Ann Dermatol.  2019 Dec;31(6):611-620. 10.5021/ad.2019.31.6.611.

A Novel Effect of Lipids Extracted from Vernix Caseosa on Regulation of Filaggrin Expression in Human Epidermal Keratinocytes

• Qiao W
• Jia T
• Gu H
• Guo R
• Kaku K
• Wu W

• KMID: 2461983
• DOI: http://doi.org/10.5021/ad.2019.31.6.611

Abstract

BACKGROUND
Vernix caseosa (VC), which is known as a unique human substance, is a biofilm that covers the skin of most human newborns. VC has many biological functions including anti-infective, skin cleansing and skin barrier repair.
OBJECTIVE
In the study, we purpose to investigate the novel effect of lipids extracted from VC on the regulation of filaggrin (FLG) expression and anti-inflammation in normal human epidermal keratinocyte (NHEK) cells.
METHODS
The lipids were extracted by chloroform/methanol (Folch method) and the major properties of fatty acid methyl esters were determined with gas chromatography-mass spectrometer. The relative viability of NHEK cells was evaluated by Cell Counting Kit 8 assay. The related expression of skin barrier protein was accessed with real-time quantitative polymerase chain reaction, Western blot and Immunofluorescence in NHEK cells with or without poly (I:C). Meanwhile, the changes of thymic stromal lymphopoietin (TSLP) and tumor necrosis factor alpha (TNF-α) are analyzed by enzyme-linked immunosorbent assay.
RESULTS
VC lipids mostly contained saturated and branched chains fatty acids. The expression of mRNA and protein of FLG were significantly increased after the supplement with lipid in NHEK cells. Meanwhile, lipids reversed the inhibition of poly (I:C) on FLG. Moreover, lipids suppressed the over secretion of TSLP and TNF-α induced by poly (I:C).
CONCLUSION
These results indicate that lipids extracted from VC has positive effects on the expression of FLG and anti-inflammation, suggesting that lipids of VC may be used for a reference for novel therapeutic method in reducing and remedying skin disease like atopic disease.

Keyword

Fatty acids; Inflammation; Keratinocytes; Vernix caseosa

MeSH Terms

Biofilms
Blotting, Western
Cell Count
Enzyme-Linked Immunosorbent Assay
Esters
Fatty Acids
Fluorescent Antibody Technique
Humans*
Infant, Newborn
Inflammation
Keratinocytes*
Methods
Polymerase Chain Reaction
RNA, Messenger
Skin
Skin Diseases
Tumor Necrosis Factor-alpha
Vernix Caseosa*
Esters
Fatty Acids
RNA, Messenger
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1 The effect of lipids extracted from vernix caseosa (VC) and Poly (I:C) on normal human epidermal keratinocyte (NHEK) cells viability. (A) NHEK cells were exposed on lipids at the different concentrations (400, 200, 100, 50, 25, 12.5, 6.25, 0 µg/ml) for 24 hours. (B) NHEK cells were exposed on poly (I:C) at the different concentrations (400, 200, 100, 50, 25, 12.5, 6.25, 0 µg/ml) for 24 hours. Cell viability was assessed by Cell Counting Kit 8 assay. The results are representative of five independent experiments. Bars means standard deviations. *p<0.05.

• 

  Fig. 2 The positive effect of lipids on filaggrin (FLG) mRNA and protein in normal human epidermal keratinocyte cells. (A) The levels of FLG mRNA increased after treatment with positive control (WY14643) and different lipids under poly (I:C) untreated and poly (I:C) treated condition; real-time quantitative polymerase chain reaction. (B) The FLG protein expression increased after treatment with positive control (WY14643) and lipids; Western blot. (C) Quantitative processing of FLG protein expression. Data are presented in graphical form that shows the fold change compared with control group and relative density. The values are the mean±standard deviation of at least three independent experiments in duplicate. GAPDH: glyceraldehyde 3-phosphate dehydrogenase, NC: normal control. *p<0.05 compared with normal control, **p<0.01, #p<0.05 compared with poly (I:C)-treated group.

• 

  Fig. 3 Lipids improved the expression of filaggrin (FLG) with or without poly (I:C) in normal human epidermal keratinocytes; immunofluorescence ×200. The data are expressed as relative density in graphical form (FLG/glyceraldehyde 3-phosphate dehydrogenase [GADPH]). Bars mean standard deviations. DAPI: 4′,6-diamidino-2-phenylindole. *p<0.05 compared with normal control, **p<0.05 compared with poly (I:C)-treated group, ***p<0.01.

• 

  Fig. 4 (A) mRNA levels of thymic stromal lymphopoietin (TSLP) and tumor necrosis factor alpha (TNF-α) induced by poly (I:C) decreased after treatment with different concentration lipid in normal human epidermal keratinocyte (NHEK) cells; real-time quantitative polymerase chain reaction. (B) The protein levels of TSLP and TNF-α induced by poly (I:C) decreased after treatment with lipid in NHEK cells; enzyme-linked immunosorbent assay. The values are the mean±standard deviation of at least three independent experiments in duplicate. GAPDH: glyceraldehyde 3-phosphate dehydrogenase. *p<0.05, **p<0.01.

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