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. 2023 Aug 1;13:12491. doi: 10.1038/s41598-023-38828-w

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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(A) Antinociception index obtained from the hot plate assay exploring the pharmacological antagonism of naloxone. The antinociceptive effect of protonectin-F (16 nmol) and morphine (16 nmol) was evaluated after administration of naloxone, as well as protonectin-F or morphine alone. Control groups were treated with vehicle alone or with naloxone and vehicle. Data were analyzed by Two-Way ANOVA followed by Bonferroni’s post-hoc test. (***) indicates statistical difference when compared to vehicle control p < 0.001. (###) indicates difference when compared the group treated with naloxone and morphine 16 nmol with p < 0.001. (+ + +) indicates difference when compared to the control group treated with naloxone and protonectin- F with p < 0.001. (B) Antinociception index obtained from the hot plate assay exploring the pharmacological antagonism of AM251. The antinociceptive effect of protonectin-F was evaluated after administration of AM251, as well as protonectin-F alone. Control groups were treated with vehicle or with AM251. Data were analyzed by Two-Way ANOVA followed by Tukey’s post-hoc test. (***) indicates statistical difference when compared to vehicle control with p < 0.001. (###) indicates difference when compared the group treated with AM251 and protonectin-F with p < 0.001.