Crude Blueberry Phenolic Extracts Improve Gut Barrier Integrity and Exert Anti-Inflammatory and Antimicrobial Activity in an In Vitro Weaning Stress Model
Abstract
:1. Introduction
2. Materials and Methods
2.1. Crude Phenolic Extraction of Blueberries
2.2. Total Monomeric Anthocyanin Content Determination
2.3. Characterization of Blueberry Phenolic Extract
2.4. Cell Culture and Treatments
2.5. Cytotoxicity Assay
2.6. Catalase Activity Assay
2.7. Paracellular Permeability Assay
2.8. RNA Extraction and Real-Time Polymerase Chain Reaction (RT-PCR) Analysis
2.9. Western Blot Analyses of Tight Junction Proteins
2.10. Minimum Inhibitory Concentration Assay
2.11. Statistical Analysis
3. Results
3.1. Determination of BPE Phenolic Compounds
3.2. BPE Is Non-Cytotoxic to IPEC-J2 Cells at Low Concentrations
3.3. BPE Pretreatment Conserves Antioxidant Status after LPS Challenge at Low Concentrations
3.4. BPE Pretreatment Reduces Paracellular Permeability after LPS Challenge
3.5. BPE Pretreatment Decreases Expression of Inflammatory Markers and Upregulates Expression of Tight Junction Proteins
3.6. Pretreatment with BPE Is Able to Preserve the Expression of Tight Junction Protein OCLN
3.7. BPE Exhibits Antimicrobial Activity against Common PWD Pathogens
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Genes | Genes Accession Number | Primer Sequences |
---|---|---|
TNF-α | P23563 | F: 5′-ATGGATGGGTGGATGAGAAA-3′ R: 5′-TGGAAACTGTTGGGGAGAAG-3′ |
IL-8 | P26894 | F: 5′-CACCTGTCTGTCCACGTTGT-3′ R: 5′-AGAGGTCTGCCTGGACCCCA-3′ |
OCLN | A0A287AL69 | F: 5′-GAGAGAGTGGACAGCCCCAT-3′ R: 5′-TGCTGCTGTAATGAGGCTGC-3′ |
IL-1β | P26889 | F: 5′-CCAAAGAGGGACATGGAGAA-3′ R: 5′-GGGCTTTTGTTCTGCTTGAG-3′ |
IL-6 | P26893 | F: 5′-TCTGGGTTCAATCAGGAGACCTGC-3′ R: 5′-TGCACGGCCTCGACATTTCCC-3′ |
CLDN-1 | A0A287A1F1 | F: 5′-TTTCCTCAATACAGGAGGGAAGC-3′ R: 5′-CCCTCTCCCCACATTCGAG-3′ |
β-actin | Q80X90 | F: 5′ AGCCATGTACGTAGCCATCC-3′ R: 5′-CTCTCAGCTGTGGTGGTGAA-3′ |
Metabolite | Area Under Curve (±) SE |
---|---|
Hydroxycinnamic acids | |
3-Hydroxycinnamic acid | 15,033 ± 188 |
Caffeic acid | 91,956 ± 215 |
Flavonoid-3-O-glycosides | |
Isoquercitrin | 4663 ± 806 |
Quercetin-3-O-xyloside | 151,812 ± 3697 |
Isorhamnetin-3-O-beta-D-Glucoside | 328,457 ± 4587 |
Cyanidin-3-O-alpha-arabinopyranoside * | 1,558,051 ± 3961 |
Quercetin-3-O-xyloside | 7692 ± 295 |
5,7-dihydroxy-2-[4-hydroxy-3-[(2S,3R,4S,5R)-3,4,5-trihydroxyoxan-2-yl]oxyphenyl]-3-methoxychromen-4-one | 119,690 ± 1817 |
Cyanidin-3-O-alpha-arabinopyranoside * | 5358 ± 278 |
Peonidin-3-O-alpha-arabinopyranoside * | 70,535 ± 846 |
Quercetin-3-O-glucosyl-6′′-acetate | 80,661 ± 993 |
Peonidin-3-O-alpha-arabinoside * | 10,584 ± 465 |
Myricetin-3-Xyloside | 9074 ± 485 |
Isoquercetin | 123,995 ± 2503 |
Quercetin-3-Arabinoside | 29,406 ± 988 |
Quercetin 3-O-malonylglucoside | 8552 ± 223 |
Kaempferol-3-glucoside | 5162 ± 49 |
5,7-dihydroxy-2-(4-hydroxy-3-methoxyphenyl)-3-[3,4,5-trihydroxy-6-[[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxymethyl]oxan-2-yl]oxychromen-4-one | 4754 ± 266 |
Quercetin-3-O-alpha-L-rhamnopyranoside | 32,385 ± 1379 |
Isorhamnetin-3-O-beta-D-Glucoside | 6642 ± 94 |
Quercetin-3-O-glucosyl-6′′-acetate | 5422 ± 157 |
Quercetin-3-O-glucosyl-6′′-acetate | 9679 ± 443 |
2-(3,4-dihydroxyphenyl)-5,8-dihydroxy-7-methoxy-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxychromen-4-one | 6220 ± 303 |
Flavonoid-3-O-glucuronides | |
Quercetin 3-O-glucuronide | 66,122 ± 1386 |
Kaempferol 3-glucuronide | 6642 ± 94 |
Flavonoid-7-O-glycosides | |
Nepetin-7-glucoside | 3270 ± 430 |
Plantaginin | 240,153 ± 8929 |
NCGC00380911-01!2-(3,4-dihydroxyphenyl)-3,5-dihydroxy-8-methoxy-7-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxychromen-4-one | 18,211 ± 321 |
Anthocyanidin-3-O-glycosides | |
Delphinidin 3-glucoside * | 3557 ± 588 |
Petunidin-3-O-beta-glucoside * | 2141 ± 393 |
Delphinidin-3-O-beta-glucopyranoside * | 370,983 ± 6428 |
Peonidin-3-o-beta-d-glucopyranoside * | 148,567 ± 3296 |
Flavonols | |
Quercetin | 31,625 ± 245 |
Kaempferol | 22,206 ± 854 |
Isorhamnetin | 25,146 ± 999 |
Myricetin | 9599 ± 225 |
Isorhamnetin | 6573 ± 357 |
Limocitrin | 11,597 ± 481 |
Flavones | |
Luteolin | 18,168 ± 243 |
Flavanones | |
2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-chromen-4-one | 5354 ± 223 |
Catechins | |
Epicatechin | 20,234 ± 562 |
Epigallocatechins | |
Epigallocatechin | 4253 ± 217 |
Quinic acids and derivatives | |
Chlorogenic acid | 3354 ± 254 |
Biflavonoids and polyflavonoids | |
Procyanidin B1 | 25,277 ± 688 |
Phenolic glycosides | |
(E)-3-[4-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]prop-2-enoic acid | 5894 ± 395 |
6-O-methylated flavonoids | |
4′,5,7-trihydroxy-3,6-dimethoxyflavone | 14,776 ± 448 |
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Nathan, V.B.; Eckrote, S.; Li, S.; Reddivari, L. Crude Blueberry Phenolic Extracts Improve Gut Barrier Integrity and Exert Anti-Inflammatory and Antimicrobial Activity in an In Vitro Weaning Stress Model. Antioxidants 2024, 13, 1044. https://doi.org/10.3390/antiox13091044
Nathan VB, Eckrote S, Li S, Reddivari L. Crude Blueberry Phenolic Extracts Improve Gut Barrier Integrity and Exert Anti-Inflammatory and Antimicrobial Activity in an In Vitro Weaning Stress Model. Antioxidants. 2024; 13(9):1044. https://doi.org/10.3390/antiox13091044
Chicago/Turabian StyleNathan, Vignesh B., Sarah Eckrote, Shiyu Li, and Lavanya Reddivari. 2024. "Crude Blueberry Phenolic Extracts Improve Gut Barrier Integrity and Exert Anti-Inflammatory and Antimicrobial Activity in an In Vitro Weaning Stress Model" Antioxidants 13, no. 9: 1044. https://doi.org/10.3390/antiox13091044