Glucocorticoid Receptor Contributes to Electroacupuncture-Induced Analgesia by Inhibiting Nav1.7 Expression in Rats With Inflammatory Pain Induced by Complete Freund's Adjuvant

BackgroundWhile electroacupuncture (EA) has been used traditionally for the treatment of chronic pain, its analgesic mechanisms have not been fully clarified. We observed in an earlier study that EA could reverse inflammatory pain and suppress high Nav1.7 expression. However, the molecular mechanism underlying Nav1.7 expression regulation is unclear. In this study, we studied the relationship between the glucocorticoid receptor (GR) and Nav1.7 and the role of these molecules in EA analgesia.Materials and MethodsIn this study, we established an inflammatory pain model by intraplantar injection of complete Freund's adjuvant (CFA) in rats. EA stimulation was applied to the ipsilateral “Huantiao” (GB30) and “Zusanli” (ST36) acupoints in the rat model. Western blotting, real-time polymerase chain reaction, immunostaining, intrathecal injection, and chromatin immunoprecipitation (ChIP) assay were performed to determine whether the sodium channel protein Nav1.7 plays a role in CFA-induced pain and whether GR regulates Nav1.7 expression during analgesia following EA stimulation.ResultsEA application significantly decreased the paw withdrawal threshold thresholds and thermal paw withdrawal latency and suppressed GR and Nav1.7 expression in the dorsal root ganglion. Moreover, treatment with a GR sense oligonucleotide (OND) markedly reversed these alterations. In contrast, treatment with a GR antisense OND along with EA application exerted a better analgesic effect, which was accompanied by the suppression of Nav1.7 and GR protein expression. The ChIP assay showed that the binding activity of GR to the Nav1.7 promoter was enhanced in CFA injected rats and suppressed in EA-treated rats.ConclusionsThe present study demonstrated that EA exerted anti-hyperalgesic effects by inhibiting GR expression, which led to Nav1.7 expression modulation in the rat model of CFA-induced inflammatory pain.     

Levodopa analgesia in experimental neuropathic pain

Levodopa has been shown to produce analgesia in various clinical and experimental settings, but its use for chronic pain treatment has not been established. We have undertaken a study of the antiallodynic actions of levodopa in a rat model of painful mononeuropathy. When administered systemically, levodopa produced a decrease in tactile and cold allodynia lasting at least 3 h. Direct intrathecal (i.t.) levodopa injection at lumbar levels produced a similar, though shorter, antiallodynic effect. This effect was blocked by the D2-type receptor antagonist sulpiride, which supports the involvement of the spinal dopaminergic system in the analgesic action of levodopa on neuropathic pain. These results provide experimental support on the antiallodynic effect of levodopa in neuropathic pain and suggest that at least part of the analgesic action takes place in the spinal cord and involves dopaminergic D2-type receptors.     

Failure of Placebo Analgesia Model in Rats with Inflammatory Pain

With the shifting role of placebos,there is a need to develop animal models of placebo analgesia and elucidate the mechanisms underlying the effect.In the present study,male Sprague-Dawley rats with chronic inflammatory pain caused by complete Freund’s adjuvant(CFA)underwent a series of conditioning procedures,in which morphine was associated with different cues,but they failed to induce placebo analgesia.Then,conditioning with the conditioned place preference apparatus successfully induced analgesic expectancy and placebo analgesia in naive rats but only induced analgesic expectancy and no analgesic effect in CFA rats.Subsequently,we found enhanced c-fos expression in the nucleus accumbens and reduced expression in the anterior cingulate cortex in naive rats while c-fos expression in the anterior cingulate cortex in CFA rats was not altered.In summary,the behavioral conditioning model demonstrated the difficulty of establishing a placebo analgesia model in rats with a pathological condition.     

Extracellular signal-regulated kinase,substance P and neurokinin-1 are involved in the analgesic mechanism of herb-partitioned moxibustion

Herb-partitioned moxibustion can effectively mitigate visceral pain, a major symptom in inflammatory bowel disease, but the analgesic mechanism is still unclear. Moreover, extracellular signal-regulated kinase, substance P, and neurokinin-1 are involved in formation of central hyperalgesia. Thus, we postulated that the analgesic effect of herb-partitioned moxibustion may be associated with these factors. Accordingly, in this study, we established an inflammatory bowel disease visceral pain model in rat by enema with a mixed solution of 5%trinitrobenzenesulfonic acid and 50% ethanol. BilateralTianshu (ST25) andQihai (CV6) points were selected for herb-partitioned moxi-bustion. Our results showed that herb-partitioned moxibustion improved visceral pain and down-regulated extracellular signal-regulated kinase, substance P, and neurokinin-1 protein and mRNA expression in dorsal root ganglia. These results indicate that down-regulation of extracellular signal-regulated kinase, substance P, and neurokinin-1 protein and mRNA may be a central mechanism for the analgesic effect of herb-partitioned moxibustion.     

Baclofen as an analgesic in chronic peripheral nerve disease

Baclofen has shown analgesic properties in a number of animal studies but has failed as a conventional analgesic in the human postoperative dental pain model. In order to test baclofen's analgesic properties in more chronic pain conditions, we selected postherpetic neuralgia and diabetic neuropathy pain as possible trial diseases for baclofen analgesia. 15 patients with postherpetic neuralgia and 10 with diabetic neuropathy pain were treated with baclofen. In the spinal postherpetic neuralgia group and diabetic neuropathy group, there was little evidence of analgesic effect. 6 of 7 patients with facial postherpetic neuralgia had a good response to baclofen during the 3-week trial. Baclofen does not appear to be a conventional analgesic.     

The Analgesic Effects of Intrathecally Pumped Saline and Artificial Cerebrospinal Fluid in a Rat Model of Neuropathic Pain.

Objective. This experiment was performed to test the hypothesis that intrathecally pumped saline, but not artificial cerebrospinal fluid (CSF), would be analgesic in a rat model of neuropathic pain. Materials and Methods. Surgery for the chronic constriction injury (CCI) model of neuropathic pain and intrathecal catheter placement were performed on the rats, baseline pain testing and pump implantation were performed 7 days later, and pain tests were repeated on days 1, 4, 7, and 14 after pump implantation. Results. Intrathecally pumped saline and artificial CSF were analgesic for cold allodynia (p < 0.05), and intrathecally pumped saline but not CSF for heat nociception in the affected paw (p < 0.005) compared to rats with unattached subcutaneous pumps. No analgesia was observed on tests of spontaneous pain or pressure hyperalgesia (p > 0.1). Conclusions. Intrathecally pumped saline and artificial CSF have analgesic effects on some neuropathic and normal, nociceptive pain signs in CCI rats.     

Activation of central mu-opioid receptors is involved in clonidine analgesia in rats

The analgesic effect of clonidine in spontaneously hypertensive rats (SHR) and in normotensive Sprague-Dawley (SD) rats was assessed by using the formalin pain test. The analgesic response of SD rats to low doses (15-60 micrograms/kg i.p.) but not to a high dose (150 micrograms/kg i.p.) of clonidine was inhibited by naloxone, 2 mg/kg i.p., and similar interaction was noted in SHR. In both rat strains, the analgesic response to low i.p. doses of clonidine was also inhibited by injection of 5 micrograms of naloxone or 7 micrograms of beta-funaltrexamine, a mu-receptor antagonist, into the lateral cerebral ventricle. I.c.v. injection of 5 micrograms of ICI 174864, a delta-receptor antagonist, potentiated or did not influence clonidine analgesia in SD rats and SHR, respectively. It is concluded that the analgesic response to clonidine involves activation of central mu-opioid receptors in both SHR and SD rats, possibly by an endogenous opioid released by clonidine.     

Effect of intrathecal nocistatin on nociceptin/orphanin FQ analgesia in chronic constriction injury rat

Nocistatin and nociceptin/orphanin FQ (N/OFQ) are two neuropeptides derived from the same precursor protein, prepronociceptin (ppOFQ), and exhibit different effects on spinal neurotransmission. Nocistatin does not bind to nociceptin/orphanin FQ peptide receptor (NOP), but intrathecal (i.t.) nocistatin has been reported to block the analgesic effect of i.t. N/OFQ. In this study, we investigated the effect of i.t. nocistatin on N/OFQ analgesia to radiant thermal stimuli in chronic constriction injury (CCI) rat. Firstly, to investigate the analgesic effect of N/OFQ, different doses of N/OFQ (3, 10, 30 microg) were intrathecally injected and foot withdrawal latency (FWL) to radiant heat was recorded. It is observed that 3 microg N/OFQ had no effect on FWL, 10 and 30 microg N/OFQ significantly increased FWL of CCI rat. Then, 10 microg N/OFQ, 10 microg nocistatin and a drug cocktail including 10 microg N/OFQ and 10 microg nocistatin were intrathecally injected. The results showed that FWL significantly decreased after using N/OFQ and nocistatin compared with using only N/OFQ, and 10 microg nocistatin had no effect on FWL versus control, suggesting that this dose of nocistatin per se had no effect on the pain threshold of CCI rat, but could block the analgesic effect of N/OFQ. These results indicated that i.t. N/OFQ dose-relatedly depressed thermal hyperalgesia produced by CCI and nocistatin could block N/OFQ analgesia at spinal level in CCI rat.     

Reversible tetracaine block of rat periaqueductal gray (PAG) decreases baseline tail-flick latency and prevents analgesic effects of met-enkephalin injections in nucleus paragigantocellularis (PGC)

One μg of tetracaine in the rat periaqueductal gray (PAG) produced adecline in baseline tail-flick latencies (hyperalgesia) from about 5 to 3.5 s over the course of 9 min, after which the latencies increased to about 4.5 s. One μg of Met-enkephalin in PGC caused an expected increase in latencies (analgesia) from about 4.25 to 6.2 s in 9 min, with recovery to 4.7 s after 15 min post-injection. Giving the preceding 2 nanoinjectionssimultaneously led to an essentially total block of the PGC analgesia. A control injection in PAG simultaneous with a Met-enkephalin injection in PGC didnot block the latter's analgesic effect. Single control (artificial cerebrospinal fluid) injections in PAG or PGC were without effect. The hyperalgesic effect of PAG tetracain supports the involvement of PAG in normally occuring, tonic descending pain inhibition. The block of PGC met-enkephalin analgesia by distant injection of tetracaine in PAG supports the necessity of PAG integrity for PGC analgesic function.     

Implication of delta opioid receptor subtype 2 but not delta opioid receptor subtype 1 in the development of morphine analgesic tolerance in a rat model of chronic inflammatory pain

Opioids are well known for their robust analgesic effects. Chronic activation of mu opioid receptors (MOPs) is, however, accompanied by various unwanted effects such as analgesic tolerance. Among other mechanisms, interactions between MOPs and delta opioid receptors (DOPs) are thought to play an important role in morphine‐induced behavioral adaptations. Interestingly, certain conditions such as inflammation enhance the function of the DOP through a MOP‐dependent mechanism. Here, we investigated the role of DOPs during the development of morphine tolerance in an animal model of chronic inflammatory pain. Using behavioral approaches, we first established that repeated systemic morphine treatment induced morphine analgesic tolerance in rats coping with chronic inflammatory pain. We then observed that blockade of DOPs with subcutaneous naltrindole (NTI), a selective DOP antagonist, significantly attenuated the development of morphine tolerance in a dose‐dependent manner. We confirmed that this effect was DOP mediated by showing that an acute injection of NTI had no effect on morphine‐induced analgesia in naive animals. Previous pharmacological characterizations revealed the existence of DOP subtype 1 and DOP subtype 2. As opposed to NTI, 7‐benzylidenenaltrexone and naltriben were reported to be selective DOP subtype 1 and DOP subtype 2 antagonists, respectively. Interestingly, naltriben but not 7‐benzylidenenaltrexone was able to attenuate the development of morphine analgesic tolerance in inflamed rats. Altogether, our results suggest that targeting of DOP subtype 2 with antagonists provides a valuable strategy to attenuate the analgesic tolerance that develops after repeated morphine administration in the setting of chronic inflammatory pain.     

Comparison of flurbiprofen and alprazolam in the management of chronic pain syndrome

A three-process model of chronic pain comprising tissue damage, anxiety, and depression is hypothesized. Within this model, the effectiveness of flurbiprofen (for analgesia) plus either alprazolam (for anxiety and depression) or placebo was evaluated in a randomized, double-blind trial with a single crossover. Flurbiprofen was found to have a significant analgesic effect but this was not enhanced by combining it with alprazolam.     

Analgesic effects of intrathecally applied noradrenergic compounds in the developing rat: differences due to thermal vs mechanical nociception

Peak noradrenergic receptor development in rat spinal cord has been shown to occur around 12 days of postnatal life. The intent of the present study was to examine the development of analgesia produced by spinally applied noradrenergic agonists. The extent to which these drugs modulate pain information evoked by a thermal vs mechanical stimulus in the infant rat was also addressed. Intrathecal norepinephrine resulted in analgesia that was more pronounced against a mechanical than thermal stimulus and more pronounced in 10-day-olds than 3-day-olds. The alpha 2 receptor agonist clonidine produced a dose-dependent analgesia that first appeared at 7 days of age when tested with a thermal stimulus and 3 days of age when tested with a mechanical stimulus. The analgesic effect of clonidine was also greatest at 10 days of age. The alpha 1 agonist phenylephrine was without analgesic effects. The developmental profile of behavioral analgesia correlates with the ontogeny of noradrenergic receptor activity in the spinal cord. The finding that intrathecal norepinephrine produced a more pronounced analgesia against a mechanical rather than thermal stimulus in the adult is supported by our investigation in the infant rat.     

Corticotropin-releasing factor (CRF) produces analgesia in humans and rats

The analgesic activity of corticotropin releasing factor (CRF) was determined in a clinical model and in the rat hot plate test. Patients administered CRF reported significantly less postoperative pain than patients pretreated with placebo. In rats, injection of CRF resulted in a significant analgesia which was comparable in both intensity and duration to a 300 times greater molar dose of morphine. These findings suggest that endogenous CRF may play a physiologic role in modulating pain when released under conditions of stress.     

Correlation between the cumulative analgesic effect of electroacupuncture intervention and synaptic plasticity of hypothalamic paraventricular nucleus neurons in rats with sciatica

In the present study, a rat model of chronic neuropathic pain was established by ligation of the sciatic nerve and a model of learning and memory impairment was established by ovariectomy to investigate the analgesic effect of repeated electroacupuncture stimulation at bilateral Zusanli (ST36) and Yanglingquan (GB34). In addition, associated synaptic changes in neurons in the paraventricular nucleus of the hypothalamus were examined. Results indicate that the thermal pain threshold (paw withdrawal latency) was significantly increased in rats subjected to 2-week electroacupuncture intervention compared with 2-day electroacupuncture, but the analgesic effect was weakened remarkably in ovariectomized rats with chronic constrictive injury. 2-week electroacupuncture intervention substantially reversed the chronic constrictive injury-induced increase in the synaptic cleft width and thinning of the postsynaptic density. These findings indicate that repeated electroacupuncture at bilateral Zusanli and Yanglingquan has a cumulative analgesic effect and can effectively relieve chronic neuropathic pain by remodeling the synaptic structure of the hypothalamic paraventricular nucleus.     

Cholecystokinin‐8 antagonizes electroacupuncture analgesia through its B receptor in the caudate nucleus

Objectives: The analgesic effect of electroacupuncture (EA) stimulation has been proved. However, its mechanism of action is not clear. It has been well‐known that cholecystokinin‐8 (CCK‐8) is a neuropeptide which is mainly related to the mediation of pain. The caudate nucleus was selected to determine if the release of CCK and the neural activity in this nucleus were involved in producing EA analgesia. Materials and Methods: Radiant heat focused on the rat‐tail was used as the noxious stimulus. The pain threshold of rats was measured by tail‐flick latency (TFL). EA stimulation at the bilateral Zusanli (ST 36) acupoints of rats was used to investigate the effects of EA analgesia. The electrical activities of pain‐excited neurons (PEN) and pain‐inhibited neurons (PIN) in the caudate nucleus were recorded with a glass microelectrode. The present study examined the antagonistic effects of the intracerebral ventricular injection of CCK‐8 on EA analgesia and reversing effects of CCK‐B receptor antagonist (L‐365,260) injection into the caudate nucleus on CCK‐8. Results: The radiant heat focused on the tail of rats caused an increase in the evoked discharge of PEN and a reduction in the evoked discharge of PIN. EA stimulation at the bilateral ST 36 acupoints of rats resulted in the inhibition of PEN, the potentiation of PIN, and prolongation of TFL. The analgesic effect of EA was antagonized when CCK‐8 was injected into the intracerebral ventricle of rats. The antagonistic effect of CCK‐8 on EA analgesia was reversed by injection of CCK‐B receptor antagonist (L‐365,260) into the caudate nucleus of rats. Conclusions: Our results suggest that CCK‐8 antagonize EA analgesia through its B receptor.     

Analgesia for tonic pain by self-administered lateral hypothalamic stimulation.

The present study was designed to determine if analgesia for tonic pain could be induced with self-administered lateral hypothalamic (LH) stimulation. The majority of studies of LH stimulation employed behaviorally non-contingent stimulation. According to some investigators behaviorally non-contingent LH stimulation might be aversive and consequently yield stress-induced analgesia rather than a primary analgesic effect. Other investigators have reported that non-contingent LH stimulation can be rewarding. Our findings indicate that LH analgesia for tonic pain can be obtained with self-administered LH stimulation. These findings indicate that the analgesia we have obtained in previous work is a primary effect and not dependent on stress induced by behaviorally non-contingent LH stimulation.     

High-frequency rTMS over the dorsolateral prefrontal cortex on chronic and provoked pain: A systematic review and meta-analysis

BackgroundHigh-frequency rTMS over the dorsolateral prefrontal cortex (DLPFC) has demonstrated mixed effects on chronic and provoked pain.Objectives/MethodsIn this study, a meta-analysis was conducted to characterise the potential analgesic effects of high-frequency rTMS over the DLPFC on both chronic and provoked pain.ResultsA total of 626 studies were identified in a systematic search. Twenty-six eligible studies were included for the quantitative review, among which 17 modulated chronic pain and the remaining investigated the influence on provoked pain. The left side DLPFC was uniformly targeted in the chronic pain studies. While our data identified no overall effect of TMS across chronic pain conditions, there was a significant short-term analgesia in neuropathic pain conditions only (SMD = −0.87). In terms of long-lasting analgesia, there was an overall pain reduction in the midterm (SMD = −0.53, 24.6 days average) and long term (SMD = −0.63, 3 months average) post DLPFC stimulation, although these effects were not observed within specific chronic pain conditions. Surprisingly, the number of sessions was demonstrated to have no impact on rTMS analgesia. In the analysis of provoked pain, our data also indicated a significant analgesic effect following HF-rTMS over the DLPFC (SMD = −0.73). Importantly, we identified a publication bias in the studies of provoked pain but not for chronic pain conditions.ConclusionsOverall, our findings support that HF-DLPFC stimulation is able to induce an analgesic effect in chronic pain and in response to provoked pain. These results highlight the potential of DLPFC-rTMS in the management of certain chronic pain conditions and future directions are discussed to enhance the potential long-term analgesic effects.     

Rats with decreased brain cholecystokinin levels show increased responsiveness to peripheral electrical stimulation-induced analgesia

Using the P77PMC strain of rat, which is genetically prone to audiogenic seizures, and also has decreased levels of cholecystokinin (CCK), we examined the analgesic response to peripheral electrical stimulation, which is, in part, opiate-mediated. A number of studies have suggested that CCK may function as an antagonist to endogenous opiate effects. Therefore, we hypothesized that the P77PMC animals would show an enhanced analgesic response based on their decreased CCK levels producing a diminished endogenous opiate antagonism. We found that the analgesic effect on tail flick latency produced by 100 Hz peripheral electrical stimulation was more potent and longer lasting in P77PMC rats than in control rats. Moreover, the potency of the stimulation-produced analgesia correlated with the vulnerability to audiogenic seizures in these rats. We were able to block the peripheral electrical stimulation-induced analgesia (PSIA) using a cholecystokinin octapeptide (CCK-8) administered parenterally. Radioimmunoassay showed that the content of CCK-8 in cerebral cortex, hippocampus and periaqueductal gray was much lower in P77PMC rat than in controls. These results suggest that low CCK-8 content in the central nervous system of the P77PMC rats may be related to the high analgesic response to peripheral electrical stimulation, and further support the notion that CCK may be an endogenous opiate antagonist.     

The effect of pituitary removal on pain regulation in the rat

The effects of hypophysectomy (HX) on pain regulation in basal and in various stressful situations were investigated in the rat. Pain sensitivity was assessed by measuring the thresholds of 3 nociceptive reactions (tail withdrawal, vocalization, vocalization afterdischarge) following electrical stimulation of the tail. The completeness of HX and the integrity of hypothalamus were verified in each HX rat.(1) Baseline pain thresholds were lower in HX rats than in shan-operated animals; (2) naloxone (Nx) hyperalgesia was only slightly altered by HX; (3) different types of stress induced different types of changes in nociception i.e. analgesia or hyperalgesia. The influence of HX varied according to the stress: it increased hyperalgesia, reduced analgesia, or had no effect at all.These results indicate that in the rat: (i) the pituitary participates in the regulation of basal pain sensitivity, probably through analgesic factors; (ii) Nx hyperalgesia results essentially from an antagonism of endogenous opioids originating in the CNS and not in the pituitary; and (iii) the pain regulatory processes engaged in adaptation to stressful stimuli involve the CNS and the pituitary in variable proportions depending upon the nature of the stress.     

Thalamocortical Mechanisms for Nostalgia-Induced Analgesia

As a predominately positive emotion, nostalgia serves various adaptive functions, including a recently revealed analgesic effect. The current fMRI study aimed to explore the neural mechanisms underlying the nostalgia-induced analgesic effect on noxious thermal stimuli of different intensities. Human participants'' (males and females) behavior results showed that the nostalgia paradigm significantly reduced participants'' perception of pain, particularly at low pain intensities. fMRI analysis revealed that analgesia was related to decreased brain activity in pain-related brain regions, including the lingual and parahippocampal gyrus. Notably, anterior thalamic activation during the nostalgia stage predicted posterior parietal thalamus activation during the pain stage, suggesting that the thalamus might play a key role as a central functional linkage in the analgesic effect. Moreover, while thalamus-PAG functional connectivity was found to be related to nostalgic strength, periaqueductal gray-dorsolateral prefrontal cortex (PAG-dlPFC) functional connectivity was found to be associated with pain perception, suggesting possible analgesic modulatory pathways. These findings demonstrate the analgesic effect of nostalgia and, more importantly, shed light on its neural mechanism.SIGNIFICANCE STATEMENT Nostalgia is known to reduce individuals'' perception of physical pain. The underlying brain mechanisms, however, are unclear. Our study found that the thalamus plays a key role as a functional linkage between nostalgia and pain, suggesting a possible analgesic modulatory mechanism of nostalgia. These findings have implications for the underlying brain mechanisms of psychological analgesia.