Korean J Pain.  2020 Jan;33(1):30-39. 10.3344/kjp.2020.33.1.30.

Gabexate mesilate ameliorates the neuropathic pain in a rat model by inhibition of proinflammatory cytokines and nitric oxide pathway via suppression of nuclear factor-κB

• Oh SH
• Lee HY
• Ki YJ
• Kim SH
• Lim KJ
• Jung KT

• KMID: 2506640
• DOI: http://doi.org/10.3344/kjp.2020.33.1.30

Abstract

Background
This study examined the effects of gabexate mesilate on spinal nerve ligation (SNL)-induced neuropathic pain. To confirm the involvement of gabexate mesilate on neuroinflammation, we focused on the activation of nuclear factor-κB (NF-κB) and consequent the expression of proinflammatory cytokines and inducible nitric oxide synthase (iNOS).
Methods
Male Sprague-Dawley rats were used for the study. After randomization into three groups: the sham-operation group, vehicle-treated group (administered normal saline as a control), and the gabexate group (administered gabexate mesilate 20 mg/kg), SNL was performed. At the 3rd day, mechanical allodynia was confirmed using von Frey filaments, and drugs were administered intraperitoneally daily according to the group. The paw withdrawal threshold (PWT) was examined on the 3rd, 7th, and 14th day. The expressions of p65 subunit of NF-κB, interleukin (IL)-1, IL-6, tumor necrosis factor-α, and iNOS were evaluated on the 7th and 14th day following SNL.
Results
The PWT was significantly higher in the gabexate group compared with the vehicle-treated group (p < 0.05). The expressions of p65, proinflammatory cyto kines, and iNOS significantly decreased in the gabexate group compared with the vehicle-treated group (p < 0.05) on the 7th day. On the 14th day, the expressions of p65 and iNOS showed lower levels, but those of the proinflammatory cytokines showed no significant differences.
Conclusions
Gabexate mesilate increased PWT after SNL and attenuate the pro gress of mechanical allodynia. These results seem to be involved with the antiinflammatory effect of gabexate mesilate via inhibition of NF-κB, proinflammatory cytokines, and nitric oxide.

Keyword

Analgesia; Gabexate; Hyperalgesia; Human; Inflammation; Neuralgia; NF-Kappa B; Nitric Oxide Synthase Type II; Spinal Nerves

Figure

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  Fig. 1 CONSORT diagram of the study design.

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  Fig. 2 Paw withdrawal threshold (PWT) across time. The PWT was observed from the 3rd day until the 14th day following spinal nerve ligation (SNL). On the 3rd day after SNL, mechanical allodynia was confirmed. The rats of the gabexate group showed higher PWT than those of the vehicle-treated group during the observation period (P < 0.001). The sham-operation group, sham operation only without SNL or drug; the vehicle-treated group, the control group treated with intraperitoneal normal saline daily after 3rd day following SNL until the end of the study period; the gabexate group, the experimental group treated with intraperitoneal 20 mg/kg of gabexate mesilate daily after 3rd day following SNL until the end of the study period. *P < 0.05 compared with the vehicle-treated group.

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  Fig. 3 The expression of p65 subunit of nuclear factor-κB at the ipsilateral dorsal horn of L5 spinal cord across time. The levels of p65 were significantly increased in the vehicle-treated group and gabexate group compared with the sham-operation group (P < 0.001). The expression of p65 was markedly decreased in the gabexate group compared with the vehicle-treated group (P < 0.05) throughout the experimental period. The sham-operation group, sham operation only without spinal nerve ligation (SNL) or drug; the vehicle-treated group, the control group treated with intraperitoneal normal saline daily after 3rd day following SNL until the end of the study period; the gabexate group, the experimental group treated with intraperitoneal 20 mg/kg of gabexate mesilate daily after 3rd day following SNL until the end of the study period.

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  Fig. 4 The levels of interleukin (IL)-1 (A), IL-6 (B), and tumor necrosis factor-alpha (TNF-α; C) at the ipsilateral dorsal horn of L5 spinal cord across time. The levels IL-1, IL-6, and TNF-α were measured at the 7th and the 14th day following spinal nerve ligation (SNL). The expressions of IL-1, IL-6, and TNF-α were significantly increased in the vehicle-treated group and the gabexate group compared with the sham-operation group at both 7th and 14th day after SNL (P < 0.05). After administration of gabexate mesilate, the expressions of IL-1, IL-6, and TNF-α were significantly attenuated in the gabexate group compared with the vehicle-treated group at 7th day after SNL (P < 0.05). However, there were no statistically significances in the expressions of IL-1, IL-6, and TNF-α between the vehicle-treated group and the gabexate group at 14th day (P > 0.05). The sham-operation group, sham group; the vehicle-treated group, the control group treated with intraperitoneal normal saline daily after 3rd day following SNL until the end of the study period; Group E, the experimental group treated with intraperitoneal 20 mg/kg of gabexate mesilate daily after 3rd day following SNL until the end of the study period.

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  Fig. 5 The expression of inducible nitric oxide synthase (iNOS) at the ipsilateral dorsal horn of L5 spinal cord across time. The levels of iNOS were significantly increased in the vehicle-treated group and the experimental group (group E) compared with the sham-operation group (P < 0.05). The expression of iNOS was markedly decreased in group E compared with the vehicle-treated group throughout the experimental period (P < 0.01). The sham-operation group, sham group; the vehicle-treated group, the control group treated with intraperitoneal normal saline daily after 3rd day following spinal nerve ligation (SNL) until the end of the study period; Group E treated with intraperitoneal 20 mg/kg of gabexate mesilate daily after 3rd day following SNL until the end of the study period.

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