Differential Roles of Spinal Protein Kinases C and A in Development of Primary Heat and Mechanical Hypersensitivity Induced by Subcutaneous Bee Venom Chemical Injury in the Rat

Abstract

It has been demonstrated that subcutaneous injection of bee venom (BV) can produce different types of pain and hypersensitivity including persistent spontaneous nociception (PSN), primary heat and mechanical hypersensitivity (hyperalgesia) and mirror-image heat (MIH) hypersensitivity in an individual animal, and the changes of spinal neurons are likely to be responsible for the production of these pain-related behaviors. In this study, we examined the roles of spinal protein kinase C (PKC) and protein kinase A (PKA) in the BV-induced different types of pain and hypersensitivity in conscious rats. We found that: (1) BV-induced primary heat hypersensitivity could be blocked by intrathecal pre- or posttreatment with a PKC inhibitor, chelerythrine chloride (CH), while a PKA inhibitor, N-(2-[p-bromocinnamylamino]ethyl)-5-isoquinolinesulfonamide hydrochloride (H89), had no effect. (2) BV-induced primary mechanical hypersensitivity could be blocked by pre- or posttreatment with H89, whereas CH had no effect. (3) Both pre- and posttreatment with H89 produced suppressive effects on both induction and maintenance of the BV-induced PSN and MIH hypersensitivity. Based on the present findings, we proposed that spinal PKC might be activated during the central processes of primary heat hypersensitivity, while spinal PKA is likely to be involved in primary mechanical hypersensitivity induced by subcutaneous BV chemical injury. Taken together with our previous report however, spinal PKC and PKA are likely to be simultaneously involved in the central processes of both PSN and MIH hypersensitivity.

The nociceptive and anti-nociceptive effects of bee venom injection and therapy: A double-edged sword

“…administration of chelerythrine chloride (CH), a PKC inhibitor and H89 (N-(2-[ p -bromocinnamylamino]ethyl)-5-isoquinolinesulfonamide hydrochloride), a PKA inhibitor (Li et al, 2000b; Li and Chen, 2003). SQ22536 was also administered to inhibit adenylate cyclase (AC), which produces cAMP that is required by PKA to function (Li and Chen, 2003).…”

“…administration of chelerythrine chloride (CH), a PKC inhibitor and H89 (N-(2-[ p -bromocinnamylamino]ethyl)-5-isoquinolinesulfonamide hydrochloride), a PKA inhibitor (Li et al, 2000b; Li and Chen, 2003). SQ22536 was also administered to inhibit adenylate cyclase (AC), which produces cAMP that is required by PKA to function (Li and Chen, 2003). For the bee venom-induced persistent nociception, pre-treatment of spinal cord with three drugs of CH, H89 or SQ22536 at three separate doses significantly prevented the paw flinches from occurring in a dose-related manner, while only CH and H89 produced effective suppression following post-treatment of the highest dose used (Li et al, 2000b; Li and Chen, 2003).…”

The Many Faces of H89: A Review

“…Determining the effects of kinase inhibitors on protein substrates in vivo is, however, of central importance relative to inferences to inhibitor specificity and its effects at organ level. A few short term studies lasting 5-120 min using intrathecal, topical, subcutaneous or intracerebroventricular administration of H89 have been reported (20,50,51). A recent study (18) applied differential proteomic methods to measure protein phosphorylation in vivo.…”

“…In summary, several PKA inhibitors are currently available, with different properties and IC 50 values, which should be taken into account when designing an experiment. Both Rp-cAMPS and synthetic PKI analogs appear to be more specific inhibitors of PKA than H89.…”

Proteomic Analysis of Rat Liver Phosphoproteins after Treatment with Protein Kinase Inhibitor H89 (N-(2-[p-Bromocinnamylamino-]ethyl)-5-isoquinolinesulfonamide)

“…H89 has been widely used as a tool to inhibit PKA in cell-based or biochemical assays; however, mammalian studies using H89 in vivo are limited. Short-term studies that used intrathecal, topical, s.c., or intracerebroventricular administration have been reported with durations of treatment ranging from 5 to 120 min (Hua et al, 1999;Dolan and Nolan, 2001;Vargas et al, 2001;Fang et al, 2003;Li and Chen, 2003;Sun et al, 2004;Lim et al, 2005). Recently, H89 administration to isolated rat hearts has been reported (Makaula et al, 2005).…”