OCT4B1 Regulates the Cellular Stress Response of Human Dental Pulp Cells with Inflammation.

Lu Liu,Ruiqi Yang,Xi Wei,Rong Huang

doi:10.1155/2017/2756891

Lu Liu, Ruiqi Yang + Show 2 more

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https://doi.org/10.1155/2017/2756891

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Abstract

Introduction. Infection and apoptosis are combined triggers for inflammation in dental tissues. Octamer-binding transcription factor 4-B1 (OCT4B1), a novel spliced variant of OCT4 family, could respond to the cellular stress and possess antiapoptotic property. However, its specific role in dental pulpitis remains unknown. Methods. To investigate the effect of OCT4B1 on inflammation of dental pulp cells (DPCs), its expression in inflamed dental pulp tissues and DPCs was examined by in situ hybridization, real-time PCR, and FISH assay. OCT4B1 overexpressed DPCs model was established, confirmed by western blot and immunofluorescence staining, and then stimulated with Lipopolysaccharide (LPS). Apoptotic rate was determined by Hoechst/PI staining and FACS. Cell survival rate was calculated by CCK8 assay. Results. In situ hybridization, real-time PCR, and FISH assay revealed that OCT4B1 was extensively expressed in inflamed dental pulp tissues and DPCs with LPS stimulation. Western blot and immunofluorescence staining showed the expression of OCT4B1 and OCT4B increased after OCT4B1 transfection. Hoechst/PI staining and FACS demonstrated that less red/blue fluorescence was detected and apoptotic percentage decreased (3.45%) after transfection. CCK8 demonstrated that the survival rate of pCDH-OCT4B1-flag cells increased. Conclusions. OCT4B1 plays an essential role in inflammation and apoptosis of DPCs. OCT4B might operate synergistically with OCT4B1 to reduce apoptosis.

Highlights

Infection and apoptosis are combined triggers for inflammation in dental tissues
In inflamed dental pulp tissues, odontoblasts were surrounded by locally infiltrated immune cells and disorganized tubular dentin was observed beneath the odontoblasts layer (Figure 1(b))
In situ hybridization assay revealed that, in the noninflamed dental pulp tissues, Octamer-binding transcription factor 4-B1 (OCT4B1) was weakly expressed in normal dental pulp cells (DPCs) (Figures 1(c) and 1(e), ×200 and ×400), whereas OCT4B1 was extensively expressed in both the cytoplasm and nucleus of the inflamed DPCs (Figures 1(d) and 1(f), ×200 and ×400)

Summary

Introduction

Infection and apoptosis are combined triggers for inflammation in dental tissues. Octamer-binding transcription factor 4-B1 (OCT4B1), a novel spliced variant of OCT4 family, could respond to the cellular stress and possess antiapoptotic property. To investigate the effect of OCT4B1 on inflammation of dental pulp cells (DPCs), its expression in inflamed dental pulp tissues and DPCs was examined by in situ hybridization, real-time PCR, and FISH assay. OCT4B1 overexpressed DPCs model was established, confirmed by western blot and immunofluorescence staining, and stimulated with Lipopolysaccharide (LPS). Real-time PCR, and FISH assay revealed that OCT4B1 was extensively expressed in inflamed dental pulp tissues and DPCs with LPS stimulation. Once healing tissues become infected and the inflammatory response persists, an adverse effect on bone regeneration occurs [4,5,6,7]. Exposure to LPS activates the cellular stress response, which results in apoptosis and cell death in dental pulp. It is of great importance to develop the anti-inflammatory strategy during dental regeneration

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Conclusion