Korean J Physiol Pharmacol.  2024 Jan;28(1):73-81. 10.4196/kjpp.2024.28.1.73.

Naringenin modulates GABA mediated response in a sexdependent manner in substantia gelatinosa neurons of trigeminal subnucleus caudalis in immature mice

• Park SA
• Nguyen TTP
• Park SJ
• Han SK

• KMID: 2550290
• DOI: http://doi.org/10.4196/kjpp.2024.28.1.73

Abstract

The substantia gelatinosa (SG) within the trigeminal subnucleus caudalis (Vc) is recognized as a pivotal site of integrating and modulating afferent fibers carrying orofacial nociceptive information. Although naringenin (4',5,7-thrihydroxyflavanone), a natural bioflavonoid, has been proven to possess various biological effects in the central nervous system (CNS), the activity of naringenin at the orofacial nociceptive site has not been reported yet. In this study, we explored the influence of naringenin on GABA response in SG neurons of Vc using whole-cell patch-clamp technique. The application of GABA in a bath induced two forms of GABA responses: slow and fast. Naringenin enhanced both amplitude and area under curve (AUC) of GABA-mediated responses in 57% (12/21) of tested neurons while decreasing both parameters in 33% (7/21) of neurons. The enhancing or suppressing effect of naringenin on GABA response have been observed, with enhancement occurring when the GABA response was slow, and suppression when it was fast. Furthermore, both the enhancement of slower GABA responses and the suppression of faster GABA responses by naringenin were concentration dependent. Interestingly, the nature of GABA response was also found to be sex-dependent. A majority of SG neurons from juvenile female mice exhibited slower GABA responses, whereas those from juvenile males predominantly displayed faster GABA responses. Taken together, this study indicates that naringenin plays a partial role in modulating orofacial nociception and may hold promise as a therapeutic target for treating orofacial pain, with effects that vary according to sex.

Keyword

GABA receptor; Orofacial nociception; Sex; Substantia gelatinosa neurons; 4’,5,7-thrihydroxyflavanone (naringenin)

Figure

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  Fig. 1 Structures of the flavone and naringenin.

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  Fig. 2 Effects of naringenin and GABA-mediated responses of substantia gelatinosa (SG) neurons under a voltage clamp mode. (A) A representative trace showing the effect of naringenin in a dose-dependent manner (1 μM, 10 μM, 100 μM, 1 mM, and 2 mM, n = 10). (B) A histogram illustrates a weak inward current observed above 1 mM concentration of naringenin. (C, D) A representative trace and histogram show no discernible effect of 0.5 mM naringenin on membrane current (n = 3). (E–H) Representative traces and histograms showing no desensitization of slow (E, G, n = 15) and fast (F, H, n = 6) GABA responses upon repeated application to SG neurons. (I) A histogram demonstrates no significant differences in the response mediated by GABA upon repeated application (total n = 21, paired t-test, p > 0.05, ns: not significant).

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  Fig. 3 Effects of naringenin on GABA-mediated responses of substantia gelatinosa (SG) neurons under a voltage clamp mode. (A, D) Representative current traces illustrate the effect of naringenin on 30 μM GABA-induced inward slow and fast responses, respectively. (B, C) Histograms display a significant enhancement in the mean relative amplitude and area under curve (AUC) of GABA-induced slow responses by 0.5 mM naringenin (n = 12), respectively. (E, F) Histograms showing significant suppression in the mean relative amplitude and AUC of GABA-induced fast responses by 0.5 mM naringenin (n = 7), respectively (paired t-test, *p < 0.05, **p < 0.01, ***p < 0.001).

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  Fig. 4 Dose-dependent effect of naringenin on GABA-mediated slow and fast responses of substantia gelatinosa (SG) neurons. (A, B) Representative traces showing dose-dependent effect of naringenin (1 μM, 10 μM, 100 μM, and 1 mM) on 30 μM GABA-induced slow and fast inward current, respectively. (C) A histogram compares the increasing tendency in the mean relative amplitude of 30 μM GABA-mediated slow responses in the presence of various concentrations of naringenin (n = 4). (D) A histogram compares the decreasing tendency in the mean relative amplitude of 30 μM GABA-mediated fast responses in the presence of various concentrations of naringenin (n = 5) (*p < 0.05, **p < 0.01, one-way ANOVA post-hoc Scheffe test).

• 

  Fig. 5 Effects of naringenin on GABA-mediated responses of substantia gelatinosa (SG) neurons under a current clamp mode. (A) Representative current traces demonstrate the effects of naringenin on 30 μM GABA-induced depolarized responses. (B) A histogram indicates no significant difference in GABA-induced depolarization in the presence of 0.5 mM naringenin (n = 5). (C) A histogram displays a significant increase in the relative area under the curve (AUC) of GABA-induced depolarization in the presence of 0.5 mM naringenin (n = 5) (Paired t-test, nsp > 0.05, *p < 0.05, ns: not significant, respectively). RMP, resting membrane potential; Cc, current clamp.

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  Fig. 6 Histogram depicting the varied nature of GABA responses in male and female juvenile substantia gelatinosa (SG) neurons. (A) Histogram illustrating the percentage of varied GABA response nature across male and female juvenile SG neurons. (B, C) Histogram indicating the percentage of varied effects of naringenin on GABA-mediated inward current and area under the curve (AUC) across male and female, rerspectively (chi-square test, *p < 0.05).

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