6'-Guanidinonaltrindole (6'-GNTI) Is a G Protein-biased κ-Opioid Receptor Agonist That Inhibits Arrestin Recruitment

κ-Opioid receptor (KOR) agonists do not activate the reward pathway stimulated by morphine-like μ-opioid receptor (MOR) agonists and thus have been considered to be promising nonaddictive analgesics. However, KOR agonists produce other adverse effects, including dysphoria, diuresis, and constipation. The therapeutic promise of KOR agonists has nonetheless recently been revived by studies showing that their dysphoric effects require arrestin recruitment, whereas their analgesic effects do not. Moreover, KOR agonist-induced antinociceptive tolerance observed in vivo has also been proposed to be correlated to the ability to induce arrestin-dependent phosphorylation, desensitization, and internalization of the receptor. The discovery of functionally selective drugs that are therapeutically effective without the adverse effects triggered by the arrestin pathway is thus an important goal. We have identified such an extreme G protein-biased KOR compound, 6'-guanidinonaltrindole (6'-GNTI), a potent partial agonist at the KOR receptor for the G protein activation pathway that does not recruit arrestin. Indeed, 6'-GNTI functions as an antagonist to block the arrestin recruitment and KOR internalization induced by other nonbiased agonists. As an extremely G protein-biased KOR agonist, 6'-GNTI represents a promising lead compound in the search for nonaddictive opioid analgesic as its signaling profile suggests that it will be without the dysphoria and other adverse effects promoted by arrestin recruitment and its downstream signaling.     

Crypteins derived from the mouse neuropeptide FF (NPFF)A precursor display NPFF-like effects in nociceptive tests in mice

NPFF precursor, pro-NPFF(A) contains three known bioactive sequences: NPFF (FLFQPQRF-NH(2)), neuropeptide AF (NPAF; AGEGLSSPFWSLAAPQRF-NH(2)) and neuropeptide SF (NPSF; SLAAPQRF-NH(2)). The key-feature of these fragments is their common PQRF-amidated sequence at their C termini. Here, we evaluated the biological activity of two other sequences derived from the mouse NPFF(A) precursor, that does not have PQRF-amidated C-terminus. One peptide was residing between positions 85 and 99 in the mice pro-NPFF(A). This peptide was referred to as neuropeptide SA (NPSA; SAWGSWSKEQLNPQA), assigned due to its flanking amino acids. Another sequence used in the experiments was N-terminal fragment of NPSA, here referred to as neuropeptide SS (NPSS; SAWGSWS). These two peptides, classified as crypteins, were synthesized and tested in the hot-plate and tail immersion tests in mice for their pharmacological activity in morphine-induced antinociception. The effects of both crypteins were compared to NPFF. Our experiments indicated that both crypteins inhibited morphine antinociception and their effects were reversed by RF9, an antagonist of NPFF receptors. These data show that NPSA and NPSS possess NPFF-like anti-opioid activity in these behavioral tests.     

Involvement of ATP in noxious stimulus-evoked release of glutamate in rat medullary dorsal horn: A microdialysis study

Our electrophysiological studies have shown that both purinergic and glutamatergic receptors are involved in central sensitization of nociceptive neurons in the medullary dorsal horn (MDH). Here we assessed the effects of intrathecal administration of apyrase (a nucleotide degrading enzyme of endogenous adenosine 5-triphosphate [ATP]), a combination of apyrase and 1,3-dipropyl-8-cyclopentylxanthine (DPCPX, an adenosine A1 receptor antagonist), or 2,3-O-2,4,6-trinitrophenyl-adenosine triphosphate (TNP-ATP, a P2X1, P2X3, P2X2/3 receptor antagonist) on the release of glutamate in the rat MDH evoked by application of mustard oil (MO) to the molar tooth pulp. In vivo microdialysis was used to dialyse the MDH every 5 min, and included 3 basal samples, 6 samples after drug treatment and 12 samples following application of MO. Tooth pulp application of MO induced a significant increase in glutamate release in the MDH. Superfusion of apyrase or TNP-ATP alone significantly reduced the MO-induced glutamate release in the MDH, as compared to vehicle. Furthermore, the suppressive effects of apyrase on glutamate release were reduced by combining it with DPCPX. This study demonstrates that application of an inflammatory irritant to the tooth pulp induces glutamate release in the rat MDH in vivo that may be reduced by processes involving endogenous ATP and adenosine.     

Local ASIC3 modulates pain and disease progression in a rat model of osteoarthritis

Background

Recent data have suggested a relationship between acute arthritic pain and acid sensing ion channel 3 (ASIC3) on primary afferent fibers innervating joints. The purpose of this study was to clarify the role of ASIC3 in a rat model of osteoarthritis (OA) which is considered a degenerative rather than an inflammatory disease.

Methods

We induced OA via intra-articular mono-iodoacetate (MIA) injection, and evaluated pain-related behaviors including weight bearing measured with an incapacitance tester and paw withdrawal threshold in a von Frey hair test, histology of affected knee joint, and immunohistochemistry of knee joint afferents. We also assessed the effect of ASIC3 selective peptide blocker (APETx2) on pain behavior, disease progression, and ASIC3 expression in knee joint afferents.

Results

OA rats showed not only weight-bearing pain but also mechanical hyperalgesia outside the knee joint (secondary hyperalgesia). ASIC3 expression in knee joint afferents was significantly upregulated approximately twofold at Day 14. Continuous intra-articular injections of APETx2 inhibited weight distribution asymmetry and secondary hyperalgesia by attenuating ASIC3 upregulation in knee joint afferents. Histology of ipsilateral knee joint showed APETx2 worked chondroprotectively if administered in the early, but not late phase.

Conclusions

Local ASIC3 immunoreactive nerve is strongly associated with weight-bearing pain and secondary hyperalgesia in MIA-induced OA model. APETx2 inhibited ASIC3 upregulation in knee joint afferents regardless of the time-point of administration. Furthermore, early administration of APETx2 prevented cartilage damage. APETx2 is a novel, promising drug for OA by relieving pain and inhibiting disease progression.     

The role of activin in neuropeptide induction and pain sensation

Signals from target tissues play critical roles in the functional differentiation of neuronal cells, and in their subsequent adaptations to peripheral changes in the adult. Sensory neurons in the dorsal root ganglia (DRG) provide an excellent model system for the study of signals that regulate the development of neuronal diversity. DRG have been well characterized and contain both neurons that convey information from muscles about limb position, as well as other neurons that provide sensations from skin about pain information. Sensory neurons involved in pain sensation can be distinguished physiologically and antigenically, and one hallmark characteristic is that these neurons contain neuropeptides important for their functions. The transforming growth factor (TGF) beta family member activin A has recently been implicated in neural development and response to injury. During sensory neuron development, peripheral target tissues containing activin or activin itself can regulate pain neuropeptide expression. Long after development has ceased, skin target tissues retain the capacity to signal neurons about changes or injury, to functionally refine synapses. This review focuses on the role of activin as a target-derived differentiative factor in neural development that has additional roles in response to cutaneous injuries in the adult.     

Selective P2X7 receptor antagonists for chronic inflammation and pain

ATP, acting on P2X₇ receptors, stimulates changes in intracellular calcium concentrations, maturation, and release of interleukin-1β (IL-1β), and following prolonged agonist exposure, cell death. The functional effects of P2X₇ receptor activation facilitate several proinflammatory processes associated with arthritis. Within the nervous system, these proinflammatory processes may also contribute to the development and maintenance of chronic pain. Emerging data from genetic knockout studies have indicated specific roles for P2X₇ receptors in inflammatory and neuropathic pain states. The discovery of multiple distinct chemical series of potent and highly selective P2X₇ receptor antagonists have enhanced our understanding of P2X₇ receptor pharmacology and the diverse array of P2X₇ receptor signaling mechanisms. These antagonists have provided mechanistic insight into the role(s) P2X₇ receptors play under pathophysiological conditions. In this review, we integrate the recent discoveries of novel P2X₇ receptor-selective antagonists with a brief update on P2X₇ receptor pharmacology and its therapeutic potential.     

The role of excitatory amino acid receptors and intracellular messengers in persistent nociception after tissue injury in rats

Increased pain sensitivity (hyperalgesia) and persistent nociception following peripheral tissue injury depends both on an increase in the sensitivity of primary afferent nociceptors at the site of injury (peripheral sensitization), and on an increase in the excitability of neurons in the central nervous system (central sensitization). We will review evidence that central sensitization, and the persistent nociception it leads to, are dependent on an action of glutamate and aspartate at excitatory amino acid (EAA) receptors. Additional evidence will be presented implicating a role of various intracellular second messengers that are coupled to EAA receptors (nitric oxide, arachidonic acid, and protein kinase C) to central sensitization and persistent nociception following tissue injury. Finally, we will examine the evidence for a contribution of molecular events, including noxious stimulus-induced expression of immediate-early genes such as c-fos to persistent nociception.     

腰痹通胶囊联合塞来昔布胶囊治疗椎间盘源性腰痛的疗效及对血清炎性因子和疼痛介质的影响

摘要 目的：探讨腰痹通胶囊联合塞来昔布胶囊治疗椎间盘源性腰痛（DLBP）的疗效及对血清炎性因子和疼痛介质的影响。方法：选择2017年5月~2022年5月期间我院接收的75例DLBP患者。依据信封抽签法将患者分为对照组（n=37）和研究组（n=38）,对照组接受塞来昔布胶囊治疗,研究组接受腰痹通胶囊联合塞来昔布胶囊治疗。对比两组疗效、临床症状评分、血清炎性因子和疼痛介质的变化情况,同时观察两组用药安全性。结果：研究组痊愈8例,显效10例,有效18例,临床总有效率94.74%,高于对照组的痊愈4例,显效8例,有效17例,临床总有效率78.38%（P<0.05）。治疗1个月后,研究组的视觉疼痛模拟（VAS）评分、Oswestry功能障碍指数（ODI）较对照组更低（P<0.05）,改良日本骨科协会（JOA）评分较对照组更高（P<0.05）。治疗1个月后,研究组的血清白细胞介素-6（IL-6）、C反应蛋白（CRP）、肿瘤坏死因子-α（TNF-α）水平较对照组更低（P<0.05）。治疗1个月后,研究组的血清 P物质（SP）、多巴胺（DA）、前列腺素E₂（PGE₂）水平较对照组更低（P<0.05）。两组不良反应发生率组间对比差异不显著（P>0.05）。结论：腰痹通胶囊联合塞来昔布胶囊治疗DLBP,可有效减轻疼痛,恢复腰椎功能,降低血清炎性因子和疼痛介质水平。     

苦参碱对神经病理性大鼠背根神经节P2X3受体、疼痛行为学和疼痛阈值的影响

摘要 目的：探讨苦参碱对神经病理性大鼠背根神经节P2X3受体、疼痛行为学和疼痛阈值的影响。方法：选择Sprague-Dawley雄性大鼠30只,随机分为3组,包括模型组、试验组和假手术组。于大鼠造模成功1 d后,试验组给予30 mg/（kgod）的剂量在腹腔注射苦参碱溶液,1次/d;给予假手术组和模型组腹腔注射等量浓度为0.9 %的氯化钠溶液,1次/d,共14 d。进行自发疼痛行为学评分检测、机械痛阈值检测、热痛阈值检测、P2X2和P2X3mRNA相对表达量检测、P2X2和P2X3蛋白表达水平检测,以及氧化应激指标水平检测。结果：术后模型组与试验组自发性疼痛行为学评分与假手术组比均升高,自术后第5天起,与模型组比,试验组自发性疼痛行为学评分明显低于模型组（P<0.05）;自术后第3天起,相较于假手术组,模型组机械痛阈值、热痛阈值显著下降,相较于模型组,试验组自术后第5天起机械痛阈值、热痛阈值显著上升（均P<0.05）;术后第14天试验组与假手术组机械痛阈值、热痛阈值对比无差异（P>0.05）;模型组P2X2和P2X3mRNA、P2X2及P2X3蛋白比假手术组和试验组高（均P<0.05）,试验组和假手术组P2X2、P2X3mRNA、P2X2及P2X3蛋白比较无差异（P>0.05）;干预前及干预1、2周后模型组大鼠脊髓组织SOD比假手术组低,MDA比假手术组高;试验组大鼠脊髓组织SOD比模型组高,MDA比模型组低（均P<0.05）。结论：苦参碱可有效缓解神经病理性痛的所引发的机械痛觉和热痛觉,镇痛作用较好,机制可能在于其可使大鼠背根神经元中P2X2、P2X3受体下降相关,同时其在抑制神经病理性大鼠脊髓组织氧化应激反应方面有一定的作用,与其在对神经病理性痛大鼠脊髓组织神经元凋亡的抑制有密切关系。     

Mechanisms of analgesic effect of mesenchymal stem cells in osteoarthritis pain

Osteoarthritis (OA) is the most common musculoskeletal disease, and it is a major cause of pain, disability and health burden. Pain is the most common and bothersome presentation of OA, but its treatment is still suboptimal, due to the short-term action of employed analgesics and their poor adverse effect profile. Due to their regenerative and anti-inflammatory properties, mesenchymal stem cells (MSCs) have been extensively investigated as a potential therapy for OA, and numerous preclinical and clinical studies found a significant improvement in joint pathology and function, pain scores and/or quality of life after administration of MSCs. Only a limited number of studies, however, addressed pain control as the primary end-point or investigated the potential mechanisms of analgesia induced by MSCs. In this paper, we review the evidence reported in literature that support the analgesic action of MSCs in OA, and we summarize the potential mechanisms of these antinociceptive effects.