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. 2022 Nov 29;27(23):8321.
doi: 10.3390/molecules27238321.

Effects of Different Opioid Drugs on Oxidative Status and Proteasome Activity in SH-SY5Y Cells

Laura Rullo  1 , Francesca Felicia Caputi  1 , Loredana Maria Losapio  1 , Camilla Morosini  1 , Luca Posa  2 , Donatella Canistro  1 , Fabio Vivarelli  1 , Patrizia Romualdi  1 , Sanzio Candeletti  1
Affiliations

Effects of Different Opioid Drugs on Oxidative Status and Proteasome Activity in SH-SY5Y Cells

Laura Rullo et al. Molecules. .

Abstract

Opioids are the most effective drugs used for the management of moderate to severe pain; however, their chronic use is often associated with numerous adverse effects. Some results indicate the involvement of oxidative stress as well as of proteasome function in the development of some opioid-related side effects including analgesic tolerance, opioid-induced hyperalgesia (OIH) and dependence. Based on the evidence, this study investigated the impact of morphine, buprenorphine or tapentadol on intracellular reactive oxygen species levels (ROS), superoxide dismutase activity/gene expression, as well as β2 and β5 subunit proteasome activity/biosynthesis in SH-SY5Y cells. Results showed that tested opioids differently altered ROS production and SOD activity/biosynthesis. Indeed, the increase in ROS production and the reduction in SOD function elicited by morphine were not shared by the other opioids. Moreover, tested drugs produced distinct changes in β2(trypsin-like) and β5(chymotrypsin-like) proteasome activity and biosynthesis. In fact, while prolonged morphine exposure significantly increased the proteolytic activity of both subunits and β5 mRNA levels, buprenorphine and tapentadol either reduced or did not alter these parameters. These results, showing different actions of the selected opioid drugs on the investigated parameters, suggest that a low µ receptor intrinsic efficacy could be related to a smaller oxidative stress and proteasome activation and could be useful to shed more light on the role of the investigated cellular processes in the occurrence of these opioid drug side effects.

Keywords: ROS; SOD; buprenorphine; morphine; proteasome; tapentadol.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Figure 1
Figure 1
ROS production after treatment with 10 µM morphine (M), 0.25 µM buprenorphine (B) or 10 µM tapentadol (T) in SH-SY5Y cells at different time points (a–d). The right framed insert reports ROS production after cotreatment with 10 µM naloxone (Nal) + 10 µM morphine (M) or 10 µM β-funaltrexamine (β-FNA) + 10 µM morphine (M) in SH-SY5Y cells at 24 h. Data, reported as the mean ± SEM of four/six biological replicates for each treatment, are expressed as the percentage of relative fluorescence (a–d) or the percentage of the intensity of the first spectra line (insert) compared to control (C) and are analyzed by one-way ANOVA followed by Dunnett’s multiple comparisons test (* p < 0.05; ** p < 0.01 vs. control).
Figure 2
Figure 2
SOD activity after treatment with 10 µM morphine (M), 0.25 µM buprenorphine (B) or 10 µM tapentadol (T) in SH-SY5Y cells at 24 h (a) and 48 h (b). Data, reported as the mean ± SEM of three/four biological replicates for each treatment, are expressed as the percentage of inhibition rate (absorbance) compared to control (C) and analyzed by one-way ANOVA followed by Dunnett’s multiple comparisons test (* p < 0.05 vs. control).
Figure 3
Figure 3
SOD1 relative gene expression after treatment with 10 µM morphine (M), 0.25 µM buprenorphine (B) or 10 µM tapentadol (T) in SH-SY5Y cells at 24 h (a) and 48 h (b). Data, expressed as the mean ± SEM of three/four biological replicates for each treatment, represent 2−DDCt values and are analyzed by one-way ANOVA followed by Dunnett’s multiple comparisons test (**** p < 0.0001 vs. control (C)).
Figure 4
Figure 4
β2 trypsin-like (a,b) and β5 chemotrypsin-like (c,d) activities after treatment with 10 µM morphine (M), 0.25 µM buprenorphine (B) or 10 µM tapentadol (T) in SH-SY5Y cells at 24 h (a,c) and 48 h (b,d). Data, reported as the mean ± SEM of three/four biological replicates for each treatment, are expressed as the percentage of relative fluorescence (RFU) compared to control (C) and analyzed by one-way ANOVA followed by Dunnett’s multiple comparisons test (** p < 0.01; *** p < 0.001 vs. control; **** p < 0.0001 vs. control).
Figure 5
Figure 5
Relative gene expression of β2 (a,b) and β5 (c,d) subunits after treatment with 10 µM morphine (M), 0.25 µM buprenorphine (B) or 10 µM tapentadol (T) in SH-SY5Y cells at 24 h (a,c) and 48 h (b,d). Data, expressed as the mean ± SEM of three/four biological replicates per treatment, represent 2−DDCt values and are analyzed by one-way ANOVA followed by Dunnett’s multiple comparisons test (* p < 0.05; ** p < 0.01; *** p < 0.001 vs. control (C)).
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