Moringa oleifera Leaves Protein Enhances Intestinal Permeability by Activating TLR4 Upstream Signaling and Disrupting Tight Junctions
Abstract
:1. Introduction
2. Results
2.1. Viability of Caco-2 Cells
2.2. Changes in the TEER of Monolayers of Cells
2.3. FITC-Dextran Cellular Transport
2.4. Effect of M. oleifera Leaves Protein on the mRNA Expression of Tight Junction Proteins in Caco-2 Cells
2.5. Effects of M. oleifera Leaves Protein on the Expression of Tight Junction Proteins in Caco-2 Cells
2.6. Effect of M. oleifera Leaves Protein Treatment on Occludin Protein Localization in Caco-2 Cells
2.7. Effects of M. oleifera Leaves Protein on Organ Indices in Mice
2.8. Serum Fluorescence Intensity in Mice
2.9. Effects of M. oleifera Leaves Protein on Tight Junction Protein Expression in the Small Intestinal Epithelium
3. Discussion
4. Materials and Methods
4.1. Materials and Chemicals
4.2. Caco-2 Cell Assay
4.2.1. Cell Viability Assay
4.2.2. Establishment and Grouping of the Caco-2 Cell Permeability Model
4.2.3. Evaluation of Cell Transepithelial Electrical Resistance (TEER)
4.2.4. Permeability Assays
4.2.5. Determination of the mRNA Expression of Tight Junction Proteins by Real-Time PCR (RT–PCR)
4.2.6. Immunoblot Analysis
4.2.7. Effect of M. oleifera Leaves Protein Treatment on Occludin Protein Localization in Caco-2 Cells
4.3. Animal Experiments
4.3.1. Study Subjects and Subgroups
4.3.2. Mouse Serum Fluorescence Intensity Assay
4.3.3. Assessment of Physiological Indicators in Mice
4.3.4. Western Blot Analysis
4.4. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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Genes | Forward | Reverse |
---|---|---|
Occludin | CTTCCAATGGCAAAGTGAATG | TACCACCGCTGCTGTAACGAG |
Claudin-1 | CCAGGTACGAATTTGGTCAGG | TGGTGTTGGGTAAGAGGTTGT |
ZO-1 | GAGCCTAATCTGACCTATGAACC | TGAGGACTCGTATCTGTATGTGG |
IL-8 | CCTGAACCTTCCAAAGATGGC | TTCACCAGGCAAGTCTCCTCA |
TLR4 | GTACCTGGGGAACAACCTCTT | GCAGCTTGACTAGACTCTCCA |
1RPL-19 | GAAGGTCAAAGGGAATGTGTTCA | CCTTGTCTGCCTTCAGCTTGT |
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Liu, X.; Xi, C.; Li, W.; Su, H.; Yang, H.; Bai, Z.; Tian, Y.; Song, S. Moringa oleifera Leaves Protein Enhances Intestinal Permeability by Activating TLR4 Upstream Signaling and Disrupting Tight Junctions. Int. J. Mol. Sci. 2023, 24, 16425. https://doi.org/10.3390/ijms242216425
Liu X, Xi C, Li W, Su H, Yang H, Bai Z, Tian Y, Song S. Moringa oleifera Leaves Protein Enhances Intestinal Permeability by Activating TLR4 Upstream Signaling and Disrupting Tight Junctions. International Journal of Molecular Sciences. 2023; 24(22):16425. https://doi.org/10.3390/ijms242216425
Chicago/Turabian StyleLiu, Xiaoxue, Chuyu Xi, Wenjie Li, Hairan Su, Hao Yang, Zhongbin Bai, Yang Tian, and Shuang Song. 2023. "Moringa oleifera Leaves Protein Enhances Intestinal Permeability by Activating TLR4 Upstream Signaling and Disrupting Tight Junctions" International Journal of Molecular Sciences 24, no. 22: 16425. https://doi.org/10.3390/ijms242216425