Nutr Res Pract.  2022 Feb;16(1):1-13. 10.4162/nrp.2022.16.1.1.

Bitter taste receptors protect against skin aging by inhibiting cellular senescence and enhancing wound healing

• Chung MG
• Kim Y
• Cha YK
• Park TH
• Kim Y

• KMID: 2525510
• DOI: http://doi.org/10.4162/nrp.2022.16.1.1

Abstract

BACKGROUND/OBJECTIVES
Bitter taste receptors are taste signaling pathway mediators, and are also expressed and function in extra-gustatory organs. Skin aging affects the quality of life and may lead to medical issues. The purpose of this study was to better understand the anti-skin aging effects of bitter taste receptors in D-galactose (D-gal)-induced aged human keratinocytes, HaCaT cells.
MATERIALS/METHODS
Expressions of bitter taste receptors in HaCaT cells and mouse skin tissues were examined by polymerase chain reaction assay. Bitter taste receptor was overexpressed in HaCaT cells, and D-gal was treated to induce aging. We examined the effects of bitter taste receptors on aging by using β-galactosidase assay, wound healing assay, and Western blot assay.
RESULTS
TAS2R16 and TAS2R10 were expressed in HaCaT cells and were upregulated by D-gal treatment. TAS2R16 exerted protective effects against skin aging by regulating p53 and p21, antioxidant enzymes, the SIRT1/mechanistic target of rapamycin pathway, cell migration, and epithelial-mesenchymal transition markers. TAS2R10 was further examined to confirm a role of TAS2R16 in cellular senescence and wound healing in D-gal-induced aged HaCaT cells.
CONCLUSIONS
Our results suggest a novel potential preventive role of these receptors on skin aging by regulating cellular senescence and wound healing in human keratinocyte, HaCaT.

Keyword

Taste; galactose; skin aging; wound healing; keratinocytes

Figure

• 

  Fig. 1 Expression of bitter taste receptors in HaCaT cells and mouse skin tissues. (A) The mRNA levels of human bitter taste receptors TAS2R16 and TAS2R10 in D-gal-induced aged HaCaT cells. GAPDH was used as a loading Ctrl. (B) The mRNA levels of mouse bitter taste receptors Tas2r118 and Tas2r114 in D-gal-induced aged mouse model. Gapdh was used as a loading Ctrl. (C) Protein expression of TRPM5 and PLCβ2 in D-gal-induced aged HaCaT cells. Quantified results of TRPM5 and PLCβ2 are shown. β-actin was used as a loading Ctrl. A P-value lower than 0.05 was considered statistically significant.Ctrl, control; D-gal, D-galactose; mRNA, messenger RNA.*P < 0.05, **P < 0.01

• 

  Fig. 2 Effects of TAS2R16 on aging in D-gal-induced aged HaCaT cells. (A) Transfection efficiency of TAS2R16 in HaCaT cells. β-actin was used as a loading control. (B) SA-β-gal assays to evaluate for cellular senescence. Representative photographs from each EV Ctrl, EV Ctrl + D-gal 200 mM and TAS2R16 + D-gal 200 mM group are shown in the left panel. The quantified result of SA-β-gal assay is shown in the right panel. (C) The protein levels of p53 and p21. (a) Representative bands (left panel) and quantified results of p53 and p21 (right panel) are shown. (D) The protein levels of antioxidant enzymes. Representative bands (left panel) and quantified results of SOD1, GPX1, and catalase (right panel) are shown. (E) Effects of TAS2R16 on the SIRT/mTOR pathway. Representative bands (left panel) and quantified results of SIRT1 and mTOR phosphorylation levels (right panel) are shown. β-actin was used as a loading control. The letters (a, b, and c) indicate statistical significance among the different groups.EV Ctrl, empty vector control; D-gal, D-galactose; SOD1, superoxide dismutase 1; GPX1, glutathione peroxidase 1; mTOR, mechanistic target of rapamycin; p-mTOR, phospho-mechanistic target of rapamycin; SA-β-gal, senescence-associated β-galactosidase.

• 

  Fig. 3 Effects of TAS2R16 on wound healing in D-galactose-induced aged HaCaT cells. (A) Wound healing assays. Representative pictures are shown in the left panel. Quantified results of relative wound width at 24 h and 48 h are shown in the right panel. (B) Wound healing-related markers were examined by Western blot. Representative images (left panel) and quantified results of MMP-2, MMP-9, TIMP-1, TIMP-2, E-cadherin, N-cadherin, and vimentin (right panel). β-actin was used as a loading control. The letters (a, b, and c) stand for statistical significance among the different groups.EV Ctrl, empty vector control; MMP, matrix metalloproteinase; TIMP, tissue inhibitor of metalloproteinase.

• 

  Fig. 4 Effects of TAS2R10 on D-gal-induced aged HaCaT cells. (A) Transfection efficiency of TAS2R10 in HaCaT cells. β-actin was used as a loading control. (B) SA-β-gal assays to evaluate for cellular senescence. Representative pictures from each EV Ctrl, EV Ctrl + D-gal 200 mM and TAS2R10 + D-gal 200 mM group are shown in the left panel. The quantified result of SA-β-gal assay is shown in the right panel. (C) Wound healing assays. Representative pictures are shown in the left panel. Quantified results of relative wound width at 24 h and 48 h are shown in the right panel. Letters (a, b, and c) stand for statistical significance among the different groups.EV Ctrl, empty vector control; SA-β-gal, senescence-associated β-galactosidase.

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