Ann Dermatol.  2017 Aug;29(4):407-413. 10.5021/ad.2017.29.4.407.

Adiponectin Upregulates Filaggrin Expression via SIRT1-Mediated Signaling in Human Normal Keratinocytes

Affiliations
  • 1Department of Dermatology, Chung-Ang University Hospital, Seoul, Korea. drseo@cau.ac.kr

Abstract

BACKGROUND
Filaggrin (FLG) is the major component of the epidermal granular layer and binds to and condenses the keratin cytoskeleton. FLG thus contributes to cell compaction and serves as a natural moisturizing factor by promoting unfolding and degradation into hygroscopic amino acids. Loss or downregulation of FLG has been shown to result in a weak stratum corneum, which causes water loss and increases the possibility of skin barrier-related seizure. Adiponectin (Acrp30) contributes to the functional recovery of somatic cells, including human normal epidermal keratinocytes (NHEKs).
OBJECTIVE
To investigate the effect of Acrp30 in FLG expression and identifying its signal transduction mechanism.
METHODS
Normal human keratinocytes were treated with Acrp30 and the levels of FLG were examined. Silent mating type information regulation 2 homolog (SIRT)-targeting siRNA and aryl hydrocarbon receptor nuclear translocator (ARNT)-targeting siRNA were used to identify the role of various signal transduction pathway components.
RESULTS
Acrp30 upregulated SIRT1 and ARNT expression in NHEKs, resulting in increased FLG expression. Treatment with both SIRT1-targeting siRNA and ARNT-targeting siRNA blocked Acrp30 stimulation and silenced FLG expression.
CONCLUSION
Adiponectin upregulates FLG expression through a SIRT1-mediated pathway. Our results suggest that Acrp30 is a promising agent for skin barrier permeability improvement.

Keyword

Adiponectin; ARNT; Filaggrin; Keratinocytes; SIRT1

MeSH Terms

Adiponectin*
Amino Acids
Aryl Hydrocarbon Receptor Nuclear Translocator
Cytoskeleton
Down-Regulation
Humans*
Keratinocytes*
Permeability
RNA, Small Interfering
Seizures
Signal Transduction
Skin
Water
Adiponectin
Amino Acids
Aryl Hydrocarbon Receptor Nuclear Translocator
RNA, Small Interfering
Water

Figure

  • Fig. 1 Adiponectin stimulation increases filaggrin (FLG) expression in a time-dependent and dose-dependent manner without affecting the viability. Human normal epidermal keratinocytes (NHEKs) were treated with Acrp30 for 24 h at the indicated concentrations or with 10 µg/ml Acrp30 at the indicated time lines. (A) The MTT assay was performed to determine NHEK viability after Acrp30 treatment. (B, C) Cell lysates were generated with described lysis buffer and silent mating type information regulation 2 homolog (SIRT1), aryl hydrocarbon receptor nuclear translocator (ARNT), and FLG expression level was analyzed by immunoblotting. Representative results are shown from 3 independent experiments. NT: no treatment, O.D: optical density.

  • Fig. 2 Silent mating type information regulation 2 homolog (SIRT1) inhibition blocks Acrp30-induced filaggrin (FLG) upregulation. Human normal epidermal keratinocytes were pretreated with 60 µM sirtinol for 4 h, after which they were treated with 10 µg/ml Acrp30 for 12 h (A) or 24 h (B). Cell lysates were generated with described lysis buffer and analyzed by immunoblotting. The protein amounts were quantified by comparison to β-actin. Immunoblotting was used to analyze the levels of SIRT1, aryl hydrocarbon receptor nuclear translocator (ARNT), and FLG. All blots shown are representative of 3 independent experiments. ***p<0.001 compared to the untreated group, ****p<0.0005 compared to the untreated group. NT: no treatment, ST: sirtinol.

  • Fig. 3 siRNA-mediated silencing of silent mating type information regulation 2 homolog (SIRT1) blocks Acrp30 stimulation and filaggrin (FLG) regulation. Human normal epidermal keratinocytes were transfected with 20 nM of SIRT1-targeting siRNA or scrambled siRNA and incubated for 48 h. Next, the transfected cells were treated with 10 µg/ml Acrp30 for 24 h. Cell lysates were generated with described lysis buffer and analyzed by immunoblotting. The protein amounts were quantified by comparison to β-actin. Immunoblotting was used to analyze the levels of SIRT1, aryl hydrocarbon receptor nuclear translocator (ARNT), and FLG. All blots shown are representative of 3 independent experiments. **p<0.05 compared to the untreated group,. ***p<0.001 compared to the untreated group, ****p <0.0005 compared to the untreated group.

  • Fig. 4 siRNA-mediated knockdown of aryl hydrocarbon receptor nuclear translocator (ARNT) blocks Acrp30 stimulation and filaggrin (FLG) regulation. Human normal epidermal keratinocytes were transfected with 20 nM of ARNT-targeting siRNA or scrambled siRNA and incubated for 48 h. Next, the transfected cells were treated with 10 µg/ml Acrp30 for 24 h. Cell lysates were generated with described lysis buffer and analyzed by immunoblotting. The protein amounts were quantified by comparison to β-actin. Immunoblotting was used to analyze the levels of silent mating type information regulation 2 homolog (SIRT1), ARNT, and FLG. All blots shown are representative results of 3 independent experiments. ***p<0.001 compared to the untreated group.


Cited by  3 articles

The Effect of Adiponectin on the Regulation of Filaggrin Expression in Normal Human Epidermal Keratinocytes
Sun Young Choi, Min Jeong Kim, Ga Ram Ahn, Kui Young Park, Mi-Kyung Lee, Seong Jun Seo
Ann Dermatol. 2018;30(6):645-652.    doi: 10.5021/ad.2018.30.6.645.

Adiponectin Attenuates the Inflammation in Atopic Dermatitis-Like Reconstructed Human Epidermis
Hee-Seok Seo, Ki Hyun Seong, Chang-Deok Kim, Seong Jun Seo, Byung Cheol Park, Myung Hwa Kim, Seung-Phil Hong
Ann Dermatol. 2019;31(2):186-195.    doi: 10.5021/ad.2019.31.2.186.

Adiponectin Promotes Caspase-14 Expression in Normal Human Epidermal Keratinocytes
Sun Young Choi, Min Jeong Kim, Ji Yeon Hong, Kui Young Park, Seong Jun Seo
Ann Dermatol. 2019;31(3):352-355.    doi: 10.5021/ad.2019.31.3.352.


Reference

1. Sandilands A, Sutherland C, Irvine AD, McLean WH. Filaggrin in the frontline: role in skin barrier function and disease. J Cell Sci. 2009; 122:1285–1294.
Article
2. Kezic S, Kammeyer A, Calkoen F, Fluhr JW, Bos JD. Natural moisturizing factor components in the stratum corneum as biomarkers of Filaggrin genotype: evaluation of minimally invasive methods. Br J Dermatol. 2009; 161:1098–1104.
Article
3. Wong WJ, Richardson T, Seykora JT, Cotsarelis G, Simon MC. Hypoxia-inducible factors regulate Filaggrin expression and epidermal barrier function. J Invest Dermatol. 2015; 135:454–461.
Article
4. Ming M, Zhao B, Shea CR, Shah P, Qiang L, White SR, et al. Loss of sirtuin 1 (SIRT1) disrupts skin barrier integrity and sensitizes mice to epicutaneous allergen challenge. J Allergy Clin Immunol. 2015; 135:936–945.e4.
Article
5. Kwon HS, Ott M. The ups and downs of SIRT1. Trends Biochem Sci. 2008; 33:517–525.
Article
6. Guo X, Kesimer M, Tolun G, Zheng X, Xu Q, Lu J, et al. The NAD(+)-dependent protein deacetylase activity of SIRT1 is regulated by its oligomeric status. Sci Rep. 2012; 2:640.
Article
7. Chaudhary N, Pfluger PT. Metabolic benefits from Sirt1 and Sirt1 activators. Curr Opin Clin Nutr Metab Care. 2009; 12:431–437.
Article
8. Blander G, Bhimavarapu A, Mammone T, Maes D, Elliston K, Reich C, et al. SIRT1 promotes differentiation of normal human keratinocytes. J Invest Dermatol. 2009; 129:41–49.
Article
9. Laemmle A, Lechleiter A, Roh V, Schwarz C, Portmann S, Furer C, et al. Inhibition of SIRT1 impairs the accumulation and transcriptional activity of HIF-1α protein under hypoxic conditions. PLoS One. 2012; 7:e33433.
Article
10. Fu Y, Luo N, Klein RL, Garvey WT. Adiponectin promotes adipocyte differentiation, insulin sensitivity, and lipid accumulation. J Lipid Res. 2005; 46:1369–1379.
Article
11. Jin T, Kim OY, Shin MJ, Choi EY, Lee SS, Han YS, et al. Fisetin up-regulates the expression of adiponectin in 3T3-L1 adipocytes via the activation of silent mating type information regulation 2 homologue 1 (SIRT1)-deacetylase and peroxisome proliferator-activated receptors (PPARs). J Agric Food Chem. 2014; 62:10468–10474.
Article
12. Liu Q, Gauthier MS, Sun L, Ruderman N, Lodish H. Activation of AMP-activated protein kinase signaling pathway by adiponectin and insulin in mouse adipocytes: requirement of acyl-CoA synthetases FATP1 and Acsl1 and association with an elevation in AMP/ATP ratio. FASEB J. 2010; 24:4229–4239.
Article
13. Cantó C, Gerhart-Hines Z, Feige JN, Lagouge M, Noriega L, Milne JC, et al. AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity. Nature. 2009; 458:1056–1060.
Article
14. Fullerton MD, Steinberg GR. SIRT1 takes a backseat to AMPK in the regulation of insulin sensitivity by resveratrol. Diabetes. 2010; 59:551–553.
Article
15. Shen Z, Liang X, Rogers CQ, Rideout D, You M. Involvement of adiponectin-SIRT1-AMPK signaling in the protective action of rosiglitazone against alcoholic fatty liver in mice. Am J Physiol Gastrointest Liver Physiol. 2010; 298:G364–G374.
Article
16. Iwabu M, Yamauchi T, Okada-Iwabu M, Sato K, Nakagawa T, Funata M, et al. Adiponectin and AdipoR1 regulate PGC-1alpha and mitochondria by Ca(2+) and AMPK/SIRT1. Nature. 2010; 464:1313–1319.
Article
17. Kovács D, Lovászi M, Póliska S, Oláh A, Bíró T, Veres I, et al. Sebocytes differentially express and secrete adipokines. Exp Dermatol. 2016; 25:194–199.
Article
18. Costa Cdos S, Rohden F, Hammes TO, Margis R, Bortolotto JW, Padoin AV, et al. Resveratrol upregulated SIRT1, FOXO1, and adiponectin and downregulated PPARγ1-3 mRNA expression in human visceral adipocytes. Obes Surg. 2011; 21:356–361.
Article
19. Naowaboot J, Chung CH, Choi R. Rutin stimulates adipocyte differentiation and adiponectin secretion in 3T3-L1 adipocytes. J Med Assoc Thai. 2015; 98:Suppl 3. S1–S6.
20. Furue M, Takahara M, Nakahara T, Uchi H. Role of AhR/ARNT system in skin homeostasis. Arch Dermatol Res. 2014; 306:769–779.
Article
21. van den Bogaard EH, Bergboer JG, Vonk-Bergers M, van Vlijmen-Willems IM, Hato SV, van der Valk PG, et al. Coal tar induces AHR-dependent skin barrier repair in atopic dermatitis. J Clin Invest. 2013; 123:917–927.
Article
22. Denison MS, Nagy SR. Activation of the aryl hydrocarbon receptor by structurally diverse exogenous and endogenous chemicals. Annu Rev Pharmacol Toxicol. 2003; 43:309–334.
Article
23. Wu Z, Uchi H, Morino-Koga S, Shi W, Furue M. Resveratrol inhibition of human keratinocyte proliferation via SIRT1/ARNT/ERK dependent downregulation of aquaporin 3. J Dermatol Sci. 2014; 75:16–23.
Article
24. Schäfer M, Farwanah H, Willrodt AH, Huebner AJ, Sandhoff K, Roop D, et al. Nrf2 links epidermal barrier function with antioxidant defense. EMBO Mol Med. 2012; 4:364–379.
Article
25. Kim EJ, Jeong MS, Li K, Park MK, Lee MK, Yoon Y, et al. Genetic polymorphism of FLG in Korean ichthyosis vulgaris patients. Ann Dermatol. 2011; 23:170–176.
Article
Full Text Links
  • AD
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr