R. Sun

The role of calcitonin gene-related peptide (CGRP) in the generation and maintenance of mechanical allodynia and hyperalgesia in rats after intradermal injection of capsaicin.

&NA; This study was designed to assess the role of calcitonin gene‐related peptide (CGRP) and its receptor in the generation and maintenance of secondary mechanical allodynia and hyperalgesia induced by intradermal injection of capsaicin in rats. Paw withdrawal responses (PWRs) to von Frey hairs with different bending forces applied on the rat paw were tested in this study. CGRP8–37, a specific antagonist of CGRP 1 receptors, was delivered through a microdialysis fiber inserted across the dorsal horn. Post‐ and pretreatment paradigms were followed. When CGRP8–37 was administered 1 h after capsaicin injection, the mechanical allodynia and hyperalgesia were partially reversed in a dose‐dependent manner. On the other hand, when rats were treated with CGRP8–37 prior to capsaicin injection, the PWRs to von Frey applications were significantly reduced as compared to control animals. Collectively, these results suggest that CGRP receptors present in the dorsal horn are involved in the generation and maintenance of nociceptive behaviors associated with cutaneous inflammation.

Activation of protein kinase B/Akt in the periphery contributes to pain behavior induced by capsaicin in rats

Protein kinase B (PKB/Akt) is a member of the second-messenger regulated subfamily of protein kinases. It is implicated in signaling downstream of growth factors, insulin receptor tyrosine kinases and phosphoinositide 3-kinase (PI3K). Current studies indicate that nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and PI3K help mediate inflammatory hyperalgesia. However, little is known about the role of PKB/Akt in the nociceptive system. In this study, we investigated whether PKB/Akt in primary sensory neurons is activated after noxious stimulation and contributes to pain behavior induced in rats by capsaicin. We demonstrated that phospho-PKB/Akt (p-PKB/Akt) is increased in dorsal root ganglia (DRG) at 5 min after intradermal injection of capsaicin. p-PKB/Akt is distributed predominantly in small- and medium-sized DRG cells. After capsaicin injection, p-PKB/Akt (473) is colocalized with isotectin-B4 (IB4), tyrosine kinase A (TrkA), and calcitonin gene-related peptide (CGRP). Furthermore, most transient receptor potential vanilloid type 1 (TRPV1) positive DRG neurons double label for p-PKB/Akt. Behavioral experiments show that intradermal injection of a PI3K (upstream of PKB/Akt) inhibitor, wortmannin, dose-dependently inhibits the changes in exploratory behavior evoked by capsaicin injection. The PKB/Akt inhibitor, Akt inhibitor IV, has the same effect. The results suggest that the PKB/Akt signaling pathway in the periphery is activated by noxious stimulation and contributes to pain behavior.

Activation of protein kinase B/Akt signaling pathway contributes to mechanical hypersensitivity induced by capsaicin.

Abstract We investigated the involvement of the protein kinase B/Akt (PKB/Akt) signaling pathway in the mechanical hypersensitivity induced in rats by capsaicin. Intradermal injection of capsaicin results in activation of PKB/Akt in the lumbar spinal cord, most prominently in the dorsal horn, starting by 5 min after capsaicin injection and lasting at least 1 h. The activated PKB/Akt in the spinal cord is in neurons, since phospho‐PKB/Akt (p‐PKB/Akt) colocalizes with the neuronal marker, neuronal‐specific nuclear protein (NeuN). The mechanical hypersensitivity is shown by the enhanced paw withdrawal frequency to applications of von Frey filaments with different bending forces (30, 100, 200 mN) on the rat paw. Pre‐treatment with several different PKB/Akt inhibitors, including SH‐6, Akt inhibitor IV, and Akt inhibitor V, blocked the mechanical hypersensitivity induced by intradermal injection of capsaicin, a measure of spinal cord central sensitization. Two structurally unrelated phosphoinositide 3‐Kinase (PI3K, upstream of PKB/Akt) inhibitors, Wortmannin and LY294002, also prevented the mechanical hypersensitivity induced by intradermal injection of capsaicin. Furthermore, post‐treatment with the PI3K inhibitor, Wortmannin, or PKB/Akt inhibitors, such as NL‐71‐101, SH‐6, Akt inhibitor IV, and inhibitor V significantly reduced the established mechanical hypersensitivity induced by capsaicin. The PKB/Akt signaling pathway in the spinal cord is therefore involved in pain hypersensitivity.

Effects of intrathecal injections of melatonin analogs on capsaicin-induced secondary mechanical allodynia and hyperalgesia in rats.

&NA; Melatonin, its agonists/antagonists were administered intrathecally (i.t.) before/after intradermal injection of capsaicin. Capsaicin produced an increase in the paw withdrawal frequency (PWF) in the presumed area of secondary mechanical allodynia and hyperalgesia. Melatonin agonists in the absence of a capsaicin injection decreased the PWF significantly, whereas melatonin antagonists given intrathecally alone were ineffective in the absence of a capsaicin injection. Pre‐treatment with a melatonin agonist i.t. caused a reduction in the PWF after capsaicin. In contrast, the PWF increased after capsaicin with pre‐administration of a melatonin antagonist i.t. Combined pre‐treatment with melatonin and a melatonin antagonist i.t. prevented the change in PWF induced by melatonin alone after capsaicin. Intrathecal post‐treatment with a melatonin agonist reduced the enhanced PWF that followed an injection of capsaicin, but treatment with a combination of a melatonin agonist and its antagonist did not alter the responses. The PWF was unaffected when melatonin analogs were applied i.t. at the T6 level or were injected intramuscularly adjacent to the L4 vertebra. In spinal rats, the data showed comparable effects of melatonin analogs on capsaicin‐induced secondary mechanical hyperalgesia. Animal motor function tested by ‘activity box’ showed that motion activity was not affected by i.t. melatonin or its antagonist. These results suggest that activation of the endogenous melatonin system in the spinal cord can reduce the generation, development and maintenance of central sensitization, with a resultant inhibition of capsaicin‐induced secondary mechanical allodynia and hyperalgesia.

Intradermal injection of capsaicin induces acute substance P release from rat spinal cord dorsal horn.

Increased release of substance P (SP) from the dorsal horn following noxious stimuli, such as spinal administration of capsaicin, has been demonstrated in previous studies. However, changes in the release of SP in response to intradermal injection of capsaicin still remain unknown. This study was designed to demonstrate in vivo spinal SP release following intradermal injection of capsaicin (3%, 50 microl), using polyimide tubing with a single hole introduced into the rat dorsal horn. The changes in the content of SP in the rat dorsal horn tissues before and after capsaicin (3%, 50 microl) injection were also investigated. The SP concentration in the samples was analyzed using an enzyme-linked immunosorbent assay (ELISA). We found that intradermal injection of capsaicin induced a quick SP release within the dorsal horn. The peak of the release appeared around 10 min after the injection. In contrast, intradermal injection of capsaicin had no significant effect on the SP content in the dorsal horn. This study has provided direct evidence of the effect of intradermal injection of capsaicin on SP release within the dorsal horn, with the major source being from the central terminals of primary afferents.