Korean J Pain.  2024 Jul;37(3):233-246. 10.3344/kjp.24042.

Ferroptosis inhibitor ferrostatin-1 attenuates morphine tolerance development in male rats by inhibiting dorsal root ganglion neuronal ferroptosis

• Dirik H
• Taşşkıran A
• Joha Z

• KMID: 2557727
• DOI: http://doi.org/10.3344/kjp.24042

Abstract

Background
Ferrostatin-1 and liproxstatin-1, both ferroptosis inhibitors, protect cells. Liproxstatin-1 decreases morphine tolerance. Yet, ferrostatin-1's effect on morphine tolerance remains unexplored. This study aimed to evaluate the influence of ferrostatin-1 on the advancement of morphine tolerance and understand the underlying mechanisms in male rats.
Methods
This experiment involved 36 adult male Wistar albino rats with an average weight ranging from 220 to 260 g. These rats were categorized into six groups: Control, single dose ferrostatin-1, single dose morphine, single dose ferrostatin-1 + morphine, morphine tolerance (twice daily for five days), and ferrostatin-1 + morphine tolerance (twice daily for five days). The antinociceptive action was evaluated using both the hot plate and tail-flick tests. After completing the analgesic tests, tissue samples were gathered from the dorsal root ganglia (DRG) for subsequent analysis. The levels of glutathione, glutathione peroxidase 4 (GPX4), and nuclear factor erythroid 2-related factor 2 (Nrf2), along with the measurements of total oxidant status (TOS) and total antioxidant status (TAS), were assessed in the tissues of the DRG.
Results
After tolerance development, the administration of ferrostatin-1 resulted in a significant decrease in morphine tolerance (P < 0.001). Additionally, ferrostatin-1 treatment led to elevated levels of glutathione, GPX4, Nrf2, and TOS (P < 0.001), while simultaneously causing a decrease in TAS levels (P < 0.001).
Conclusions
The study found that ferrostatin-1 can reduce morphine tolerance by suppressing ferroptosis and reducing oxidative stress in DRG neurons, suggesting it as a potential therapy for preventing morphine tolerance.

Keyword

Ferroptosis; Ganglia, Spinal; Glutathione; Morphine; Oxidative Stress; Pain; Phospholipid Hydroperoxide Glutathione Peroxidase

Figure

• 

  Fig. 1 Experimental design of the study. DMSO: dimethyl sulfoxide (ferrostatin dissolved in dimethyl sulfoxide but morphine dissolved in saline), i.p.: intraperitoneally, s.c.: subcutaneousl, GPX4: glutathione peroxidase 4, Nrf2: nuclear factor erythroid 2-related factor 2, TAS: total antioxidant status, TOS: total oxidant status.

• 

  Fig. 2 This figure illustrates the impact of ferrostatin-1 on nociception and morphine analgesia in the tail flick (A) and hot plate (B) tests. The values are presented as the means ± standard error of the mean of % MPE (n = 6). **P < 0.01, indicating a significant difference compared to the control group.

• 

  Fig. 3 This figure depicts the impact of ferrostatin-1 on the development of morphine tolerance in the tail-flick (A) and hot plate (B) tests. The values are presented as the means ± standard error of the mean of % MPE (n = 6). **P < 0.01 indicate a significant difference compared to the control group. ++P < 0.01 indicate a significant difference compared to the morphine group. ##P < 0.01 indicate a significant difference compared to the morphine tolerance group.

• 

  Fig. 4 This figure illustrates the impact of ferrostatin-1 on the levels of glutathione (A), GPX4 (B), and Nrf2 (C) in the dorsal root ganglion during morphine analgesia and tolerance. Values are presented as the means ± standard error of the mean of six rats. GPX4: glutathione peroxidase 4, Nrf2: nuclear factor erythroid 2-related factor 2. ***P < 0.001, compared to the control group. +P < 0.001, compared to the morphine group. #P < 0.001, compared to the morphine tolerance group.

• 

  Fig. 5 This figure illustrates the impact of ferrostatin-1 on TAS (A) and TOS (B) in morphine analgesia and tolerance in dorsal root ganglion. Values are presented as the means ± standard error of the mean of six samples. TAS: total antioxidant status, TOS: total oxidant status. *P < 0.05, **P < 0.01 and ***P < 0.001, compared to the control group. +P < 0.01 and ++P < 0.001, compared to the morphine group. #P < 0.001, compared to the morphine tolerance group.

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