Cytokines in the central nervous system: targets for therapeutic intervention

Accumulating evidence implicates inflammatory processes in the development of a number of neurodegenerative diseases and demonstrates that neurons and microglia can be a source for various cytokines, which are believed to be involved in neuropathology, and therefore can serve as targets for therapeutic treatment. Moreover, it is now established that many of these pro-inflammatory molecules, commonly associated with the peripheral immune system, are also produced within the central nervous system (CNS). The term 'cytokine network' has been widely used to describe cytokine biology in the brain. However, the function of this network has not been well-characterised. It is believed that understanding the function of this network might have important clinical applications. This article reviews recent and current developments in cytokine research that pertain to the development of new strategies targeting cytokines in the brain, thus opening up new avenues for novel therapeutic approaches for the treatment of various pathological conditions and diseases of the CNS.     

Inhibition of glutathione-related enzymes augments LPS-mediated cytokine biosynthesis: involvement of an IkappaB/NF-kappaB-sensitive pathway in the alveolar epithelium

The regulation of lipopolysaccharide (LPS)-mediated pro-inflammatory cytokine biosynthesis by reduction-oxidation (redox)-sensitive enzymes involved in maintaining intracellular glutathione homeostasis was investigated in fetal alveolar type II epithelial cells (fATII). Inhibition of glutathione-oxidized disulfide reductase, which recycles GSSG --> 2GSH, by the action of 1,3-bis-(2-chloroethyl)-1-nitrosourea (BCNU) augmented LPS-dependent secretion of interleukin (IL)-1beta, IL-6 and tumor necrosis factor (TNF)-alpha. BCNU increased [GSSG] concentration at the expense of [GSH], thereby favoring oxidation equilibrium. Inhibition of gamma-glutamylcysteine synthetase, the rate-limiting enzyme in the biosynthesis of GSH, by the action of L-buthionine-(S,R)-sulfoximine (BSO), potentiated LPS-induced IL-1beta, IL-6 and TNF-alpha production. Similar to BCNU, BSO depleted [GSH] and induced the accumulation of [GSSG]. BCNU and BSO reduced LPS-mediated phosphorylation of inhibitory-kappaB (IkappaB-alpha), allowing its cytosolic accumulation. This effect was associated with the inhibition of the nuclear translocation of selective nuclear factor (NF)-kappaB subunits: NF-kappaB1 (p50), RelA (p65), RelB (p68) and c-Rel (p75), but not NF-kappaB2 (p52). BCNU and BSO reduced LPS-induced NF-kappaB activation as determined by the electrophoretic mobility shift DNA-binding assay. Analytical analysis of the effect of modulating the dynamic redox ratio ([GSH]+[GSSG])/[GSSG] revealed a novel role for GSSG as a disulfhydryl compound which mediates an inhibitory effect on NF-kappaB activation. It is concluded that selective modulation of redox-sensitive enzymes has an immunopharmacological potential in regulating pro-inflammatory cytokines and that the TkappaB-alpha/NF-kappaB pathway is redox-sensitive and differentially involved in mediating redox-dependent regulation of cytokine signaling.     

The role of cytokines and prostaglandin-E(2) in thymulin induced hyperalgesia

We have recently reported that intraperitoneal (i.p.) injection of thymulin at low doses (50 ng) resulted in thermal and mechanical hyperalgesia and upregulation of the level of interleukin-1beta in the liver. In this study, we demonstrate that such injections of thymulin result in a significant elevation in the levels of TNF-alpha (P<0.01), NGF (P<0.01) and PGE(2) (P<0.01) in the liver of the treated rats, in addition to the increase in the levels of IL-1beta. Pretreatment with specific antagonists to each of these factors (polyclonal anti-TNF-alpha, anti-NGF antiserum and IL-1 receptor antagonist) did not result in the abolition of the hyperalgesia as assessed by the paw pressure, hot plate, paw immersion and tail flick tests. However, pretreatment with a combination of the above antagonist and antisera almost completely prevented thymulin-induced hyperalgesia. The cyclooxygenase inhibitor, meloxicam, reversed in a dose dependent manner (0.2, 0.4 and 2 mg/kg) thymulin effects as assessed by the different pain tests. It also abolished the thymulin-induced increase in the level of cytokines and NGF in the liver. Our results indicate that PGE(2) could be the key mediator of the hyperalgesic action of thymulin and the observed upregulation of proinflammatory cytokines and NGF.     

A single administration of the hallucinogen, 4‐acetoxy‐dimethyltryptamine,prevents the shift to a drug‐dependent state and the expression of withdrawal aversions in rodents

Despite several studies suggesting the therapeutic use of 5‐hydroxytryptamine receptors type 2A (5‐HT_2A) agonists in the treatment of substance use disorders, the neurobiological basis accounting for such effects are still unknown. It has been observed that chronic exposure to drugs of abuse produces molecular and cellular adaptations in ventral tegmental area (VTA) neurons, mediated by brain‐derived neurotrophic factor (BDNF). These BDNF‐induced adaptations in the VTA are associated with the establishment of aversive withdrawal motivation that leads to a drug‐dependent state. Growing evidence suggests that 5‐HT_2A receptor signaling can regulate the expression of BDNF in the brain. In this study, we observed that a single systemic or intra‐VTA administration of a 5‐HT_2A agonist in rats and mice blocks both the aversive conditioned response to drug withdrawal and the mechanism responsible for switching from a drug‐naive to a drug‐dependent motivational system. Our results suggest that 5‐HT_2A agonists could be used as therapeutic agents to reverse a drug dependent state, as well as inhibiting the aversive effects produced by drug withdrawal.     

The role of the sympathetic efferents in endotoxin-induced localized inflammatory hyperalgesia and cytokine upregulation

The sympathetic system (SNS) is considered to be a major component of the neurogenic contribution to inflammation and hyperalgesia. We have investigated the role of the SNS in the local inflammatory pain induced by intraplantar (i.pl) injections of bacterial endotoxin (ET). Treatment of rats with an alpha-adrenoceptor antagonist (phentolamine, 0.25-1 mg/kg, i.p.), a beta-adrenoceptor antagonist (propranolol, 1-10 mg/kg, p.o.) or a sympathetic neuron-blocking agent (guanethedine, 30 mg/kg, s.c.) resulted in a dose-dependent reduction of the thermal hyperalgesia induced by ET. Mechanical hyperalgesia, however, was less sensitive to inhibition by propranolol and guanethedine but significantly inhibited by phentolamine. ET injection produced significant upregulation of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), IL-6, and nerve growth factor (NGF). Treatment with any one of the three sympatholytics abolished the upregulation of NGF and IL-6, while phentolamine and guanethedine also reversed the upregulation of TNF-alpha. IL-1 beta was resistant to all of the sympatholytic treatments. We conclude that the SNS can contribute to the local inflammation and hyperalgesia following injection of ET. The resistance to sympatholytics shown by IL-1 beta, known to play a key role in the inflammatory cascade, suggests that ET can initiate inflammation and hyperalgesia independently of peripheral and central sympathetic mechanisms.     

Macrophage recruitment and activation: a model for comparing resistance to Salmonella enteritidis in different broiler breeds

A model for comparing resistance to Salmonella Enteritidis was evaluated in different broiler breeds. The recruitment and phagocytic activity of peritoneal macrophages were assessed in three different broiler breeds (A, B and C) which are farmed world-wide. Assessment was performed after three days of intraperitoneal (i.p.) administration of 3% Sephadex G-200 (10 ml), initiated at twenty-one days of age, followed by contact with i.p. live S. Enteritidis (10 ml, 1.2 x 10(8) colony forming units/ml) for 45 min. Assessment included determination of the number of i.p. macrophages recruited, the number of i.p. phagocytized S. Enteritidis cells per macrophage, the levels of degranulated i.p. beta-glucuronidase and beta-galactosidase, and the count of surviving S. Enteritidis cells. Confirmation of the significance of the model was obtained by comparing resistance to field infection by S. Enteritidis in the three broiler breeds. The recruitment of i.p. macrophages in response to challenge with Sephadex and S. Enteritidis was significantly higher (P < 0.05) in birds of breed A (mean cumulative i.p. macrophage count, in 10 fields of microscopic slide smear magnified at x1,000, was equal to 81.7), compared to recruitment in birds of breed B (33.3) or breed C (41.2). The mean number of phagocytized S. Enteritidis cells per i.p. macrophage in birds of breed A (2.68) was significantly higher (P < 0.05) than in breed B (0.83) and insignificantly higher (P > 0.05) than in breed C (2.35). In addition, the highest level of recruitment and phagocytic activity of macrophages, in birds of breed A, was associated with a higher significant mean i.p. beta-glucuronidase activity (10,425.5 units/ml) than in breed B (3,438.2 units/ml) or breed C (3,356.94 units/ml) (P < 0.05). Moreover, birds of breed A demonstrated a higher mean i.p. beta-galactosidase activity (2.225 units/ml) than birds of breed B (0.852 units/ml) or breed C (1.852 units/ml) (P > 0.05). The higher level of recruitment and activity of i.p. macrophages and the higher rate of degranulation of i.p. enzymes in breed A were associated with a greater number of surviving i.p. S. Enteritidis cells. In response to outbreaks of S. Enteritidis in the field, the average mortality was significantly higher in flocks of breed A (3.2%) than in flocks of breed B (1.2%) or breed C (0.96%) (P < 0.05). These data provide an indication of the significance of the model in reflecting the differences in resistance of S. Enteritidis of broiler breeds reared in a farm environment.     

Fos-like immunoreactivity induced by intraplantar injection of endotoxin and its reduction by morphine

C-Fos-like immunoreactivity (FLI) in the central nervous system, has been associated with the processing of nociceptive information in acute and chronic pain animal models. The aim of this study was to investigate whether intraplantar (i.pl.) injections of endotoxin (ET, 1.25 μg) can induce FLI in the lumbar spinal cord of rats and to assess the effects of morphine injection on c-fos expression. FLI was studied in various groups of rats at 2, 3, 4, 6, 9 and 24 h following ET injections. Labeled neurons were mainly detected in the lumbar segments ipsilateral to the ET-injected leg, with a major peak (71.01±4.79 positive neurons) at 4 h and a second peak (29.87±5.97 positive neurons) at 9 h followed by a recovery to the baseline at 24 h after ET injections. Within the laminae, the majority of positive neurons was observed at 2–3 h in laminae I and II and in deep laminae (V and VI mainly) starting at 4 h after ET injections. Rostrocaudally, labeled neurons were observed initially in L₄–L₅ segments (2–3 h post-ET) after which they extended to L₂–L₆ segments at 4 h after ET. Morphine injections either i.p. (1 or 2 mg/kg) or i.pl. (50 μg) significantly reduced ET-induced hyperalgesia and simultaneously the FLI. The maximum effect was observed on labeled neurons in the deep laminae (V and VI mainly). We conclude that local injections of ET can induce FLI in the lumbar spinal cord with a temporal and spatial patterns comparable to the described hyperalgesia, and that both FLI and hyperalgesia are reduced by morphine in a dose-dependent manner with a maximal effect shown by the local i.pl. morphine injections.