Elsevier

Journal of Voice

Volume 35, Issue 3, May 2021, Pages 342-348
Journal of Voice

Impact of Pregnancy and Glucocorticoid Treatment on NF-κB and MUC5AC in Mucosa of Rat Larynx

https://doi.org/10.1016/j.jvoice.2019.11.008Get rights and content

SUMMARY

Objective

The aim of this study is to reveal physiological expression and distribution of nuclear factor-kappa B (NF-κB) and MUC 5 subtype AC (MUC5AC) in rat laryngeal mucosa and to find out the effect of pregnancy and glucocorticoid treatment on these biomolecules.

Methods

This animal experiment was done in Experimental Animals Research and Application Center of Manisa Celal Bayar University in accordance with the accepted policy on the use of animals. A total of 30 young, adult Wister albino female rats were randomized into a control group (group A), a pregnant group (group B), and a steroid administered group (group C). Sacrification was done by injection of sodium-pentobarbitone (400 mg/kg) solution via intraperitoneal route in all groups. Serum estradiole (E2) and progesterone (PG) were determined by enzyme-linked immunosorbent assay. The relative expression and distribution of NF-κB and MUC5AC in laryngeal mucosa was studied both by immunohistochemistry (IHC) and polymerase chain reaction testing. Expression and immunohistochemical localization of NF-κB and MUC5AC was evaluated by light microscopy (Olympus BX41). In statistical analyses; relative expression of NF-κB and MUC5AC were compared on group basis. The effect of E2 and PG levels on these biomolecules was also evaluated.

Results

NF-κB was found to be significantly low both in group B (P < 0.05) and C (P < 0.001) when compared with group A, while MUC5AC was found to be significantly high both in group B (P < 0.05) and group C (P < 0.05) when compared with group A. Concerning IHC; NF-κB was found to be expressed in epithelium and lamina propria. MUC5AC was found to be expressed particularly in the epithelial layer in all groups. Statistically significant negative correlation between PG and NF-κB expression (P = 0.048), but no correlation between PG and MUC5AC expression (P = 0.487) were revealed. On the other hand, no correlation was found between E2 and the expression of relevant biomolecules (NF-κB [P = 0.270], MUC5AC [P = 0.829]). We also did found a significant negative correlation between the expression of NF-κB and MUC5AC (P = 0.031).

Conclusions

In this study, the physiological expression of NF-κB and MUC5AC in rat laryngeal mucosa was shown for the first time both by polymerase chain reaction and IHC. The impact of pregnancy and glucocorticoid treatment on the expression and distribution of these biomolecules was also revealed. The expression of NF-κB was found to be decreased while the expression of MUC5AC was found to be increased both by pregnancy and glucocorticoid treatment. The inhibitory effect of serum PG on NF-κB expression in rat laryngeal mucosa was also shown for the first time. The expression of MUC5AC was found to be increased both in pregnant and glucocorticoid administered group. Negative correlation between NF-κB and MUC5AC expression was also revealed in rat larynx for the first time. These findings may partially unclose the histochemical background of voice changes caused by pregnancy and as well as by glucocorticoid treatment.

Section snippets

INTRODUCTION

Larynx comes to be known as a secondary sexual organ. Receptors of sex hormones have been shown in laryngeal mucosa.1 Besides, it has been revealed that fluctuation of sex hormone levels due to particular physiologic, pathologic, or iatrogenic conditions may lead explicit changes in quality of voice.2, 3, 4, 5 But, the exact underlying biomolecular and histochemical changes relevant to voice alterations with regard to sex hormones have not been studied thoroughly.

Nuclear factor-kappa B (NF-κB)

Animals

This study was performed in Experimental Animals Research and Application Center of Manisa Celal Bayar University in accordance with the accepted policy on the use of animals. All the procedures were approved by the institutional Animal Ethics Committee.

A total of 30 young, adult (8–12 week old) Wister albino female rats weighing 200–220 g were enrolled into the study. All rats were fed ad libitum and housed at room temperature (22 ± 2°C) on a 12 hours light-dark cycle. All experiments were

PCR and ELISA

A total of 30 Wister albino female rats were included into the experiment (10 control, 10 pregnant, 10 steroid administered). The mean relative expression of mRNA coding for NF-κB of group A, B, and C was; 2.09 ± 0.71, 1.18 ± 0.47, and 0.97 ± 0.28, respectively. The mean relative expression of mRNA coding for MUC5AC of group A, B, and C was; 0.17 ± 0.15, 0.53 ± 0.22, and 1.47 ± 1.03, respectively. The mean serum E2 levels of group A, B and, C was; 21.95 ± 1.35 pg/mL, 73.59 ± 3.18 pg/mL, and

DISCUSSION

It is has been long known that pregnant women suffer from dysphonia to a certain extent.19 Some studies utilizing objective voice analyses support this phenomenon.20, 21, 22, 23 These voice changes can be linked to anatomical alterations (increased level of diaphragm, increased circumference of chest, and widened subcostal angle) and histological changes of laryngeal mucosa which is known to be affected by some hormones.24 During pregnancy, levels of certain hormones (human placental lactogen,

CONCLUSIONS

In this study, the physiological expression of NF-κB and MUC5AC in rat laryngeal mucosa was shown for the first time both by PCR and Immunohistochemistry. The impact of pregnancy and glucocorticoid treatment on the expression and distribution of these biomolecules was also revealed. The expression of NF-κB was found to be decreased while the expression of MUC5AC was found to be increased by pregnancy and as well as by glucocorticoid treatment. Besides, the inhibitory effect of serum PG on NF-κB

CONFLICTS OF INTEREST

The authors declare no conflicts of interest.

REFERENCES (39)

  • E Kalkhoven et al.

    Negative interaction between the RelA(p65) subunit of NF-kappaB and the progesterone receptor

    J Biol Chem

    (1996)
  • YC Zang et al.

    Regulatory effects of estriol on T cell migration and cytokine profile: inhibition of transcription factor NF-kappa B

    J Neuroimmunol

    (2002)
  • G Beauchef et al.

    The p65 subunit of NF-κB inhibits COL1A1 gene transcription in human dermal and scleroderma fibroblasts through its recruitment on promoter by protein interaction with transcriptional activators (c-Krox, Sp1, and Sp3)

    J Biol Chem

    (2012)
  • K Aydin et al.

    Voice characteristics associated with polycystic ovary syndrome

    Laryngoscope

    (2016)
  • C Bultynck et al.

    Self-perception of voice in transgender persons during cross-sex hormone therapy

    Laryngoscope

    (2017)
  • EM Meurer et al.

    Vocal range in the speech of users of low-dose oral contraceptives

    J Voice

    (2017)
  • W Wang et al.

    Mucin 5 subtype AC expression and upregulation in the nasal mucosa of allergic rhinitis rats

    Otolaryngol Head Neck Surg

    (2012)
  • TD Gilmore

    Introduction to NF-kappaB: players, pathways, perspectives

    Oncogene

    (2006)
  • E Dale et al.

    A role for transcription factor NF-kappaB in autoimmunity: possible interactions of genes, sex, and the immune response

    Adv Physiol Educ

    (2006)
  • Cited by (3)

    • Role of VPAC1 and VPAC2 receptors in the etiology of pregnancy rhinitis: an experimental study in rats

      2022, Brazilian Journal of Otorhinolaryngology
      Citation Excerpt :

      The following morning vaginal smears of female rats were examined for sperm existence. We accept the observation of sperm as the day 0 of the gestation as described before.27–29 The first 10 rats with negative and 10 rats with positive vaginal smear were chosen from among 32 rats as the control group (Group A) and the pregnant group (Group B), respectively.

    • The Role of TREK-1 and AQP5 in Gonadocorticoid-Related Voice Disorders

      2022, Journal of Voice
      Citation Excerpt :

      Similar voice alterations have been documented in some other physiological or pathological conditions in which sex hormone levels fluctuate.5,6 But there are very few studies which analyze sex hormone-induced histochemical and biomolecular changes in vocal cord mucosa.1,7-9 TWIK-related potassium channel-1 (TREK-1) is one of the subfamilies of mechano-gated K+ channels.

    View full text