ABSTRACT
Oxycodone has been used clinically for over 90 years. While it is known that it exhibits low affinity for the multiple opioid receptors, whether its pharmacological activities are due to oxycodone activation of the opioid receptor type or due to its active metabolite (oxymorphone) that exhibits high affinity for the mu-opioid receptors remains unresolved. Ross and Smith (1997) reported the antinociceptive effects of oxycodone (171nmol, i.c.v.) are induced by putative kappa-opioid receptors in SD rat while others have reported oxycodone activities are due to activation of mu- and/or delta-opioid receptors. In this study, using male mu-opioid receptor knock-out (MOR-KO) mice, we examined whether delta-opioid receptor was involved in oxycodone antinociception. Systemic subcutaneous (s.c.) administration of oxycodone (above 40mg/kg) could induce a small but significant antinociceptive effect in MOR-KO mice by the tail flick test. Delta-opioid receptor antagonist (naltrindole, 10mg/kg or 20mg/kg, i.p.) could block this effect. When oxycodone was injected directly into the brain of MOR-KO mice by intracerebroventricular (i.c.v.) route, oxycodone at doses of 50nmol or higher could induce similar level of antinociceptive responses to those observed in wild type mice at the same doses by i.c.v. Delta-opioid receptor antagonists (naltrindole at 10nmol or ICI 154,129 at 20µg) completely blocked the supraspinal antinociceptive effect of oxycodone in MOR-KO mice. Such oxycodone antinociceptive responses were probably not due to its active metabolites oxymorphone because (a) the relative low level of oxymorphone was found in the brain after systemically or centrally oxycodone injection using LC/MS/MS analysis; (b) oxymorphone at a dose that mimics the level detected in the mice brain did not show any significant antinocieption effect; (c) oxycodone exhibits equal potency as oxymorphone albeit being a partial agonist in regulating [Ca(2+)]I transients in a clonal cell line expressing high level of mu-opioid receptor. These data suggest that oxycodone by itself can activate both the mu- and delta-opioid receptors and that delta-opioid receptors may contribute to the central antinociceptive effect of oxycodone in mice.
Subject(s)
Analgesics, Opioid/pharmacology , Brain/drug effects , Nociception/drug effects , Nociceptive Pain/prevention & control , Oxycodone/pharmacology , Pain Threshold/drug effects , Receptors, Opioid, delta/agonists , Analgesics, Opioid/administration & dosage , Animals , Brain/metabolism , Brain/physiopathology , Disease Models, Animal , Dose-Response Relationship, Drug , Genotype , Injections, Intraventricular , Injections, Subcutaneous , Male , Mice, Inbred C57BL , Mice, Knockout , Narcotic Antagonists/pharmacology , Nociceptive Pain/genetics , Nociceptive Pain/metabolism , Nociceptive Pain/physiopathology , Oxycodone/administration & dosage , Phenotype , Receptors, Opioid, delta/metabolism , Receptors, Opioid, mu/genetics , Receptors, Opioid, mu/metabolism , Signal Transduction/drug effects , Time FactorsABSTRACT
Denervation-mediated skeletal muscle atrophy results from the loss of electric stimulation and leads to protein degradation, which is critically regulated by the well-confirmed transcriptional co-activator peroxisome proliferator co-activator 1 alpha (PGC-1α). No adequate treatments of muscle wasting are available. Pyrroloquinoline quinone (PQQ), a naturally occurring antioxidant component with multiple functions including mitochondrial modulation, demonstrates the ability to protect against muscle dysfunction. However, it remains unclear whether PQQ enhances PGC-1α activation and resists skeletal muscle atrophy in mice subjected to a denervation operation. This work investigates the expression of PGC-1α and mitochondrial function in the skeletal muscle of denervated mice administered PQQ. The C57BL6/J mouse was subjected to a hindlimb sciatic axotomy. A PQQ-containing ALZET® osmotic pump (equivalent to 4.5 mg/day/kg b.w.) was implanted subcutaneously into the right lower abdomen of the mouse. In the time course study, the mouse was sacrificed and the gastrocnemius muscle was prepared for further myopathological staining, energy metabolism analysis, western blotting, and real-time quantitative PCR studies. We observed that PQQ administration abolished the denervation-induced decrease in muscle mass and reduced mitochondrial activities, as evidenced by the reduced fiber size and the decreased expression of cytochrome c oxidase and NADH-tetrazolium reductase. Bioenergetic analysis demonstrated that PQQ reprogrammed the denervation-induced increase in the mitochondrial oxygen consumption rate (OCR) and led to an increase in the extracellular acidification rate (ECAR), a measurement of the glycolytic metabolism. The protein levels of PGC-1α and the electron transport chain (ETC) complexes were also increased by treatment with PQQ. Furthermore, PQQ administration highly enhanced the expression of oxidative fibers and maintained the type II glycolytic fibers. This pre-clinical in vivo study suggests that PQQ may provide a potent therapeutic benefit for the treatment of denervation-induced atrophy by activating PGC-1α and maintaining the mitochondrial ETC complex in skeletal muscles.
Subject(s)
Mitochondria/metabolism , Muscle, Skeletal/drug effects , PQQ Cofactor/pharmacology , Transcription Factors/metabolism , Animals , Denervation , Electron Transport , Mice , Mice, Inbred C57BL , Muscle, Skeletal/innervation , Muscle, Skeletal/metabolism , Muscle, Skeletal/pathology , Oxidative Phosphorylation , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alphaABSTRACT
BACKGROUND: Neuropathic pain is a very troublesome and difficult pain to treat. Although opioids are the best analgesics for cancer and surgical pain in clinic, only oxycodone among opioids shows better efficacy to alleviate neuropathic pain. However, many side effects associated with the use of oxycodone render the continued use of it in neuropathic pain treatment undesirable. Hence, we explored whether dextromethorphan (DM, a known N-methyl-D-aspartate receptor antagonist with neuroprotective properties) could potentiate the anti-allodynic effect of oxycodone and underlying mechanisms regarding to glial cells (astrocytes and microglia) activation and proinflammatory cytokines release in a spinal nerve injury (SNL) mice model. RESULTS: Oxycodone produced a dose-dependent anti-allodynic effect. Co-administration of DM at a dose of 10 mg/kg (i.p.) (DM10) which had no anti-allodynic effect by itself enhanced the acute oxycodone (1 mg/kg, s.c.) effect. When the chronic anti-allodynic effects were examined, co-administration of DM10 also significantly enhanced the oxycodone effect at 3 mg/kg. Furthermore, oxycodone decreased SNL-induced activation of glial cells (astrocytes and microglia) and plasma levels of proinflammatory cytokines (IL-6, IL-1ß and TNF-α). Co-administration of DM10 potentiated these effects of oxycodone. CONCLUSION: The combined use of DM with oxycodone may have therapeutic potential for decreasing the effective dose of oxycodone on the treatment of neuropathic pain. Attenuation of the glial activation and proinflammatory cytokines in the spinal cord may be important mechanisms for these effects of DM.
Subject(s)
Dextromethorphan/pharmacology , Neuralgia/drug therapy , Oxycodone/pharmacology , Animals , Astrocytes/metabolism , Astrocytes/pathology , Cytokines/metabolism , Dextromethorphan/agonists , Disease Models, Animal , Drug Synergism , Male , Mice , Microglia/metabolism , Microglia/pathology , Neuralgia/metabolism , Neuralgia/pathology , Oxycodone/agonistsABSTRACT
OBJECTIVE: This study was designed to demonstrate that alendronate (ALN)/vitamin D3 combination tablets (ALN/D5600) are bioequivalent to corresponding doses of ALN and vitamin D3 as individual tablets in healthy Taiwanese volunteers. METHODS: In this open-label, randomized, 2-period, crossover study, 68 volunteers were randomized to a single ALN/D5600 combination tablet or corresponding doses of 70 mg ALN + 5600 IU vitamin D3 (2 × 2800 IU), followed by a 12-day washout period and administration of the alternate formulation. Plasma ALN levels were measured using a newly developed assay. Geometric mean ratios of ALN AUC0-last, AUC0-∞, and Cmax, and unadjusted vitamin D3 AUC0-80h and Cmax were compared and considered bioequivalent if the 90% CI was within 0.8 to 1.25. RESULTS: The geometric mean ratios were: AUC0-last, 1.084 (90% CI, 0.937-1.253); AUC0-∞, 1.081 (90% CI, 0.935-1.249); and Cmax, 1.112 (90% CI, 0.959-1.289) for ALN, and AUC0-80h 0.953 (90% CI, 0.827-1.098) and Cmax, 0.982 (90% CI, 0.854-1.130) for vitamin D3 unadjusted for endogenous levels. CONCLUSIONS: The combination tablet was considered bioequivalent to coadministration based on ALN AUC0-∞ and unadjusted vitamin D3 parameters. Slight differences for ALN AUC0-last and Cmax (upper 90% CIs outside the bounds) were not considered clinically significant. The combination tablet was well tolerated. No serious adverse experiences were reported. © 2015. The Authors. Published by Elsevier Inc. All rights reserved.
ABSTRACT
BACKGROUND: Glycogen-synthase kinase 3 (GSK3) is involved in many signaling pathways and is associated with a host of high-profile pathophysiological states. However, its role in morphine tolerance, especially naloxone-precipitated withdrawal syndrome, has not been well investigated. The present study was undertaken to study the role of GSK3 in chronic morphine exposure. METHODS: Adult male Sprague-Dawley rats were subjected to intraperitoneal (i.p.) injections of morphine (10 mg/kg) twice daily for 6 consecutive days, and tail-flick tests were conducted to evaluate changes of morphine-induced antinociception. GSK3 inhibitor, SB216763 or SB415286, was i.p. injected prior to morphine to investigate the influences on morphine tolerance. There were four groups receiving morphine plus vehicle (2% dimethyl sulfoxide), morphine plus SB216763 (0.6 mg/kg) or SB415286 (1.0 mg/kg), GSK3 inhibitor alone, or dimethyl sulfoxide: as the control group. On Day 7, naloxone (i.p., 1 mg/kg) was administered and naloxone-precipitated withdrawal behaviors were individually compared between groups. RESULTS: Repeated morphine exposure in this study led to progressive shortening of tail-flick latencies and produced six of nine observed naloxone-precipitated withdrawal behaviors. Coadministration with SB216763 or SB415286 significantly prevented antinociceptive tolerance and alleviated parts of withdrawal syndrome. Both inhibitors could similarly reverse withdrawal behaviors including grooming, chewing, and ptosis, but did not affect withdrawal behaviors of penis licking and defecation. CONCLUSION: The results demonstrate the importance of GSK3 in reducing chronic morphine-induced tolerance and withdrawal syndrome. Although GSK3 is involved in diverse physiological functions, aiming at GSK3-related pathway could still be a potential tool to improve therapeutic quality in clinical morphine treatment.
Subject(s)
Drug Tolerance/physiology , Glycogen Synthase Kinase 3/antagonists & inhibitors , Morphine/administration & dosage , Protein Kinase Inhibitors/administration & dosage , Aminophenols/administration & dosage , Animals , Drug Combinations , Glycogen Synthase Kinase 3/physiology , Indoles/administration & dosage , Male , Maleimides/administration & dosage , Rats , Rats, Sprague-Dawley , Substance Withdrawal Syndrome/enzymologyABSTRACT
BACKGROUND: Cognitive impairment has been documented in adult diabetes but is unclear in pediatric diabetes. No study had been conducted to explore the relationship between attention-deficit/hyperactivity disorder (ADHD) and diabetes. Using a population-based data set, we aimed to examine the association between ADHD and a prior diagnosis of diabetes mellitus (DM) in Taiwan. METHODS: A total of 4,302 patients with ADHD were selected as cases and 21,510 randomly selected subjects as controls. We used conditional logistic regression to calculate the odds ratio (OR) for having previously received a diagnosis of DM between subjects with and without ADHD. RESULTS: In this study, 116 of the 25,812 sampled subjects (0.5%) had received a diagnosis of DM prior to their index date. Subjects with ADHD had a higher proportion of prior DM diagnoses than controls (0.9% vs. 0.4%, P < 0.001). After adjusting for age, sex, index year, geographic location, and obesity, ADHD was significantly associated with a prior diagnosis of type 2 DM (OR = 2.75, 95% confidence interval (CI) = 1.82-4.16). However, no significant association was observed between ADHD and type 1 DM. CONCLUSION: The findings suggest that ADHD was associated with a previous diagnosis of type 2 DM.
Subject(s)
Attention Deficit Disorder with Hyperactivity/epidemiology , Diabetes Mellitus, Type 2/epidemiology , Adolescent , Attention Deficit Disorder with Hyperactivity/diagnosis , Case-Control Studies , Child , Child, Preschool , Diabetes Mellitus, Type 1/epidemiology , Diabetes Mellitus, Type 2/diagnosis , Female , Humans , Logistic Models , Male , Odds Ratio , Prevalence , Risk Factors , Taiwan/epidemiologyABSTRACT
Kleine-Levin syndrome (KLS) is a rare disorder characterized by recurrent episodes of hypersomnia, cognitive or behavior disturbances, compulsive eating behavior, and hypersexuality. The etiology of KLS remains unknown even though its clinical symptoms suggest an underlying autoimmune process. In this study, we analyzed the human leukocyte antigen (HLA) typing alleles in Taiwanese patients with KLS using the polymerase chain reaction sequence-specific priming technique. We report that an immunoresponsive HLA-DQB1, DQB1∗0602, was detected in significant quantities in patients with KLS (three of 12, p=0.046) and could elevate the risk of KLS (odds ratio, 1.143; 95% confidence interval, 0.0982-1.329). In conclusion, an identification of genomic susceptibility to KLS will be helpful in determining the immunospecific targeted therapies for patients with KLS.
Subject(s)
Genetic Predisposition to Disease/genetics , HLA-DQ beta-Chains/genetics , Kleine-Levin Syndrome/genetics , Adolescent , Alleles , Child , Female , Genotype , Humans , Male , Polymerase Chain Reaction , Taiwan , Young AdultABSTRACT
The study was to examine the effects of Sanguis draconis ethanol extract (SDEE) on streptozotocin (STZ)- and cytokine-induced ß-cell damage. In vitro, SDEE did not cause cytotoxicity below 200 µg/ml, and can prevent STZ (5mM)-induced cell death and apoptosis below 100 µg/ml on RIN-m5F cells. SDEE inhibits IL-1ß/IFN-γ-stimulated NO, TNF-α release, and iNOS expression. Furthermore, SDEE suppressed the IL-1ß/IFN-γ- or STZ-induced p65 expression of NF-κB, which is associated with inhibition of IκB-α degradation. In vivo, treatment of ICR mice with STZ (100 mg/kg, i.p. single injection) resulted in hyperglycemia and hypoinsulinemia, which was further evidenced by blood glucose and plasma insulin. The diabetogenic effects of STZ were completely prevented when mice were orally administered with SDEE for 3 weeks, however, the blood glucose and plasma insulin showed no significant change after SDEE administration alone. In addition, SDEE also can inhibit STZ-induced iNOS protein expression, pancreatic injury and lipid peroxidation. In conclusions, the molecular mechanism by which SDEE inhibits iNOS gene expression appears to involve the inhibition of NF-κB activation. These results suggest the possible therapeutic value of S. draconis and could be potentially developed into a novel drug for preventing the progression of diabetes mellitus.
Subject(s)
Insulin-Secreting Cells/drug effects , Insulin-Secreting Cells/pathology , Insulinoma/drug therapy , Interferon-gamma/toxicity , Interleukin-1beta/toxicity , Plant Extracts/pharmacology , Streptozocin/toxicity , Animals , Antibiotics, Antineoplastic/toxicity , Antiviral Agents/toxicity , Blood Glucose , Blotting, Western , Cell Survival/drug effects , Cytosol/metabolism , Hyperglycemia/chemically induced , Hyperglycemia/drug therapy , In Vitro Techniques , Insulin-Secreting Cells/metabolism , Islets of Langerhans/drug effects , Lipid Peroxidation/drug effects , Male , Mice , Mice, Inbred ICR , Nitric Oxide/metabolism , Rats , Tumor Necrosis Factor-alpha/metabolismABSTRACT
The aim of this study was to evaluate the effect of curcumin on morphine tolerance and the corresponding cytokine/chemokine changes. Male ICR mice were made tolerant to morphine by daily subcutaneous injection for 7 days. Intraperitoneal injections of vehicle, low-dose or high-dose curcumin were administered 15 min after morphine injection, either acutely or chronically for 7 days to test the effect of curcumin on morphine-induced antinociception and development of morphine tolerance. On day 8, cumulative dose-response curves were generated and the 50% of maximal analgesic dose values were calculated and compared among groups. Corresponding set of mice were used for analyzing the cytokine responses by antibody-based cytokine protein array. Acute, high-dose curcumin enhanced morphine-induced antinociception. While morphine tolerance was attenuated by administration of low-dose curcumin following morphine injections for 7 days, it was aggravated by chronic high-dose curcumin following morphine injection, suggesting a biphasic effect of curcumin on morphine-induced tolerance. Of the 96 cytokine/chemokines analyzed by mouse cytokine protein array, 14 cytokines exhibited significant changes after the different 7-day treatments. Mechanisms for the modulatory effects of low-dose and high-dose curcumin on morphine tolerance were discussed. Even though curcumin itself is a neuroprotectant and low doses of the compound serve to attenuate morphine tolerance, high-doses of curcumin might cause neurotoxicity and aggravate morphine tolerance by inhibiting the expression of antiapoptotic cytokines and neuroprotective factors. Our results indicate that the effect of curcumin on morphine tolerance may be biphasic, and therefore curcumin should be used cautiously.
ABSTRACT
OBJECTIVES: To elucidate a novel anti-inflammatory mechanism of myrrh against lipopolysaccharide (LPS)-induced inflammation. METHODS: RAW264.7 macrophages were cultured in DMEM and then cells were treated with LPS or LPS plus a myrrh methanol extract (MME) for 24h. The culture medium was collected for determination of nitric oxide (NO), prostaglandin (PG)E(2) , interleukin (IL)-1ß, and tumour necrosis factor (TNF)-α, and cells were harvested by lysis buffer for Western blot analysis. KEY FINDINGS: Our data showed that treatment with the MME (1â¼100µg/ml) did not cause cytotoxicity or activate haem oxygenase-1 (HO-1) protein synthesis in RAW264.7 macrophages. Furthermore, the MME inhibited LPS-stimulated NO, PGE(2) , IL-1ß and TNF-α release and inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 protein expression. Zn(II) protoporphyrin IX, a specific inhibitor of HO-1, blocked the inhibition of iNOS and COX-2 expression by the MME. CONCLUSIONS: These results suggest that among mechanisms of the anti-inflammatory response, the MME inhibited the production of NO, PGE(2) , IL-1ß and TNF-α by downregulating iNOS and COX-2 gene expression in macrophages and worked through the action of HO-1.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Commiphora , Heme Oxygenase-1/biosynthesis , Inflammation Mediators/metabolism , Inflammation/metabolism , Phytotherapy , Plant Extracts/pharmacology , Animals , Anti-Inflammatory Agents/therapeutic use , Inflammation/drug therapy , Lipopolysaccharides , Macrophages/drug effects , Mice , Plant Extracts/therapeutic useABSTRACT
Maternal methamphetamine (MA) abuse during pregnancy has been proved to induce various impacts on the development of infant and child. In this study, we examined whether prenatal exposure to MA would affect the development of nociceptive system by measuring the responses to noxious stimulation in the developing rat. Adult female Sprague-Dawley rats received bi-daily subcutaneous injection of methamphetamine (5mg/kg) or isovolumetric normal saline since the day of mating till the day of delivery. Birth profiles of the offspring including birth length, weight, and body temperature were recorded during the first postnatal month. Mechanical withdrawal thresholds were measured by von Frey filaments on postnatal day (PND) 30 and 60, and hyperalgesic behaviors following plantar formalin injection (2%, 50 microl) were evaluated on PND 60. The birth body weight and length of rats born to MA-injected dam rats (MA group) were significantly lower than those of the control rats during the first postnatal month; however, their body temperature was significantly higher than those of the control rats during the first 3 days after birth. The MA group rats had significantly lower tactile withdrawal values in von Frey test and higher pain scores in the late phase of pain in the formalin test than those of the control rats. There is a gender difference of nociceptive hypersensitivity manifested as that the female MA group rats had significantly lower withdrawal thresholds and higher pain scores in response to formalin injection than the male MA group rats. These results suggest that prenatal MA exposure could predispose an alteration in the development of nociceptive neuronal network, which leads to a long-lasting status of hypersensitivity to pain stimulations in the offspring.
Subject(s)
Hyperalgesia/physiopathology , Methamphetamine , Pain Threshold/drug effects , Prenatal Exposure Delayed Effects/chemically induced , Prenatal Exposure Delayed Effects/physiopathology , Age Factors , Analysis of Variance , Animals , Animals, Newborn , Birth Weight , Body Temperature/drug effects , Body Weight/drug effects , Disease Models, Animal , Female , Formaldehyde/adverse effects , Hyperalgesia/chemically induced , Male , Methamphetamine/pharmacology , Pain Measurement/methods , Pregnancy , Rats , Rats, Sprague-Dawley , Reaction Time/drug effects , Sex CharacteristicsABSTRACT
BACKGROUND: Diffuse noxious inhibitory controls (DNIC) can be produced by different types of conditioning stimuli, but the analgesic properties and underlying mechanisms remain unclear. The aim of this study was to differentiate the induction of DNIC analgesia between noxious electrical and inflammatory conditioning stimuli. METHODS: First, rats subjected to either a supramaximal electrical stimulation or an injection of high-dose formalin in the hind limb were identified to have pain responses with behavioral evidence and spinal Fos-immunoreactive profiles. Second, suppression of tail-flick latencies by the two noxious stimuli was assessed to confirm the presence of DNIC. Third, an opioid receptor antagonist (naloxone) and an alpha2-adrenoreceptor antagonist (yohimbine) were injected, intraperitoneally and intrathecally respectively, before conditioning noxious stimuli to test the involvement of descending inhibitory pathways in DNIC-mediated analgesia. RESULTS: An intramuscular injection of 100 microl of 5% formalin produced noxious behaviors with cumulative pain scores similar to those of 50 microl of 2% formalin in the paw. Both electrical and chemical stimulation significantly increased Fos expression in the superficial dorsal horns, but possessed characteristic distribution patterns individually. Both conditioning stimuli prolonged the tail-flick latencies indicating a DNIC response. However, the electrical stimulation-induced DNIC was reversed by yohimbine, but not by naloxone; whereas noxious formalin-induced analgesia was both naloxone- and yohimbine-reversible. CONCLUSIONS: It is demonstrated that DNIC produced by different types of conditioning stimuli can be mediated by different descending inhibitory controls, indicating the organization within the central nervous circuit is complex and possibly exhibits particular clinical manifestations.
Subject(s)
Afferent Pathways/physiopathology , Analgesia/methods , Neural Inhibition/physiology , Pain/physiopathology , Adrenergic alpha-Antagonists/pharmacology , Afferent Pathways/drug effects , Analysis of Variance , Animals , Area Under Curve , Conditioning, Psychological/physiology , Electric Stimulation , Formaldehyde/administration & dosage , Immunohistochemistry , Male , Naloxone/pharmacology , Narcotic Antagonists , Neural Inhibition/drug effects , Rats , Rats, Sprague-Dawley , Yohimbine/pharmacologyABSTRACT
The aging population is a global phenomenon. The skyrocketing costs of healthcare and the shortage of healthcare providers will soon become a crucial issue all over the world. Taiwan's government executed the Taiwan's Telehealth Pilot Project (TTPP) from July 1, 2008 to December 31, 2008, using healthcare information technology to tackle these problems. The system has three different models, the home-care, the community-care, and the residential-care model to assist the elderly in the pursuit of better healthcare and improved quality of life. The results revealed both the home-care and community-care models facilitated timely medical responses if the enrolled patients had emergent conditions. In the home-care model, the hospital readmission rate was reduced from 8.19% to 3.17%, and the hospital visit rate was decreased from 2.95% to 2.90%. In community-care model, the medication nonadherence rate was reduced from 38.20% to 9.20%. In the residential-care model, reduced rates of readmission to the hospital, nosocomial infection and the adverse drug event were found. Telehealth enabled the aged with chronic illnesses to live independently and helped the institutionalized elderly get acute care more efficiently without increased manpower of healthcare organization.
Subject(s)
Long-Term Care , Telemedicine , Pilot Projects , TaiwanABSTRACT
BACKGROUND: Recent studies have implicated the activation of stress-activated mitogen-activated protein kinase (MAPK) p38 in spinal microglial cells for development of neuropathic and inflammatory pain. The aim of the present study was to investigate whether phosphorylation of p38 (p-p38) also mediates mechanical allodynia and thermal hyperalgesia induced by plantar incision. METHODS: After rats received a plantar incision surgery, mechanical allodynia and thermal hyperalgesia were determined by von Frey filaments and radiant heat, respectively, and the number of p-p38 immunoreactive cells in the dorsal horn was quantified to determine p38 activation at different time points after incision. The p38 inhibitor FR167653 was administered intrathecally 30 min before hind paw plantar incision to determine the role of p38 in postoperative pain. RESULTS: A significant increase in number of p-p38 immunoreactive cells was observed in the ipsilateral L4-5 spinal dorsal horn from 1 h to 3 days after the incision. p-p38 was found predominantly in microglia. However, microglial activation (assessed by OX-42 upregulation) was not evident until 3 days after plantar incision. Intrathecal pretreatment of FR167653 attenuated incision-induced mechanical allodynia from 1 h to day 2 and significantly reduced activation of p38 in the dorsal horn 1 day after plantar incision. However, FR167653 only inhibited heat hyperalgesia at an early time point. CONCLUSIONS: Plantar incision-induced mechanical allodynia can be prevented by the p38 inhibitor. Our results suggest that p38 activation in spinal microglia play a role in incision-induced mechanical allodynia in rats. Therefore, p38 inhibition may be useful in treating postsurgical pain.
Subject(s)
Microglia/enzymology , Pain Measurement/methods , Pain/enzymology , Spinal Cord/enzymology , p38 Mitogen-Activated Protein Kinases/metabolism , Animals , Enzyme Activation/drug effects , Enzyme Activation/physiology , Male , Microglia/drug effects , Pain/prevention & control , Pain Measurement/drug effects , Pain Measurement/instrumentation , Physical Stimulation/methods , Pyrazoles/pharmacology , Pyrazoles/therapeutic use , Pyridines/pharmacology , Pyridines/therapeutic use , Rats , Rats, Sprague-Dawley , Spinal Cord/drug effects , p38 Mitogen-Activated Protein Kinases/antagonists & inhibitorsABSTRACT
The use of anesthetics in acupuncture analgesia is controversial. We evaluate a steady-state light anesthesia model to test whether minimal stress manipulation and reliable measurement of analgesia could be simultaneously achieved during electroacupuncture (EA) in animals. A series of experiments were performed. Firstly, EA compliance and tail-flick latencies (TFL) were compared in rats under 0.1%, 0.3%, 0.5%, 0.7%, or 1.1% halothane for 120min. Under 0.5% halothane, TFL were then measured in groups receiving EA at intensity of 3, 10 or 20 volt (V), 1 or 2mg/kg morphine, 20V EA plus naloxone, or control. Subsequently, the effect of EA on formalin-induced hyperalgesia was tested and c-fos expression in the spinal dorsal horn was analyzed. Rats exhibited profound irritable behaviors and highly variable TFL under 0.1% or 0.3% halothane, as well as a time-dependent increase of TFL under 0.7% or 1.1% halothane. TFL remained constant at 0.5% halothane, and needle insertion and electrical stimulation were well tolerated. Under 0.5% halothane, EA increased TFL and suppressed formalin-induced hyperalgesia in an intensity-dependent and naloxone-reversible manner. EA of 20V prolonged TFL by 74%, suppressed formalin-induced hyperalgesia by 32.6% and decreased c-fos expression by 29.7% at the superficial and deep dorsal horn with statistically significant difference. In conclusion, 0.5% halothane provides a steady-state anesthetic level which enables the humane application of EA stimulus with the least interference on analgesic assessment. This condition serves as a minimal stress EA model in animals devoid of stress-induced analgesia while maintaining physiological and biochemical response in the experiment.
Subject(s)
Acupuncture Analgesia/methods , Disease Models, Animal , Electroacupuncture , Pain Threshold , Rats, Sprague-Dawley , Stress, Physiological/prevention & control , Anesthetics, Inhalation/pharmacology , Animals , Halothane/pharmacology , Hyperalgesia/therapy , Male , Neurons/metabolism , Pain Measurement , Proto-Oncogene Proteins c-fos/metabolism , Rats , Reaction Time/physiology , TailABSTRACT
Methamphetamine (MA) is well known as a potent CNS stimulant, which produces strong rewarding and behavioral sensitization after repeated administration. In the present study, we investigated whether co-administration of dextromethorphan (DM) with MA could suppress these effects induced by acute and chronic MA treatment. The conditioned place preference (CPP) test was used to examine the rewarding/drug seeking effects and locomotor and stereotypic activities were measured to investigate behavioral sensitization induced by chronic MA. Our results revealed that co-administration of DM (20 mg/kg, ip) with MA (2 mg/kg, ip) almost completely abolished the MA-induced CPP and behavioral sensitization. Furthermore, both of the acute and chronic MA could result in an increase of dopamine (DA) turnover rate in the NAc and mPFC. The acute effects of MA on DA turnover rate could be attenuated by the co-administration of DM in both regions. The chronic effect of MA on DA turnover rate in the mPFC was also attenuated by the co-administration of DM. These results suggest that the effect of DM on blocking MA-induced rewarding and behavioral sensitization may be related to its effect on inhibiting the activity of DA neurons projected to mPFC and/or NAc.
Subject(s)
Behavior, Animal/drug effects , Central Nervous System Stimulants/antagonists & inhibitors , Dextromethorphan/pharmacology , Excitatory Amino Acid Antagonists/pharmacology , Methamphetamine/antagonists & inhibitors , Amphetamine-Related Disorders/metabolism , Animals , Central Nervous System Stimulants/pharmacology , Dopamine/metabolism , Male , Methamphetamine/pharmacology , Nucleus Accumbens/metabolism , Prefrontal Cortex/metabolism , Rats , Rats, Sprague-Dawley , RewardABSTRACT
In this study, we first demonstrated that loratadine (LOR), a promising world widely used oral anti-histamine, effectively inhibits growth of tumors derived from human colon cancer cells (COLO 205) in an in vivo setting. In vitro study demonstrated that the anti-tumor effects of LOR in COLO 205 cells were mediated by causing G(2)/M phase cell growth cycle arrest and caspase 9-mediated apoptosis. Cell-cycle arrest induced by LOR (75 microM, 24 h) was associated with a significant decrease in protein levels of cyclin B1, cell division cycle (Cdc) 25B, and Cdc25C, leading to accumulation of Tyr-15-phosphorylated Cdc2 (inactive form). Interestingly, LOR (75 microM, for 4 h) treatment also resulted in a rapid and sustained phosphorylation of Cdc25C at Ser-216, leading to its translocation from the nucleus to the cytoplasm because of increased binding with 14-3-3. We further demonstrated that the LOR-induced Cdc25C (Ser-216) phosphorylation was blocked in the presence of checkpoint kinase 1 (Chk1) specific inhibitor (SB-218078). The cells treated with LOR in the presence of Chk1 specific inhibitor (SB-218078) were then released from G(2)/M arrest into apoptosis. These results implied that Chk1-mediated phosphorylation of Cdc25C plays a major role in response to LOR-mediated G(2)/M arrest. Although the Chk1-mediated cell growth arrest in response to DNA damage is well documented, our results presented in this study was the first report to describe the Chk1-mediated G(2)/M cell-cycle arrest by the histamine H1 antagonist, LOR.
Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cell Cycle Proteins/metabolism , Cell Division/drug effects , G2 Phase/drug effects , Loratadine/pharmacology , Protein Kinases/metabolism , cdc25 Phosphatases/metabolism , Adenocarcinoma , Cell Cycle/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Checkpoint Kinase 1 , Colonic Neoplasms , Cytosol/drug effects , Cytosol/physiology , Humans , Mitochondria/drug effects , Mitochondria/physiology , Phosphorylation , Protease Inhibitors/pharmacologyABSTRACT
Infants passively exposed to morphine or heroin through their addicted mothers usually develop characteristic withdrawal syndrome of morphine after birth. In such early life, the central nervous system exhibits significant plasticity and can be altered by various prenatal influences, including prenatal morphine exposure. Here we studied the effects of prenatal morphine exposure on postsynaptic density protein 95 (PSD-95), an important cytoskeletal specialization involved in the anchoring of the NMDAR and neuronal nitric oxide synthase (nNOS), of the hippocampal CA1 subregion from young offspring at postnatal day 14 (P14). We also evaluated the therapeutic efficacy of dextromethorphan, a widely used antitussive drug with noncompetitive antagonistic effects on NMDARs, for such offspring. The results revealed that prenatal morphine exposure caused a maximal decrease in PSD-95 expression at P14 followed by an age-dependent improvement. In addition, prenatal morphine exposure reduced not only the expression of nNOS and the phosphorylation of cAMP responsive element-binding protein at serine 133 (CREB(Serine-133)), but also the magnitude of long-term depression (LTD) at P14. Subsequently, the morphine-treated offspring exhibited impaired performance in long-term learning and memory at later ages (P28-29). Prenatal coadministration of dextromethorphan with morphine during pregnancy and throughout lactation could significantly attenuate the adverse effects as described above. Collectively, the study demonstrates that maternal exposure to morphine decreases the magnitude of PSD-95, nNOS, the phosphorylation of CREB(Serine-133), and LTD expression in hippocampal CA1 subregion of young offspring (e.g., P14). Such alterations within the developing brain may play a role for subsequent neurological impairments (e.g., impaired performance of long-term learning and memory). The results raise a possibility that postsynaptic density proteins could serve an important role, at least in part, for the neurobiological pathogenesis in offspring from the morphine-addicted mother and provide tentative therapeutic strategy.
Subject(s)
Dextromethorphan/pharmacology , Hippocampus/drug effects , Intracellular Signaling Peptides and Proteins/drug effects , Membrane Proteins/drug effects , Morphine Dependence/complications , Morphine/adverse effects , Prenatal Exposure Delayed Effects/metabolism , Animals , Animals, Newborn , Cyclic AMP Response Element-Binding Protein/drug effects , Cyclic AMP Response Element-Binding Protein/metabolism , Dextromethorphan/therapeutic use , Disease Models, Animal , Disks Large Homolog 4 Protein , Excitatory Amino Acid Antagonists/pharmacology , Excitatory Amino Acid Antagonists/therapeutic use , Female , Hippocampus/metabolism , Hippocampus/physiopathology , Intracellular Signaling Peptides and Proteins/metabolism , Long-Term Synaptic Depression/drug effects , Long-Term Synaptic Depression/physiology , Membrane Proteins/metabolism , Narcotics/adverse effects , Neuronal Plasticity/drug effects , Neuronal Plasticity/physiology , Nitric Oxide Synthase Type I/drug effects , Nitric Oxide Synthase Type I/metabolism , Organ Culture Techniques , Phosphorylation/drug effects , Pregnancy , Prenatal Exposure Delayed Effects/drug therapy , Prenatal Exposure Delayed Effects/physiopathology , Rats , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/drug effects , Receptors, N-Methyl-D-Aspartate/metabolism , Synaptic Membranes/drug effects , Synaptic Membranes/metabolismABSTRACT
UNLABELLED: Pediatric populations are at risk for medication errors that may be associated with mortality and disability. The purpose of this study was to describe the clinical manifestations of seven newborns following an event of accidental intramuscular injection of atracurium and to assess the impact on neurodevelopmental outcome. This study enrolled seven newborns who were accidentally administered 10 mg of atracurium, equivalent to 2.6-3.3 mg/kg, in a local perinatal clinic. Accident reports and hospital records were reviewed to obtain the history and medical data for the event. The survivors were prospectively examined for their growth, health and neurodevelopment until 24 months of age. All newborns showed immediate apnea and cyanosis requiring resuscitation after atracurium injection and presented with respiratory failure and flaccid paralysis on arrival for emergency medical services. One newborn was asystolic despite resuscitation and died of multiple organ failure. Of the five survivors available for follow-up, all achieved favorable growth outcomes. However, four showed mild to significant delay in development; and two manifested mild hypomyelination of cerebral white matter on the brain magnetic resonance imaging. CONCLUSION: Newborns accidentally injected with high doses of atracurium are at risk of death and neurodevelopmental delay. The serious clinical manifestations, developmental delay and cerebral hypomyelination were most likely due to insufficient immediate respiratory assistance following atracurium injection.
Subject(s)
Atracurium/poisoning , Brain/drug effects , Developmental Disabilities/chemically induced , Medical Errors , Neuromuscular Nondepolarizing Agents/poisoning , Atracurium/administration & dosage , Bradycardia/chemically induced , Brain/pathology , Cyanosis/chemically induced , Drug Overdose , Female , Follow-Up Studies , Humans , Hypoxia/chemically induced , Infant, Newborn , Injections, Intramuscular , Magnetic Resonance Imaging , Male , Motor Skills/drug effects , Neuromuscular Nondepolarizing Agents/administration & dosage , Nurseries, HospitalABSTRACT
Previous investigations had shown that inhibitor of serotonin reuptake transporter (SERT) could attenuate morphine withdrawal syndrome in adult animals. In the present study, we determined whether postnatal injection of serotonin reuptake inhibitors, fluoxetine, clomipramine, or citalopram, is able to attenuate the expression of the naloxone-precipitated morphine withdrawal syndrome in 5-day-old neonatal Sprauge-Dawley rats born to dams rat that received morphine injection since a week before mating till 5 days after delivery. Withdrawal syndrome of morphine, manifested as frequent abdominal stretching and yawning, was generated by injection of naloxone on postnatal day 5. Pre-injection with fluoxetine, clomipramine, or citalopram, significantly attenuated the naloxone-precipitated syndrome in a dose-dependent manner without apparent side effect. The rank order of inhibitory potency is citalopram=clomipramine>fluoxetine. This result suggests that inhibitor of SERT may be of potential in treating neonatal morphine withdrawal syndrome.