Research reportSpinal orexin-1 receptors mediate anti-hyperalgesic effects of intrathecally-administered orexins in diabetic neuropathic pain model rats
Introduction
Diabetic neuropathy is one of the three most common complications of diabetes and occurs predominantly in the distal extremities as pain, which can occur either spontaneously, as a result of exposure to only mildly painful stimuli (i.e., hyperalgesia) or to stimuli not normally perceived as painful (i.e., allodynia) [3], [9]. The condition remains difficult to treat, because it is relatively resistant to standard analgesics such as opioids [2], [9].
Orexin-A and orexin-B, also known as hypocretin-1 and hypocretin-2, are composed of 33 and 28 amino acids, respectively. They are novel hypothalamic neuropeptides derived from the same 130-amino acid precursor peptide, prepro-orexin [25]. Orexins act on their targets via 2 types of G-protein-coupled receptors; orexin-1 receptors, which have an approximately 10-fold greater affinity for orexin-A than orexin-B, and orexin-2 receptors, which have a similar affinity for both [14], [25]. Their link with nociception has been extended, as studies have demonstrated that activation of the orexinergic system can produce an antinociceptive reaction in experimental animals. For example, in a rat inflammatory pain model caused by formalin injection, orexin-1 receptors were shown to mediate the analgesic action produced by intrathecal orexin-A [31]. Furthermore, our previous study [27] and one by Yamamoto et al. [33] demonstrated that intrathecal injection of orexin-A produced profound analgesic and antiallodynic effects, respectively, in the sciatic nerve of mono-neuropathic pain model rats. In addition, in a rat model of postoperative pain, both orexin-A and orexin-B were shown to induce mild but distinct antiallodynic effects when given intrathecally [8]. However, the effects of orexins on diabetic neuropathic pain have not been investigated.
Several lines of evidence suggest that the orexin-expressing neurons themselves are regulated by energy balance, while orexin expression has been shown to be stimulated by hypoglycemia in fasted rats [25]. Hypothalamic prepro-orexin mRNA levels have also been shown to be increased under both prolonged fasting [4], [25] and insulin-induced hypoglycemic [16] conditions and reduced under hyperglycemic conditions in genetically obese mice [30]. Furthermore, in studies that used fos-like immunoreactivity as a marker of neuronal activation, it has been reported that hypothalamic orexin-containing neurons are activated by insulin-induced hypoglycemia [5], [23]. Thus, orexin-expressing neurons may be a part of a group of glucose-sensitive neurons. In addition, immunohistochemical staining of orexin-containing neurons in a recent experiment demonstrated that orexins may be directly secreted into the cerebrospinal fluid [7]. Therefore, it is conceivable that orexin levels in the cerebrospinal fluid may be reduced in diabetic rats because of hyperglycemia.
In the central nervous system, orexins are localized in the narrow regions within the lateral hypothalamus and then projected to many regions involved with the spinal cord [29], while recent mapping of orexin fibers in rat brain and spinal cord tissues has revealed a differential distribution of the 2 peptides [12]. In the spinal cord dorsal horn, orexin-containing fibers are predominantly present in the superficial laminae associated with nociceptive processing [13], [17], [29]. Furthermore, detailed mapping of orexin receptor mRNA distribution by in situ hybridization has shown that the 2 orexin receptors are distributed throughout the rat brain, with different expression patterns [19], [21], [28]. In addition, both orexin-A and orexin-1 receptors have been found in dorsal root ganglion cells of the spinal cord [1]. These results strongly suggest that the orexinergic system is important for the regulation of different physiological functions, including nociception, and may have a potential role in the modulation of nociceptive transmission, especially in pain pathways.
In the present study, we investigated the effects of intrathecal orexins on both mechanical and thermal hyperalgesia induced by diabetes. Orexin levels in cerebrospinal fluid samples might be more indicative of orexin neurotransmission activity than orexin levels in brain homogenate samples, since cerebrospinal fluid orexin-A levels remain very stable and the measurement method is reliable [15]. We measured orexin levels in rat cerebrospinal fluid samples using a radioimmunoassay method to test our hypothesis that cerebrospinal fluid orexin levels are reduced in diabetic rats because of hyperglycemia. The functional roles of spinal orexin-1 receptors on the anti-hyperalgesic effects of orexins were also examined following intrathecal pretreatment with SB-334867, a selective orexin-1 receptor antagonist, to investigate the underlying mechanisms.
Section snippets
Materials and methods
The experimental protocols used were approved by our institutional animal care committee and were consistent with the guidelines of the ethical committee of the International Association for the Study of Pain [34]. Four hundred six adult male Sprague–Dawley rats, initially weighing 220–250 g, were used. All animals were individually housed at a constant room temperature of 23 ± 2 °C under a 12-h light–dark cycle (lights on at 8:00 am) throughout the course of the study.
Results
Nearly all (97%) of the rats developed hyperglycemia within 1 week after streptozotocin treatment. The plasma glucose concentrations for the diabetic and age-matched normal rats were 388 ± 11 and 99 ± 7 mg/dl (P < 0.05), respectively. Paw-withdrawal threshold before streptozotocin treatment was 58.8 ± 2.6 g in all rats, while the mechanical threshold was decreased significantly (21.2 ± 2.3 g; P < 0.05) at 3 weeks after streptozotocin treatment in the diabetic rats. The sustained
Discussion
In the present study, we demonstrated that intrathecal administration of orexin-A at 1–30 μg produced profound anti-mechanical and thermal hyperalgesic effects, without detectable side effects, in a dose-dependent manner in diabetic neuropathic pain model rats. In addition, intrathecally administered orexin-B at a low dose of 1–3 μg produced no significant effect on the levels of diabetes-induced mechanical and thermal hyperalgesia, in contrast to a high dose between 10 and 30 μg, following
Acknowledgment
We gratefully appreciate the generous gift of SB-334867 from GlaxoSmithKline (Harlow, Essex. UK).
References (34)
- et al.
Orexin-A, an hypothalamic peptide with analgesic properties
Pain
(2001) - et al.
Diabetic neuropathy
Med. Clin. North Am.
(1998) - et al.
Orexin A-like immunoreactivity in the rat brain
Neurosci. Lett.
(1999) - et al.
Streptozocin-induced diabetic rats: behavioural evidence for a model of chronic pain
Pain
(1993) - et al.
Study of the sensitivity of the diabetes-induced pain model in rats to a rage of analgesics
Pain
(1994) - et al.
Differential distribution of orexin-A and orexin-B immunoreactivity in the rat brain and spinal cord
Peptides
(1999) - et al.
Distribution of orexin-A and orexin-B (hypocretins) in the rat spinal cord
Neurosci. Lett.
(2000) - et al.
Insulin-induced hypoglycemia increases preprohypocretin (orexin) mRNA in the rat lateral hypothalamic area
Neurosci. Lett.
(1999) - et al.
Gene expression and protein distribution of the orexin-1 receptor in the rat brain and spinal cord
Neuroscience
(2001) - et al.
Streptozotocin-induced diabetes selectively alters the potency of analgesia produced by mu-opioid agonists, but not by delta- and kappa-opioid agonists
Brain Res.
(1992)
Differential distribution and regulation of OX1 and OX2 orexin/hypocretin receptor messenger RNA in the brain upon fasting
Horm. Behav.
A pharmacologic analysis of mechanical hyperalgesia in streptozotocin/diabetic rats
Pain
Neurons containing orexin in the lateral hypothalamic area of the adult rat brain are activated by insulin-induced acute hypoglycemia
Neurosci. Lett.
1,3-biarylureas as selective non-peptide antagonists of the orexin-1 receptor
Bioorg. Med. Chem. Lett.
Orexin and orexin receptors: a family of hypothalamic neuropeptides and G protein-coupled receptors that regulate feeding behavior
Cell
Distribution of orexin receptor mRNA in the rat brain
FEBS Lett.
Down regulation of the prepro-orexin gene expression in genetically obese mice
Brain Res.
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2020, NeuropeptidesCitation Excerpt :Specifically, the intrathecal administration of orexins reduces neuropathic hyperalgesia in rats with streptozotocin-induced diabetes, whereas it does not influence pain sensitivity in normal rats (Kajiyama et al., 2005; Niknia et al., 2019). The antinociceptive action of orexins in diabetic rats could be abolished by pretreatment with SB-334867 – a selective OX1R antagonist (Kajiyama et al., 2005). Two studies point to the role of OXA in the mouse model of stress-induced analgesia.
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2018, NeuropeptidesCitation Excerpt :Carbachol (250 nM/0.5 μl, n = 7, and 500 nM/0.5 μl; n = 6) or saline (n = 6) was microinjected into the LH, and effect of this stimulation on nociceptive behaviors in both early and late phases was compared to those in vehicle group which received intra-LH administration of saline (0.5 μl). In next step, to examine the effect of intrathecal administration of OX1R antagonist on the LH-induced antinociception, experimental groups were intrathecally microinjected with SB-334867 (3 μM; n = 5, 10 μM; n = 6, 30 μM; n = 5 and 100 μM; n = 5) (Kajiyama et al., 2005), or DMSO (n = 6) 5 min prior to carbachol microinjection (250 nM/0.5 μl) into the LH. DMSO-control group received intrathecally DMSO 12% (10 μl) as drug vehicle before intra-LH administration of carbachol (250 nM/0.5 μl).