammatory cytokine, interleukin 4, and BAY11-7081 lowered CIBP-induced MCP-1 and CCR2 expressions in a dose-dependent manner. CONCLUSION: In conclusion, NF-кB signaling pathway regulates the expressions of MCP-1/CCR2-induced inflammatory factors in the spinal cord of CIBP rats.
Assuntos
Dor do Câncer/patologia , Quimiocina CCL2/metabolismo , NF-kappa B/metabolismo , Receptores CCR2/metabolismo , Medula Espinal/metabolismo , Animais , Anticorpos/uso terapêutico , Benzoxazinas/uso terapêutico , Neoplasias Ósseas/complicações , Neoplasias Ósseas/diagnóstico por imagem , Dor do Câncer/diagnóstico por imagem , Dor do Câncer/etiologia , Linhagem Celular Tumoral , Quimiocina CCL2/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Nitrilas/uso terapêutico , Limiar da Dor/efeitos dos fármacos , Fosfopiruvato Hidratase/metabolismo , Ratos , Ratos Sprague-Dawley , Compostos de Espiro/uso terapêutico , Sulfonas/uso terapêutico , Fatores de Tempo , Regulação para Cima/efeitos dos fármacosRESUMO
The midbrain ventrolateral periaqueductal gray (VL-PAG) is a key component that mediates pain modulation. Although spinal cord glial cells appear to play an important role in chronic pain development, the precise mechanisms involving descending facilitation pathways from the PAG following nerve injury are poorly understood. This study shows that cellular events that occur during glial activation in the VL-PAG may promote descending facilitation from the PAG during neuropathic pain. Chronic constriction nerve injury (CCI) was induced by ligature construction of the sciatic nerve in male Sprague-Dawley rats. Behavioral responses to noxious mechanical (paw withdrawal threshold; PWT) and thermal (paw withdrawal latency; PWL) stimuli were evaluated. After CCI, immunohistochemical and Western blot analysis of microglia and astrocytes in the VL-PAG showed morphological and quantitative changes indicative of activation in microglia and astrocytes. Intra-VL-PAG injection of microglial or astrocytic inhibitors attenuated PWT and PWL at days 7 and 14, respectively, following CCI. We also evaluated the effects of intra-VL-PAG administration of the phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) inhibitor SB 203580 at day 7 after CCI. This treatment abolished microglial activation and produced a significant time-dependent attenuation of PWT and PWL. Western blot analysis showed localized expression of p-p38 in the VL-PAG after CCI. P-p38 was expressed in labeled microglia of the VL-PAG but was not present in astrocytes and neurons on day 7 after CCI. These results demonstrate that CCI-induced neuropathic pain is associated with glial activation in the VL-PAG, which likely participates in descending pain facilitation through the p38 MAPK signaling pathway.
Assuntos
Neuroglia/patologia , Substância Cinzenta Periaquedutal/patologia , Ciática/patologia , Ciática/fisiopatologia , Transdução de Sinais/fisiologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Anti-Inflamatórios não Esteroides/uso terapêutico , Proteínas de Ligação ao Cálcio/metabolismo , Modelos Animais de Doenças , Proteína Glial Fibrilar Ácida/metabolismo , Hiperalgesia/tratamento farmacológico , Hiperalgesia/fisiopatologia , Imidazóis/uso terapêutico , Masculino , Proteínas dos Microfilamentos/metabolismo , Medição da Dor , Limiar da Dor/efeitos dos fármacos , Fosfopiruvato Hidratase/metabolismo , Piridinas/uso terapêutico , Ratos , Ratos Sprague-Dawley , Ciática/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Fatores de TempoAssuntos
Neoplasias Ósseas/tratamento farmacológico , Neoplasias Ósseas/metabolismo , Dor do Câncer/tratamento farmacológico , Dor do Câncer/metabolismo , Curcumina/análogos & derivados , Curcumina/uso terapêutico , Interleucina-1beta/metabolismo , Janus Quinase 2/metabolismo , Fator de Transcrição STAT3/metabolismo , Animais , Western Blotting , Linhagem Celular Tumoral , Feminino , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosAssuntos
Neoplasias Ósseas/metabolismo , Dor do Câncer/metabolismo , Caspases/metabolismo , Estresse do Retículo Endoplasmático/fisiologia , Corno Dorsal da Medula Espinal/metabolismo , Fator 6 Ativador da Transcrição/genética , Fator 6 Ativador da Transcrição/metabolismo , Animais , Apoptose/genética , Apoptose/fisiologia , Western Blotting , Neoplasias Ósseas/genética , Dor do Câncer/genética , Caspase 3/genética , Caspase 3/metabolismo , Caspases/genética , Estresse do Retículo Endoplasmático/genética , Feminino , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Fosforilação/genética , Fosforilação/fisiologia , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Ratos , Tapsigargina/farmacologiaRESUMO
OBJECTIVE: To explore the roles of P2Y12 receptor (P2Y12R) in bone cancer pain by observing the changes of inflammatory cytokines (IL-1ß, IL-6) after intrathecal injection (i.t.) of P2Y12R antagonist MRS2395. METHODS: Thirty-two female SD rats were randomly divided into 4 groups (n = 8 each): sham group (group S), MRS2395 group (group M), cancer group (group A) and cancer + MRS2395 group (group MA).Groups S and M received an injection of 10 µl Hank's solution into left tibia medullar cavity while groups A and MA had an injection of Walker 256 mammary cancer cells (10 µl, 2×10(7) cells/ml) into the same place. At Day 9-12 post-inoculation, groups S and A received an injection of saline (0.9%, 15 µl, i.t.) while groups M and MA had MRS2395 (400 pmol/µl, 15 µl, i.t.). Intrathecal catheterization between L3 and L4 was performed immediately after inoculating tumor cells by inserting a small tube into vertebral space. Mechanical withdrawal thresholds were measured on left hind paws before and during 10-minute intervals after dosing. Spinal cords (L4-L6 segments) were removed for determining the expressions of IL-1ß and IL-6 by enzyme-linked immunosorbent assay (ELISA) at Day 12 after drug delivery. RESULTS: At 20 min post-injection, mechanical withdrawal thresholds of groups S, M, A and MA were (34.2 ± 5.8), (34.4 ± 5.7), (21.0 ± 2.0) and (25.4 ± 2.3) g respectively at Day 9 post-inoculation (F = 18.679, P < 0.01); mechanical withdrawal thresholds of group A obviously decreased versus groups S and M; mechanical withdrawal thresholds in group MA increased obviously versus group A. The expressions of IL-1ß in groups S, M, A and MA were (74.0 ± 18.6), (98.4 ± 17.3), (253.5 ± 66.4) and (146.3 ± 22.3) pg/ml at Day 12 post-inoculation (F = 18.221, P < 0.01); compared with groups S and M, the expression of IL-1ß in group A showed a significant up-regulation. Likewise, the expressions of IL-6 were (377.4 ± 65.8), (331.6 ± 67.9), (856.1 ± 53.4) and (596.1 ± 34.9) pg/ml (F = 70.880, P < 0.01) respectively in groups S, M, A and MA; compared with groups S and M, the expression of IL-6 increased obviously in group A. There were significant decreases of IL-1ß and IL-6 in group MA versus group A. CONCLUSIONS: An intrathecal injection of MRS2395 may alleviate hyperalgesia and inhibit the up-regulated expression of spinal cord inflammatory cytokines in bone cancer rats. And P2Y12 receptor may be involved in the formation of bone cancer pain through regulating the expressions of IL-1ß and IL-6.
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Neoplasias Ósseas , Dor , Animais , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Hiperalgesia , Injeções Espinhais , Interleucina-1beta , Interleucina-6 , Antagonistas do Receptor Purinérgico P2Y , Ratos , Ratos Sprague-Dawley , Medula Espinal , Regulação para CimaRESUMO
Zoster-associated pain (ZAP) is a painful condition that significantly impacts a patient's quality of life, often leading to postherpetic neuralgia (PHN). Over 30% of patients with herpes probably experience PHN. However, the understanding and treatment of ZAP remain inadequate. Common interventional treatments include radiofrequency therapy, nerve blocks, epidural block, and spinal cord electrical stimulation. Among these, radiofrequency therapy is widely used for pain control in ZAP, but the standard pulsed radiofrequency technique can still be improved. Researchers have explored different radiofrequency parameters, modes, targets, and combined treatments to enhance the therapeutic effect. In this paper, we review the latest research findings and incorporate our own departmental investigations. We conclude that high-voltage, long-duration pulsed radiofrequency and radiofrequency thermocoagulation therapy have shown improved therapeutic outcomes, despite some remaining limitations. Emphasis is placed on safety in intercostal nerve and extracranial nerve radiofrequency treatments. Combination therapy is also safe and effective; however, many studies have a low grade of evidence. Further high-quality research and systematic reviews are needed.
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G protein-coupled receptors (GPCRs), which form the largest family of membrane protein receptors in humans, are highly complex signaling systems with intricate structures and dynamic conformations and locations. Among these receptors, a specific subset is referred to as orphan GPCRs (oGPCRs) and has garnered significant interest in pain research due to their role in both central and peripheral nervous system function. The diversity of GPCR functions is attributed to multiple factors, including allosteric modulators, signaling bias, oligomerization, constitutive signaling, and compartmentalized signaling. This review primarily focuses on the recent advances in oGPCR research on pain mechanisms, discussing the role of specific oGPCRs including GPR34, GPR37, GPR65, GPR83, GPR84, GPR85, GPR132, GPR151, GPR160, GPR171, GPR177, and GPR183. The orphan receptors among these receptors associated with central nervous system diseases are also briefly described. Understanding the functions of these oGPCRs can contribute not only to a deeper understanding of pain mechanisms but also offer a reference for discovering new targets for pain treatment.
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INTRODUCTION: Before tracheal intubation, it is essential to provide sufficient oxygen reserve for emergency patients with full stomachs. Recent studies have demonstrated that high-flow nasal oxygen (HFNO) effectively pre-oxygenates and prolongs apneic oxygenation during tracheal intubation. Despite its effectiveness, the use of HFNO remains controversial due to concerns regarding carbon dioxide clearance. The air leakage and unknown upper airway obstruction during HFNO therapy cause reduced oxygen flow above the vocal cords, possibly weaken the carbon dioxide clearance. METHODS: Patients requiring emergency surgery who had fasted < 8 h and not drunk < 2 h were randomly assigned to the high-flow group, who received 100% oxygen at 30-60 L/min through nasopharyngeal airway (NPA), or the mask group, who received 100% oxygen at 8 L/min. PaO2 and PaCO2 were measured immediately before pre-oxygenation (T0), anesthesia induction (T1), tracheal intubation (T2), and mechanical ventilation (T3). The gastric antrum's cross-sectional area (CSA) was measured using ultrasound technology at T0, T1, and T3. Details of complications, including hypoxemia, reflux, nasopharyngeal bleeding, postoperative pulmonary infection, postoperative nausea and vomiting (PONV), and postoperative nasopharyngeal pain, were recorded. The primary outcomes were PaCO2 measured at T1, T2, and T3. The secondary outcomes included PaO2 at T1, T2, and T3, CSA at T1 and T3, and complications happened during this trial. RESULTS: Pre-oxygenation was administered by high-flow oxygen through NPA (n = 58) or facemask (n = 57) to 115 patients. The mean (SD) PaCO2 was 32.3 (6.7) mmHg in the high-flow group and 34.6 (5.2) mmHg in the mask group (P = 0.045) at T1, 45.0 (5.5) mmHg and 49.4 (4.6) mmHg (P < 0.001) at T2, and 47.9 (5.1) mmHg and 52.9 (4.6) mmHg (P < 0.001) at T3, respectively. The median ([IQR] [range]) PaO2 in the high-flow and mask groups was 404.5 (329.1-458.1 [159.8-552.9]) mmHg and 358.9 (274.0-413.3 [129.0-539.1]) mmHg (P = 0.007) at T1, 343.0 (251.6-428.7 [73.9-522.1]) mmHg and 258.3 (162.5-347.5 [56.0-481.0]) mmHg (P < 0.001) at T2, and 333.5 (229.9-411.4 [60.5-492.4]) mmHg and 149.8 (87.0-246.6 [51.2-447.5]) mmHg (P < 0.001) at T3, respectively. The CSA in the high-flow and mask groups was 371.9 (287.4-557.9 [129.0-991.2]) mm2 and 386.8 (292.0-537.3 [88.3-1651.7]) mm2 at T1 (P = 0.920) and 452.6 (343.7-618.4 [161.6-988.1]) mm2 and 385.6 (306.3-562.0 [105.5-922.9]) mm2 at T3 (P = 0.173), respectively. The number (proportion) of complications in the high-flow and mask groups is shown below: hypoxemia: 1 (1.7%) vs. 9 (15.8%, P = 0.019); reflux: 0 (0%) vs. 0 (0%); nasopharyngeal bleeding: 1 (1.7%) vs. 0 (0%, P = 1.000); pulmonary infection: 4 (6.9%) vs. 3 (5.3%, P = 1.000); PONV: 4 (6.9%) vs. 4 (7.0%, P = 1.000), and nasopharyngeal pain: 0 (0%) vs. 0 (0%). CONCLUSIONS: Compared to facemasks, pre-oxygenation with high-flow oxygen through NPA offers improved carbon dioxide clearance and enhanced oxygenation prior to tracheal intubation in patients undergoing emergency surgery, while the risk of gastric inflation had not been ruled out. TRIAL REGISTRATION: This trial was registered prospectively at the Chinese Clinical Research Registry on 26/4/2022 (Registration number: ChiCTR2200059192).
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Dióxido de Carbono , Intubação Intratraqueal , Máscaras , Oxigenoterapia , Humanos , Masculino , Feminino , Estudos Prospectivos , Pessoa de Meia-Idade , Oxigenoterapia/métodos , Adulto , Oxigênio/metabolismo , Nasofaringe , Respiração Artificial , IdosoRESUMO
The management and therapy of bone cancer pain (BCP) remain formidable clinical challenges. Curcumin and its analogues have been shown to have anti-inflammatory and analgesic properties. In the present study, we investigated the efficacy of curcumin analogue NL04 (NL04) in modulating inflammation in spinal dorsal horn (SDH), thereby exploring its potential to reduce central sensitization of BCP in a rat model. Differing doses of NL04 and curcumin were administered intrathecally either once (on day 12 of BCP) or over seven consecutive days (from day 6-12 of BCP). Results indicated that the ED50 for NL04 and curcumin ameliorating BCP-induced mechanical hyperalgesia is 49.08 µg/kg and 489.6 µg/kg, respectively. The analgesic effects at various doses of NL04 lasted between 4 and 8 h, with sustained administration over a week maintaining pain relief for 1-4 days, while also ameliorating locomotor gait via gait analysis and reducing depressive and anxiety-like behaviors via open-field and light-dark transition tests. The analgesic effects at various doses of curcumin lasted 4 h, with sustained administration over a week maintaining pain relief for 0-2 days. ELISA, Western blotting, qPCR, and immunofluorescence assays substantiated that intrathecal administration of NL04 on days 6-12 of BCP dose-dependently lowered spinal IL-1ß and IL-18 levels and significantly reduced the expression of IKKß genes and proteins, as well as the downstream cleavage of the trans-Golgi network (TGN). Whole-cell patch-clamp results demonstrated that NL04 inhibits potassium ion efflux in rat primary spinal neurons. Thus, NL04 exhibits significant analgesic effects in a BCP rat model by downregulating IKKß expression and inhibiting neuronal potassium ion efflux, which, in turn, suppresses the activation of NLRP3 inflammasomes and reduces IL-1ß production, potentially ameliorating pain management in BCP.
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Neoplasias Ósseas , Dor do Câncer , Curcumina , Ratos , Animais , Dor do Câncer/tratamento farmacológico , Dor do Câncer/metabolismo , Curcumina/farmacologia , Curcumina/uso terapêutico , Curcumina/metabolismo , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Sensibilização do Sistema Nervoso Central , Quinase I-kappa B/metabolismo , Dor/tratamento farmacológico , Neoplasias Ósseas/complicações , Neoplasias Ósseas/tratamento farmacológico , Neoplasias Ósseas/metabolismo , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Analgésicos/metabolismo , Hiperalgesia/tratamento farmacológico , Hiperalgesia/metabolismo , Medula Espinal , Potássio/metabolismoRESUMO
Background: Bone cancer pain (BCP) represents one of the most challenging comorbidities associated with cancer metastasis. Long non-coding RNAs (lncRNAs) have garnered attention as potential therapeutic agents in managing neuropathic pain. However, their role in the regulation of nociceptive information processing remains poorly understood. In this study, we observed a significant down-regulation of the spinal lncRNA ENSRNOG00000051325 (lncRNA51325) in a rat model of bone cancer pain. Our study sought to elucidate the potential involvement of lncRNA51325 in the development of BCP by modulating the expression of molecules associated with pain modulation. Methods: We established the BCP model by injecting Walker 256 cells into the tibial plateau of rats. We conducted tests on the pain behaviors and anxiety-like responses of rats through von-Frey test, Gait analysis, and Open Field Test. Spinal lumbar expansion was harvested for molecular biology experiments to explore the relationship between lncRNA51325 and Pumilio RNA binding family member 2 (Pum2). Results: Notably, the overexpression of lncRNA51325 effectively attenuated mechanical allodynia in rats afflicted with BCP, whereas the knockdown of lncRNA51325 induced pain behaviors and anxiety-like responses in naïve rats. Additionally, we observed a time-dependent increase in the expression of Pum2 in BCP-afflicted rats, and intrathecal injection of Pum2-siRNA alleviated hyperalgesia. Furthermore, our investigations revealed that lncRNA51325 exerts a negative modulatory effect on Pum2 expression. The overexpression of lncRNA51325 significantly suppressed Pum2 expression in BCP rats, while the knockdown of lncRNA51325 led to elevated Pum2 protein levels in the spinal cord of naïve rats. Subsequent treatment with Pum2-siRNA mitigated the downregulation of lncRNA51325-induced mechanical allodynia in naïve rats. Conclusion: Our findings indicate that lncRNA51325 plays a role in regulating bone cancer pain by inhibiting Pum2 expression, offering a promising avenue for novel treatments targeting nociceptive hypersensitivity induced by bone metastatic cancer.