Abstract
Spinal cord injury (SCI) is a deliberating disorder with impairments in locomotor deficits and incapacitating sensory abnormalities. Harpagophytum procumbens (Hp) is a botanical widely used for treating inflammation and pain related to various inflammatory and musculoskeletal conditions. Using a modified rodent contusion model of SCI, we explored the effects of this botanical on locomotor function and responses to mechanical stimuli, and examined possible neurochemical changes associated with SCI-induced allodynia. Following spinal cord contusion at T10 level, Hp (300 mg/kg, p.o.) or vehicle (water) was administered daily starting 24 h post-surgery, and behavioral measurements made every-other day until sacrifice (Day 21). Hp treatment markedly ameliorated the contusion-induced decrease in locomotor function and increased sensitivity to mechanical stimuli. Determination of Iba1 expression in spinal cord tissues indicated microglial infiltration starting 3 days post-injury. SCI results in increased levels of 4-hydroxynonenal, an oxidative stress product and proalgesic, which was diminished at 7 days by treatment with Hp. SCI also enhanced antioxidant heme oxygenase-1 (HO-1) expression. Concurrent studies of cultured murine BV-2 microglial cells revealed that Hp suppressed oxidative/nitrosative stress and inflammatory responses, including production of nitric oxide and reactive oxygen species, phosphorylation of cytosolic phospholipases A2, and upregulation of the antioxidative stress pathway involving the nuclear factor erythroid 2-related factor 2 and HO-1. These results support the use of Hp for management of allodynia by providing resilience against the neuroinflammation and pain associated with SCI and other neuropathological conditions.
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Abbreviations
- 4-HHE:
-
4-Hydroxyhexenal
- 4-HNE:
-
4-Hydroxynonenal
- ARA:
-
Arachidonic acid
- CHD:
-
1,3-Cyclohexanedione
- COX-1/2:
-
Cyclooxygenase-1/2
- cPLA2 :
-
Cytosolic phospholipases A2
- DHA:
-
Docosahexaenoic acid
- DCF:
-
ROS detection reagent CM-H2DCFDA
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- ESI:
-
Electrospray ionization
- FBS:
-
Fetal bovine albumin
- HO-1:
-
Heme oxygenase-1
- Hp:
-
Harpagophytum procumbens, or Devil’s Claw
- HPLC:
-
Ammonium acetate
- Ibu:
-
Ibuprofen
- iNOS:
-
Nitric oxide synthase
- LPS:
-
Lipopolysaccharide
- Nrf2:
-
Erythroid 2-related factor 2
- NQO1:
-
NAD(P)H quinone dehydrogenase 1
- ROS:
-
Reactive oxygen species
- SCI:
-
Spinal cord injury
- SPE:
-
Solid phase extraction
- TBS-T:
-
Tween 20
- TOF–MS/MS:
-
Time-of-flight tandem mass spectrometry
- UHPLC:
-
Ultra-high performance liquid chromatography
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Acknowledgements
We thank Dr. Brian Mooney, associate director of the Charles W. Gehrke Proteomics Center at the University of Missouri, for providing assistance with the LC–MS/MS, and Dr. Lloyd W. Sumner, Professor of Biochemistry and Director of MU Metabolomics Center for UHPLC analysis of H. procumbens extract. Thanks are due to Professor Xiaoming Xu at Department of Neurosurgery, Stark Neurosciences Research Institute, Indiana University for providing advise for Western blot analysis of p-cPLA2 and cPLA2 in spinal cord tissue.
Funding
This work was supported in part by the funding from the Missouri Spinal Cord Injury/Disease Research Program (SCIDRP), MIZZOU Advantage grant and research funds of the University of Missouri (ZG), and in part, support by the NIH/NCCIH (R21 AT009086). The contents are solely the responsibility of the authors and do not necessarily represent the official views of the sponsors.
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Ungerer, G., Cui, J., Ndam, T. et al. Harpagophytum procumbens Extract Ameliorates Allodynia and Modulates Oxidative and Antioxidant Stress Pathways in a Rat Model of Spinal Cord Injury. Neuromol Med 22, 278–292 (2020). https://doi.org/10.1007/s12017-019-08585-z
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DOI: https://doi.org/10.1007/s12017-019-08585-z