인간성대 섬유아세포주에서 Acid와 Pepsin의 효과
Effects by Acid and Pepsin Exposure in Human Vocal Fold Fibroblast
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Background and Objectives：Laryngopharyngeal reflux, the backflow of gastric contents into the pharynx and larynx, is among the most important factors behind the development of inflammatory disorders of the upper airway. Our objective was to clarify the effects of exposure of the acid and pepsin in the laryngopharyngeal reflux disease by carrying out research on gene changes in the human vocal fold fibroblast. Material and Methods：To make a cell culture solution of acidic conditions of pH4 and 5 by using a 0.01 M hydrochloric acid. Pepsin was prepared in 0.5 mg/mL concentration dissolved in 0.9% sodium chloride. Pepsin was added to the cell culture fluid of pH4 and pH5 acid. The prepared acidic cell culture solution for each condition was treated for 5 minutes on the fibroblasts of the vocal folds. Then to remove the acidic cell culture solution, the vocal fold fibroblast culture fluid was incubated for 6 hours. After the culture, real time PCR, MTT assay and LDH assay were carried out. Results：The human vocal fold fibroblast in the pH4 acidic cell culture medium containing pepsin has the highest cell damage form. We confirmed these results by using the MTT assay and LDH assay. After incubation for 6 hours, it was confirmed that the cell was returned in the normal form of a vocal fold fibroblast in all experimental groups compared to the control groups. There are no significant differences of gene expression after acid and/or pepsin treatment in human vocal fold fibroblast. Conclusions：The most critical effect on the damage to the vocal fold cells was the interaction of pH4 acidic environment and pepsin than acting alone or pH5 environment.
Abstract
Background and Objectives：Laryngopharyngeal reflux, the backflow of gastric contents into the pharynx and larynx, is among the most important factors behind the development of inflammatory disorders of the upper airway. Our objective was to clarify the effects of exposure of the acid and pepsin in the laryngopharyngeal reflux disease by carrying out research on gene changes in the human vocal fold fibroblast. Material and Methods：To make a cell culture solution of acidic conditions of pH4 and 5 by using a 0.01 M hydrochloric acid. Pepsin was prepared in 0.5 mg/mL concentration dissolved in 0.9% sodium chloride. Pepsin was added to the cell culture fluid of pH4 and pH5 acid. The prepared acidic cell culture solution for each condition was treated for 5 minutes on the fibroblasts of the vocal folds. Then to remove the acidic cell culture solution, the vocal fold fibroblast culture fluid was incubated for 6 hours. After the culture, real time PCR, MTT assay and LDH assay were carried out. Results：The human vocal fold fibroblast in the pH4 acidic cell culture medium containing pepsin has the highest cell damage form. We confirmed these results by using the MTT assay and LDH assay. After incubation for 6 hours, it was confirmed that the cell was returned in the normal form of a vocal fold fibroblast in all experimental groups compared to the control groups. There are no significant differences of gene expression after acid and/or pepsin treatment in human vocal fold fibroblast. Conclusions：The most critical effect on the damage to the vocal fold cells was the interaction of pH4 acidic environment and pepsin than acting alone or pH5 environment.
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