In Vitro Fecal Fermentation of Euphorbia humifusa-Derived Polysaccharides and Their Protective Effect against Ulcerative Colitis in Mice
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Extraction and Purification of EHPs
2.3. Structural Characterization of EHPs
2.3.1. Chemical Composition
2.3.2. Molecular Weight (Mw) Distribution
2.3.3. Monosaccharide Composition
2.3.4. Fourier Transform Infrared (FTIR) Spectroscopic Analysis
2.4. In Vitro Absorption of EHPs
2.4.1. Fluorescent Labeling of Polysaccharides
2.4.2. Caco-2 Cell-Based Intestinal Absorption Model
2.4.3. EHP Uptake by Caco-2 Cells
2.5. In Vitro Fermentation of EHPs
2.5.1. Fermentation
2.5.2. Determination of SCFA Concentration
2.5.3. DNA Extraction and Sequence Analysis
2.6. Animal Studies
2.6.1. Experimental Design
2.6.2. qRT-PCR
2.7. Statistical Data Analysis
3. Results
3.1. Structural Characterization of EHPs
3.2. In Vitro Absorption Characteristics of Polysaccharides
3.3. In Vitro Fermentation of EHPs
3.3.1. Changes in Intestinal Microecology
3.3.2. Changes in SCFA Content
3.4. Therapeutic Effects of EHP Treatment in DSS-Induced Colitis Mice
3.4.1. EHP Supplementation Ameliorated Colitis Symptoms
3.4.2. EHP Treatment Inhibited Colonic Inflammation
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
Papp | Apparent permeability coefficient |
Ara | Arabinose |
DSS | Dextran sulfate sodium |
EHPs | Euphorbia humifusa-derived polysaccharides |
FTIR | Fourier transform infrared spectroscopy |
FITC | Fluorescein isothiocyanate |
HPSEC | High-performance size exclusion chromatography |
H&E | Hematoxylin-eosin |
Gal | Galactose |
Glu | Glucose |
GluA | Glucuronic acid |
Man | Mannose |
SCFAs | Short-chain fatty acids |
TEER | Transepithelial electrical resistance |
UC | Ulcerative colitis |
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Genes | Forward Primer | Reverse Primer |
---|---|---|
IL-6 | 5′-AGCGATGATGCACTGTCAGA-3′ | 5′-GGAACTCCAGAAGACCAGAGC-3′ |
IL-17 | 5′-TTCTTTCAAACAAAGGACCAGC-3′ | 5′-GCAACCCAAGTAACCCTTAAAG-3′ |
IL-10 | 5′-GACTTCACCATGGAACCCGT-3′ | 5′-GGAGACTGCCCATTCTCGAC-3′ |
Items | Content |
---|---|
Composition (w%) | |
Carbohydrate | 75.30 ± 0.04 |
Protein | 2.02 ± 0.41 |
Monosaccharide composition (molar ratio %) | |
Mannose (Man) | 6.12 |
Rhamnose (Rha) | 1.47 |
Glucuronic acid (GluA) | 8.30 |
Galacturonic acid (GalA) | 0.77 |
Glucose (Glu) | 21.71 |
Galactose (Gal) | 55.34 |
Arabinose (Ara) | 6.30 |
Sample | Time (h) | SCFAs(mmol/L) | ||||||
---|---|---|---|---|---|---|---|---|
Acetic Acid | Propionic Acid | Butyric Acid | Isobutyric Acid | Valeric Acid | Isovaleric Acid | Total | ||
Control | 0 | ND | ND | ND | ND | ND | ND | ND |
4 | 1.21 ± 0.21 f | 1.51 ± 0.17 d | 0.18 ± 0.01 e | 0.07 ± 0.00 c | 0.07 ± 0.00 d | 0.05 ± 0.00 b | 3.09 ± 0.11 f | |
8 | 2.61 ± 0.09 e | 2.43 ± 0.29 cd | 1.53 ± 0.16 cd | 0.10 ± 0.01 c | 0.11 ± 0.01 cd | 0.05 ± 0.00 b | 6.63 ± 0.10 de | |
12 | 5.62 ± 0.26 c | 6.20 ± 0.35 b | 5.80 ± 0.67 b | 1.70 ± 0.18 b | 0.25 ± 0.01 bc | 0.16 ± 0.01 b | 19.73 ± 1.84 c | |
24 | 6.10 ± 0.46 c | 7.01 ± 0.30 b | 7.34 ± 0.27 a | 3.46 ± 0.39 a | 0.41 ± 0.02 b | 2.14 ± 0.12 a | 26.46 ± 2.55 b | |
EHPs | 0 | ND | ND | ND | ND | ND | ND | ND |
4 | 1.64 ± 0.08 ef | 1.85 ± 0.09 d | 0.24 ± 0.01 de | 0.07 ± 0.01 c | 0.06 ± 0.00 d | 0.04 ± 0.00 b | 3.90 ± 0.67 ef | |
8 | 3.70 ± 0.42 d | 3.48 ± 0.35 c | 2.12 ± 0.42 c | 0.13 ± 0.01 c | 0.14 ± 0.01 cd | 0.07 ± 0.00 b | 9.64 ± 0.35 d | |
12 | 7. 50 ± 0.50 b | 7.20 ± 0.24 b | 5.43 ± 0.46 b | 1.81 ± 0.17 b | 0.35 ± 0.01 bc | 0.22 ± 0.02 b | 22.51 ± 1.81 c | |
24 | 8.82 ± 0.32 a | 9.92 ± 0.98 a | 7.62 ± 0.39 a | 3.24 ± 0.40 a | 0.90 ± 0.05 a | 2.25 ± 0.19 a | 32.75 ± 1.10 a |
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Xiang, N.; Zhao, J.; Chang, S.; Li, S.; Liu, S.; Wang, C. In Vitro Fecal Fermentation of Euphorbia humifusa-Derived Polysaccharides and Their Protective Effect against Ulcerative Colitis in Mice. Foods 2023, 12, 751. https://doi.org/10.3390/foods12040751
Xiang N, Zhao J, Chang S, Li S, Liu S, Wang C. In Vitro Fecal Fermentation of Euphorbia humifusa-Derived Polysaccharides and Their Protective Effect against Ulcerative Colitis in Mice. Foods. 2023; 12(4):751. https://doi.org/10.3390/foods12040751
Chicago/Turabian StyleXiang, Ning, Jianbo Zhao, Siqiao Chang, Shasha Li, Shuwen Liu, and Chan Wang. 2023. "In Vitro Fecal Fermentation of Euphorbia humifusa-Derived Polysaccharides and Their Protective Effect against Ulcerative Colitis in Mice" Foods 12, no. 4: 751. https://doi.org/10.3390/foods12040751