J Korean Neurosurg Soc.  2016 Mar;59(2):98-105. 10.3340/jkns.2016.59.2.98.

The Effect of GCSB-5 a New Herbal Medicine on Changes in Pain Behavior and Neuroglial Activation in a Rat Model of Lumbar Disc Herniation

• Cho HK
• Kim SY
• Choi MJ
• Baek SO
• Kwak SG
• Ahn SH

• KMID: 2192038
• DOI: http://doi.org/10.3340/jkns.2016.59.2.98

Abstract

OBJECTIVE
Lumbar disc herniation can induce sciatica by mechanical compression and/or chemical irritation. The aim of this study was to compare the effects of GCSB-5 (Shinbaro(R)) and NSAIDs on pain-related behavior and on the expressions of microglia, astrocytes, CGRP, TRPV1, IL-6, and CX3CL1 in a rat model of lumbar disc herniation.
METHODS
112 male Sprague-Dawley rats underwent implantation of nucleus pulposus to a dorsal root ganglion (DRG). Rats were divided into five groups as follows; a saline group (the vehicle control group) (n=27), a 10 mg/kg aceclofenac group (the aceclofenac group) (n=22), and 100, 300 or 600 mg/kg GCSB-5 groups (the GCSB-5 100, 300, or 600 groups) (n=21 for each group). Rats were tested for mechanical allodynia at 3 days after surgery and at 1 day, 3 days, 7 days, 14 days, 21 days, 28 days, 35 days, 42 days, 49 days, and 56 days after treatment commencement. Immunohistochemical staining of microglia (Iba1), astrocytes (GFAP), CGRP, and TRPV1, and PCR for IL-6 and CX3CL1 were performed on spinal dorsal horns and DRGs at 56 days after medication commencement.
RESULTS
After 56 days of GCSB-5 300 administration, mechanical withdrawal thresholds were significantly increased (p<0.05), and immunohisto-chemical expressions of Iba1, GFAP, CGRP, and TRPV1 were reduced than other groups, but this difference was not statistically significant.
CONCLUSION
These results indicate GCSB-5 reduces mechanical allodynia and downregulates neuroglial activity and the expressions of CGRP and TRPV1 in the spinal segments of a rat model of lumbar disc herniation.

Keyword

GCSB-5; Lumbar disc herniation; Neuropathic pain; Microglia; Astrocytes; Calcitonin gene-related peptide

MeSH Terms

Animals
Anti-Inflammatory Agents, Non-Steroidal
Astrocytes
Calcitonin Gene-Related Peptide
Diagnosis-Related Groups
Ganglia, Spinal
Herbal Medicine*
Horns
Humans
Hyperalgesia
Interleukin-6
Male
Microglia
Models, Animal*
Neuralgia
Polymerase Chain Reaction
Rats*
Rats, Sprague-Dawley
Sciatica
Anti-Inflammatory Agents, Non-Steroidal
Calcitonin Gene-Related Peptide
Interleukin-6

Figure

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  Fig. 1 Study flow schematic.

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  Fig. 2 Changes in mechanical withdrawal thresholds after drug or saline treatment in a rat model of lumbar disc herniation. GCSB-5 or aceclofenac was administered from 3 days after surgery. A significant increase in mean ipsilateral paw withdrawal threshold was found at 56 days after the commencement of GCSB-5 300 treatment versus vehicle controls. *Significantly different from vehicle group, p<0.05.

• 

  Fig. 3 Immunohistochemical expression of Iba1 in the ipsilateral L5 dorsal horn (A) and intensity of Iba1 immunoreaction versus vehicle controls (B) at 56 days after treatment commencement. A : Iba1-positive microglia numbers were lower in the ipsilateral dorsal horns of the GCSB-5 300 group than in the aceclofenac group or vehicle controls. B : the percentage decrease was greater in the GCSB-5 300 group than in the GCSB-5 100, GCSB-5 600, aceclofenac or vehicle control groups. Results are presented as mean±SEM. Bar=100 µm.

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  Fig. 4 Immunohistochemical expression of GFAP in ipsilateral L5 dorsal horns (A) and GFAP immunoreactivities versus vehicle controls (B) at 56 days after treatment commencement. A : GFAP-positive astrocyte numbers were lower in ipsilateral dorsal horns of the GCSB-5 300 group than in the aceclofenac or vehicle controls groups. B : Percentage decreases were greater in the GCSB-5 300 group than in the GCSB-5 100, GCSB-5 600, aceclofenac or vehicle control groups. Results are presented as mean±SEM. Bar=100 µm. GFAP : glial fibrillary acidic protein.

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  Fig. 5 Immunohistochemical expressions of CGRP in ipsilateral L5 dorsal root ganglions (DRGs) (A) and immunoreactivities of CGRP versus the vehicle controls (B) at 56 days after treatment commencement. A : CGRP expression was lower in ipsilateral DRGs in the GCSB-5 300 group than in the aceclofenac or vehicle control groups. B : Percentage decreases were greater in the GCSB-5 300 group than in the GCSB-5 100, GCSB-5 600, aceclofenac or vehicle control groups. Results are presented as mean±SEM. Bar=100 µm. CGRP : calcitonin gene-related peptide.

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  Fig. 6 Immunohistochemical expression of TRPV1 in ipsilateral L5 dorsal root ganglia (DRGs) (A) and TRPV1 immunoreactivities versus the vehicle controls (B) at 56 days after treatment commencement. A : TRPV1 expression was lower in the ipsilateral DRGs of the GCSB-5 300 group than in the aceclofenac or vehicle control groups. B : Percentage decreases in TRPV1 expression were greatest in the GCSB-5 300 group than in the GCSB-5 100, GCSB-5 600, aceclofenac or vehicle control groups. Results are presented as mean±SEM. Bar=100 µm. TRPV1 : transient receptor potential vanilloid 1.

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  Fig. 7 Expressions of IL-6 (A) and CX3CL1 (B) mRNAs in ipsilateral L5 dorsal horns and dorsal root ganglia (DRGs). The mean expressions of the mRNAs of IL-6 and CX3CL1 in the aceclofenac and three GCSB-5 groups were not found to differ significantly from the vehicle control group at 56 days after treatment commencement. Results are presented as mean±SEM. IL-6 : interleukin-6, CX3CL1 : chemokine (C-X3-C motif) ligand 1/fractalkine, mRNA : messenger RNA.

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