Citric Acid Promotes Immune Function by Modulating the Intestinal Barrier
Abstract
:1. Introduction
2. Results
2.1. Citric Acid Increases Body Weight and Stimulates Cell Growth by Increasing the Number of G2 Phase Cells in IEC-6 Cells of Mice
2.2. Improvement of Intestinal Structure by Citric Acid-Induced Increase in the Ratio of Villi to Crypts in Mouse Small Intestine
2.3. Citric Acid Enhances Growth of Common Probiotic Bacteria in the Cecum of Mice
2.4. Citric Acid Induced the Expression of Tight Junction Genes in Intestinal Tissue and IEC-6 Cells
2.5. Transcriptome and Metabolic Pathway Analysis of Citric Acid-Induced Serum Metabolism in Mice
2.6. Citric Acid Maintains Tight Junction Gene Expression in IEC-6 Cells during H9N2 Infection and Inhibits Viral Replication
3. Discussion
4. Materials and Methods
4.1. Cells, Animals and Reagents
4.2. Grouping of Mice and Plotting of Body Weight Curves
4.3. Cell Assay
4.3.1. Cell Aggregation and Growth Curve Graphing
4.3.2. Cell Cycle Assay
4.4. Detection of Morphological Structure of Mouse Intestine
4.5. Detection of Bacterial Flora in the Contents of Mouse Cecum
Copy number (copies·μL−1) = (6.02 × 1023 × (ng·μL−1 × 10−9)/(DNA length × 660)).
4.6. Intestinal Tight Junction Assay
4.6.1. Detection of Mouse Intestinal Tight Junction Genes
4.6.2. Cellular Tight Junction Protein Gene Expression Assay
4.7. Detection of Serum Metabolites in Mice Using Untargeted Metabolomics
4.8. Detection of Inflammatory Factors and Type I Interferon in IAV-Infected IEC-6 Cells
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Groups (g/L) | Villus Height to Crypt Depth Ratio |
---|---|
0 | 3.944 ± 0.2297 |
0.05 | 4.373 ± 0.2492 |
0.1 | 5.011 ± 0.2797 AA |
0.15 | 4.720 ± 0.1643 AA |
0.2 | 4.095 ± 0.6054 |
Bacterial Type | 0 g/L | 0.05 g/L | 0.1 g/L | 0.15 g/L | 0.2 g/L |
---|---|---|---|---|---|
Bifid. | 2.15 ± 0.03 | 2.09 ± 0.02 | 2.52 ± 0.01 AA | 2.34 ± 0.02 aa | 2.10 ± 0.07 |
Lac. | 2.38 ± 0.03 | 2.37 ± 0.02 | 2.50 ± 0.08 | 2.45 ± 0.06 | 2.47 ± 0.15 |
E. coli | 2.72 ± 0.01 | 2.72 ± 0.01 | 2.73 ± 0.01 | 2.71 ± 0.01 | 2.72 ± 0.01 |
Name | Impact | Details |
---|---|---|
Taurine and hypotaurine metabolism | 0.42857 | KEGG |
Arachidonic acid metabolism | 0.33292 | KEGG |
Cysteine and methionine metabolism | 0.10446 | KEGG |
Primary bile acid biosynthesis | 0.02239 | KEGG |
Purine metabolism | 0.01945 | KEGG |
Fatty acid biosynthesis | 0.01473 | KEGG |
Glutathione metabolism | 0.00709 | KEGG |
Steroid hormone biosynthesis | 0.00653 | KEGG |
Biosynthesis of unsaturated fatty acids | 0 | KEGG |
Phenylalanine metabolism | 0 | KEGG |
Fatty acid elongation | 0 | KEGG |
Aminoacyl-tRNA biosynthesis | 0 | KEGG |
Gene | GenBank No. | Direction | Sense and Antisense Primer (5′–3′) |
---|---|---|---|
Occludin | NM_029865.2 | Forward | GTACCCACCAGTGACCAACA |
Reverse | GTTGCTGGAGCTTAGCCTGT | ||
ZO-1 | XM_036152895.1 | Forward | CGAGGCATCATCCCAAATAAGAAC |
Reverse | TCCAGAAGTCTGCCCGATCAC | ||
Claudin-1 | NM_016674.4 | Forward | TGGTAATTGGCATCCTGCTG |
Reverse | CAGCCATCCACATCTTCTGC | ||
Bifid. | AB064848.1 | Forward | GATTCTGGCTCAGGATGAACGC |
Reverse | CTGATAGGACGAGACCCAT | ||
Lac. | OM349561.1 | Forward | AGCAGTAGGGATCTTCCA |
Reverse | ATTTCACCGCTACACATG | ||
E.coli | OU548744.1 | Forward | GGGAGTAAAGTTAATACCTTTGCTC |
Reverse | TTCCCGAAGGCACATTCT | ||
TNF-α | XM_021218152.1 | Forward | GAGGCACTCCCCCAAAAGAT |
Reverse | GAGGGAGGCCATTTGGGAAC | ||
IL-6 | NM_031168.2 | Forward | GTCCTTCCTACCCCAATTTCCA |
Reverse | TAACGCACTAGGTTTGCCGA | ||
IFN-β | NM_010510.2 | Forward | AACTCCACCAGCAGACAGTG |
Reverse | GGTACCTTTGCACCCTCCAG | ||
ISG15 | NM_015783.3 | Forward | CGGGAACAAGTCCACGAAG |
Reverse | CCCTCAGGCGCAAATGCT |
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Hu, P.; Yuan, M.; Guo, B.; Lin, J.; Yan, S.; Huang, H.; Chen, J.-L.; Wang, S.; Ma, Y. Citric Acid Promotes Immune Function by Modulating the Intestinal Barrier. Int. J. Mol. Sci. 2024, 25, 1239. https://doi.org/10.3390/ijms25021239
Hu P, Yuan M, Guo B, Lin J, Yan S, Huang H, Chen J-L, Wang S, Ma Y. Citric Acid Promotes Immune Function by Modulating the Intestinal Barrier. International Journal of Molecular Sciences. 2024; 25(2):1239. https://doi.org/10.3390/ijms25021239
Chicago/Turabian StyleHu, Pengcheng, Meng Yuan, Bolun Guo, Jiaqi Lin, Shihong Yan, Huiqing Huang, Ji-Long Chen, Song Wang, and Yanmei Ma. 2024. "Citric Acid Promotes Immune Function by Modulating the Intestinal Barrier" International Journal of Molecular Sciences 25, no. 2: 1239. https://doi.org/10.3390/ijms25021239