Screening and Research on Skin Barrier Damage Protective Efficacy of Different Mannosylerythritol Lipids
Abstract
:1. Introduction
2. Results
2.1. Isolation and TLC Verification of MELs
2.2. Quantitative Detection of MELs
2.3. Physicochemical Characterisation of MELs
2.4. Effect of MEL and UVB on Cell Viability in HaCaT Cells
2.5. Effect of MELs on Contents of FLG and TGM1 in UVB-Induced HaCaT Cells
2.6. Increased mRNA Expression of Genes Associated with Skin Barrier in UVB-Induced HaCaT Cells after MEL-B Treatment
2.7. Upregulation of MEL-B Treatment on Expression Levels of LOR, FLG, TGM1 Protein in UVB-Induced HaCaT Cells
2.8. Protective Effects of MEL-B on Skin Barrier Damage as Assessed in SDS-Induced EpiKutis®3D Human Skin Cells
2.9. Effects of MEL-B Treatment on Expression Levels of LOR, FLG, and TGM1 in Cells of the SDS-Induced EpiKutis®3D Human Skin Cell Damage Model
3. Discussion
4. Materials and Methods
4.1. Chemicals and Reagents
4.2. General Experimental Procedures
4.2.1. Production of MELs
4.2.2. Preliminary Separation and Purification of Fermentation Broth
4.2.3. Separation and Purification of MELs by Silica Gel Column Chromatography
4.2.4. Quantification of MELs by High-Performance Liquid Chromatography (HPLC)
4.2.5. Structure Analysis of MELs by HRESIMS and NMR
4.3. Bioactivity Assay
4.3.1. Cell Culture
4.3.2. UVB Irradiation and Treatments
4.3.3. Cell Viability Assay
4.3.4. ELISA
4.3.5. Quantitative Real-Time PCR (RT-PCR) Analysis
4.3.6. Western Blotting
4.3.7. Immunohistochemistry
4.3.8. Immunofluorescence
4.3.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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MEL-A a | MEL-B a | MEL-C b | ||||
---|---|---|---|---|---|---|
Functional Groups | 1H NMR δ (ppm) | 13C NMR δ (ppm) | 1H NMR δ (ppm) | 13C NMR δ (ppm) | 1H NMR δ (ppm) | 13C NMR δ (ppm) |
D-Mannose | ||||||
H-1′ (C-1′) | 5.50, d | 99.35 | 5.50, dd | 99.39 | 5.50, dd | 99.14 |
H-2′ (C-2′) | 5.06, dd | 68.56 | 4.91, dd | 68.74 | 5.10, dd | 68.70 |
H-3′ (C-3′) | 5.24, t | 70.60 | 4.77, m | 71.23 | 5.18, t | 70.64 |
H-4′ (C-4′) | 4.72, d | 65.90 | 3.98, m | 65.64 | 4.77, d | 66.19 |
H-5′ (C-5′) | 3.69, m | 72.55 | 3.85, m | 74.58 | 4.04, m | 74.88 |
H-6′ (C-6′) a | 4.19, dd | 62.40 | 3.81, m | 63.15 | 3.77, m | 61.42 |
H-6′ (C-6′) b | 4.24, dd | 62.40 | 4.43, m | 63.15 | 3.79, m | 61.42 |
meso-Erythritol | ||||||
H-1 (C-1) | 3.76, m | 63.60 | 3.72, m | 63.63 | 3.73, m | 63.48 |
H-2 (C-2) | 3.66, m | 71.86 | 3.66, m | 73.12 | 3.65, m | 73.10 |
H-3 (C-3) | 3.72, m | 71.19 | 3.77, m | 71.93 | 3.75, m | 71.10 |
H-4a (C-4a) | 3.84, dd | 72.38 | 3.56, m | 72.23 | 3.68, m | 71.99 |
H-4b (C-4b) | 3.99, dd | 72.38 | 3.75, m | 72.23 | 3.54, m | 71.99 |
Acetyl group(s) | ||||||
-CH3 | 2.09, 2.03, s | (20.75, 20.68) | 2.13, s | 20.86 | 2.06, s | 20.73 |
-C=O | (170.76, 169.46) | 171.67 | 170.21 | |||
Acyl groups | ||||||
-CH3 | 0.86, m | (14.12, 14.11) | 0.87, m | (14.12, 14.07) | 0.88, m | (14.13, 14.07) |
-(CH2)n- | 1.23–1.31, brs | (22.67–31.88) | 1.24–1.26, brs | (22.61–31.94) | 1.25–1.29, brs | (22.68–31.90) |
-CO-CH2- (C-2′) position | 2.42, m | 34.16 | 2.40, m | 34.11 | 2.43, m | 34.11 |
-CO-CH2- (C-3′) position | 2.21, m | 34.03 | 2.29, m | 34.20 | 2.22, m | 34.05 |
-C=O | (173.43, 172.73) | (173.54, 172.41) | (173.59, 172.79) |
Gene | Sequence |
---|---|
LOR-F | 5′-GAGCTACGGAGGCGTCTCTA-3′ |
LOR-R | 5′-AGAGTAGCCGCAGACAGA-3′ |
FLG-F | 5′-GGTCTGGACGTTCAGGGTCT-3′ |
FLG-R | 5′-GGATGTGGTGTGGCTGTGAT-3′ |
TGM1-F | 5′-CCCCAAGAGACTAGCAGTGG-3′ |
TGM1-R | 5′-AGACCAGGCCATTCTTGATG-3′ |
GAPDH-F | 5′-GTGAAGGTCGGAGTCAACG-3′ |
GAPDH-R | 5′-TGAGGTCAATGAAGGGGTC-3′ |
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Jing, C.; Guo, J.; Li, Z.; Xu, X.; Wang, J.; Zhai, L.; Liu, J.; Sun, G.; Wang, F.; Xu, Y.; et al. Screening and Research on Skin Barrier Damage Protective Efficacy of Different Mannosylerythritol Lipids. Molecules 2022, 27, 4648. https://doi.org/10.3390/molecules27144648
Jing C, Guo J, Li Z, Xu X, Wang J, Zhai L, Liu J, Sun G, Wang F, Xu Y, et al. Screening and Research on Skin Barrier Damage Protective Efficacy of Different Mannosylerythritol Lipids. Molecules. 2022; 27(14):4648. https://doi.org/10.3390/molecules27144648
Chicago/Turabian StyleJing, Chenxu, Jiling Guo, Zhenzhuo Li, Xiaohao Xu, Jing Wang, Lu Zhai, Jianzeng Liu, Guang Sun, Fei Wang, Yangfen Xu, and et al. 2022. "Screening and Research on Skin Barrier Damage Protective Efficacy of Different Mannosylerythritol Lipids" Molecules 27, no. 14: 4648. https://doi.org/10.3390/molecules27144648