Background & Aims

Neuropathic pain, a form of chronic pain that arises from damage or disease of the somatosensory system, is particularly challenging to manage. Despite the variability of pharmacological approaches none of them offer complete relief, and some induce undesirable effects. Consequently, it is necessary a constant chase for novel pharmacological and non-pharmacological approaches to address this issue. An alternative approach is the use of integrative and complementary practices like pharmacoacupuncture – the application of compounds at acupoints. This technique combined with apitoxin injection at the E36 acupoint is attracting attention, however limited research examines its effectiveness and analgesic mechanisms. Our goal was to assess apitoxin’s analgesic effect on neuropathic pain, to evaluate if melittin, apitoxin’s major component, can replicate it, to explore involved pathways, and to study muscle tissue changes post-toxin injections.

Methods

Neuropathy was induced in rats by the chronic constriction of the sciatic nerve (CEUA 7835170522). On the 15th day after surgery, pharmacoacupuncture was performed with either apitoxin or melittin in a single application Mechanical and cold allodynia were evaluated before and at several periods after the pharmacoacupuncture until the analgesic effect is lost. Naloxone, a non-selective opioid receptor antagonist, was used to investigate the involvement of endogenous opioid pathways. Tissue samples were collected on the 19th or 20th day after pharmacoacupuncture to analyze the expression of IBA-1 and GFAP, markers of microglia and astrocytes, and ATF3, which play a crucial role in neuroinflammation and peripheral lesions. Also, on day 20th, acupoint tissue was collected to realize histological analysis and investigate the myofiber cross-sectional area (CSA) after treatments.

Results

Pharmacoacupuncture with apitoxin or melittin significantly increased the mechanical and thermal nociceptive threshold of the animals, effect which remains for 96 or 120 hours after treatment, respectively. Western blotting analysis demonstrates that pharmacoacupuncture with apitoxin or melittin led to a reduction in the expression of ATF3, GFAP, and IBA-1 proteins in both times of investigation. Pretreatment with naloxone blocked the analgesic effect, indicating the involvement of endogenous opioid peptides. Furthermore, in histological assessment, there was an increase of CSA in the groups that received toxin when compared to the saline group. Besides that, we also detected an increase in mast cell degranulation, which may be associated with the therapeutic action of pharmacoacupuncture.

Conclusions

Pharmacoacupuncture with both compounds, apitoxin or melittin, appears as an attractive therapy for chronic pain control due to its efficacy and prolonged duration of pain relief. Also, pharmacoacupuncture can modulate important markers of inflammation in the peripheral and central nervous system. It was also possible to determine that the analgesia promoted by pharmacoacupuncture is mediated via the endogenous opioid pathway and that the treatment can recover the CSA, increasing the capacity for post-injury recovery.

Financial support: Fapesp 2013/07467-1, CAPES and Butantan Foundation.

References

Bennett, GJ; Xie, Y.-K. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 33(1):p 87-107, April 1988. | DOI: 10.1016/0304-3959(88)90209-6.

Kang SY, Roh DH, Yoon SY, Moon JY, Kim HW, Lee HJ, Beitz AJ, Lee JH. Repetitive treatment with diluted bee venom reduces neuropathic pain via potentiation of locus coeruleus noradrenergic neuronal activity and modulation of spinal NR1 phosphorylation in rats. J Pain. 2012 Feb;13(2):155-66. doi: 10.1016/j.jpain.2011.10.012. Epub 2012 Jan 3. PMID: 22217441.

Presenting Author

Ana Maria Boaventura de Oliveira

Poster Authors

Ana Boaventura de Oliveira

Bsc

Butantan Institute

Lead Author

Douglas Felipe Silva

Bsc

Laboratory of Pain and Signaling, Butantan Institute

Lead Author

Morena Brazil Sant’anna

PhD

Laboratory of Pain and Signaling, Butantan Institute

Lead Author

Tamires Cunha Almeida

PhD

Laboratory of Pain and Signaling, Butantan Institute

Lead Author

Gessica Sabrina de Assis Silva

Master

Laboratory of Pain and Signaling, Butantan Institute

Lead Author

Rafael Marques-Porto

PhD

Laboratory of Development and Innovation, Butantan Institute

Lead Author

Pedro Luiz Mailho -Fontana

PhD

Laboratory of Structural Biology, Butantan Institute

Lead Author

Josie Resende da Silva

PhD

Laboratory of Neuroscience, Neuromodulation and Study of Pain - Federal University of Alfenas

Lead Author

Gisele Picolo

Butantan Institute

Lead Author

Topics

  • Models: Chronic Pain - Neuropathic