Clin Exp Otorhinolaryngol.  2019 Nov;12(4):367-375. 10.21053/ceo.2018.01298.

Response of Glucocorticoid Receptor Alpha and Histone Deacetylase 2 to Glucocorticoid Treatment Predicts the Prognosis of Sudden Sensorineural Hearing Loss

• Zhang X
• Chen J
• Gao Z
• Qi H
• Dai Y
• She W

• KMID: 2462732
• DOI: http://doi.org/10.21053/ceo.2018.01298

Abstract

OBJECTIVES
To investigate glucocorticoid receptor (GR) and histone deacetylase 2 (HDAC2) gene expression and protein levels in peripheral blood mononuclear cells (PBMCs) of patients with severe or profound sudden sensorineural hearing loss (SSNHL) and to explore the roles of GRs and HDAC2 in glucocorticoid (GC) insensitivity.
METHODS
Fifty-five severe or profound SSNHL patients were enrolled in the study. According to hearing improvement after GC treatment, patients were assigned into two groups: GC-sensitive and GC-resistant. A normal reference group included 20 healthy volunteers without hearing loss. Quantitative real-time polymerase chain reaction and Western blot analyses were used to detect the relative expression of GRα, GRβ, and HDAC2 in PBMCs at the mRNA and protein levels.
RESULTS
The protein levels of GRs and HDAC2 in PBMCs of SSNHL patients were lower than the normal reference values before GC treatment. Compared with the GC-resistant group, both the mRNA and protein levels of GRα and HDAC2 were significantly increased in the GC-sensitive group after GC treatment.
CONCLUSION
A lack of GRα and HDAC2 induction following steroid treatment in GC-resistant SSNHL patients may play a fundamental mechanistic role in GC insensitivity. Response of GRα and HDAC2 to steroid treatment may, thus, predict the prognosis of hearing improvement in SSNHL patients.

Keyword

Sudden Sensorineural Loss; Glucocorticoid Receptor; Deacetylase 2; Prognosis

MeSH Terms

Blotting, Western
Gene Expression
Healthy Volunteers
Hearing
Hearing Loss
Hearing Loss, Sensorineural*
Histone Deacetylase 2*
Histone Deacetylases*
Histones*
Humans
Prognosis*
Real-Time Polymerase Chain Reaction
Receptors, Glucocorticoid*
Reference Values
RNA, Messenger
Histone Deacetylase 2
Histone Deacetylases
Histones
RNA, Messenger
Receptors, Glucocorticoid

Figure

• 

  Fig. 1. Glucocorticoid receptor α (GRα) (A) and GRβ mRNA (B) relative expression in the normal reference (NR) group and sudden sensorineural hearing loss patients before treatment. There were no significant differences between groups (all P>0.05). Comparison of GRα (C) and GRβ mRNA (D) relative expression in the glucocorticoid-sensitive (GCS) group and the glucocorticoid-resistant (GCR) group before and after glucocorticoid (GC) treatment. After GC treatment, GRα mRNA expression significantly increased in the GCS group (P<0.001), while no significant change was observed in the GCR group (P=0.91). After GC treatment, slightly increased GRβ mRNA expression was observed in the GCS and the GCR groups, however, there were no significant differences between groups (all P>0.05). (A, C) Comparison between groups using one-way analysis of variance test. (C, D) Comparison before and after treatment using paired-sample test.

• 

  Fig. 2. (A) Histone deacetylase 2 (HDAC2) mRNA relative expression in normal reference (NR) group and sudden sensorineural hearing loss (SSNHL) patients before glucocorticoid (GC) treatment. There were no significant differences between groups. (B) Comparison of HDAC2 mRNA relative expression in glucocorticoid-sensitive (GCS) group and glucocorticoid-resistant (GCR) group before and after GC treatment. After GC treatment, the HDAC2 mRNA expression significantly increased in the GCS group (P<0.001) while no significant change was observed in the GCR group (P=0.52). (A) Comparison between groups using one-way analysis of variance test. (B) Comparison before and after treatment using paired-sample test.

• 

  Fig. 3. Western blotting for glucocorticoid receptor α (GRα) (86 kD) and histone deacetylase 2 (HDAC2; 55 kD) in the normal reference (NR), glucocorticoid-sensitive (GCS) and glucocorticoid-resistant (GCR) groups. Each lane represents one sample from each group. a)Before glucocorticoid treatment. b)After glucocorticoid treatment.

• 

  Fig. 4. (A) Glucocorticoid receptor α (GRα) protein in the normal reference (NR) group and sudden sensorineural hearing loss (SSNHL) patients before glucocorticoid (GC) treatment. Prior to treatment, significantly lower GRα protein levels were observed in the glucocorticoid-sensitive (GCS) and glucocorticoid-resistant (GCR) groups compared to the NR group (P=0.001 or P<0.05), with the lowest GRα protein levels observed in the GCR group. (B) Comparison of GRα protein level in the GCS and GCR groups before and after GC treatment. After GC treatment, GRα protein levels significantly increased in the GCS group (P<0.001), while no significant change was observed in the GCR group (P=0.60). (A) Comparison between groups using one-way analysis of variance test. (B) Comparison before and after treatment using paired-sample test.

• 

  Fig. 5. (A) Histone deacetylase 2 (HDAC2) protein in the normal reference (NR) group and sudden sensorineural hearing loss (SSNHL) patients before glucocorticoid (GC) treatment. Prior to GC treatment, HDAC2 protein levels were significantly lower in SSNHL patients compared to the NR group (P<0.001). (B) Comparison of HDAC2 protein levels in the glucocorticoid-sensitive (GCS) and glucocorticoid-resistant (GCR) groups before and after GC treatment. After glucocorticoid (GC) treatment, HDAC2 protein levels significantly increased in the GCS group (P<0.001), while no significant change was observed in the GCR group (P=0.12). (A) Comparison between groups using one-way analysis of variance test. (B) Comparison before and after treatment using paired-sample test.

• 

  Fig. 6. The comparison of the expression levels of glucocorticoid receptor α (GRα) (A, B) and histone deacetylase 2 (HDAC2) (C, D) between the normal reference (NR) group and the glucocorticoid-sensitive (GCS) group after treatment. After glucocorticoid (GC) treatment, the mRNA level of GRα and HDAC2 in GCS group significantly increased and were higher compared with NR group (P<0.001 or P=0.05). The protein level of GRα in GCS group also significantly increased (P<0.05) and were higher compared with NR group. The protein level of HDAC2 also increased after treatment but were still lower compared with NR group (P<0.05). Comparison between groups using independent samples t-test.

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