NAD Supplement Alleviates Intestinal Barrier Injury Induced by Ethanol Via Protecting Epithelial Mitochondrial Function
Abstract
:1. Introduction
2. Methods
2.1. In Vivo Experiments
2.2. In Vitro Experiments
2.3. Histological Analysis
2.4. Biochemical Analysis
2.5. Measurements of Metabolites
2.6. RNA Isolation and Quantitative Real-Time PCR
2.7. Analysis of Mitochondrial DNA (mtDNA) Content
2.8. Mitochondrial Function Assay
2.9. Immunoprecipitation and Western Blot Analysis
2.10. Statistical Analysis
3. Results
3.1. NR Supplementation Alleviates Ethanol-Induced Intestinal Barrier Dysfunctions Both in In Vivo and In Vitro Experiments
3.2. NR Prevents Ethanol Induced Imbalance of NAD and Energy
3.3. NR Restores Ethanol-Induced Intestinal Mitochondrial Dysfunctions Both in In Vivo and In Vitro Experiments
3.4. NR Improves Mitochondrial Function and Intestinal Barrier Injuries in a SirT1-Dependent Manner
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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Gene | Species | Forward Primer | Reverse Primer |
---|---|---|---|
SirT1 | mouse | TGTGAAGTTACTGCAGGAGTGTAAA | GCATAGATACCGTCTCTTGATCTGAA |
PGC-1α | mouse | AAGTGTGGAACTCTCTGGAACTG | GGGTTATCTTGGTTGGCTTTATG |
ERRα | mouse | GGGGAGCATCGAGTACAGC | AGACGCACACCCTCCTTGA |
NRF1 | mouse | AGCACGGAGTGACCCAAAC | AGGATGTCCGAGTCATCATAAGA |
NRF2 | mouse | CTTTAGTCAGCGACAGAAGGAC | AGGCATCTTGTTTGGGAATGTG |
TFAM | mouse | AACACCCAGATGCAAAACTTTCA | GACTTGGAGTTAGCTGCTCTTT |
ATPase | mouse | GCCCGAGTGTATGGGACAG | GGACACACCAATAATGAGGTGC |
β-actin | mouse | GTGGTGGTGAAGCTGTAGCC | AGCCATGTACGTAGCCATCC |
SirT1 | human | TCTGGCATGTCCCACTATCA | TAGGCGGCTTGATGGTAATC |
PGC-1α | human | GCTTTCTGGGTGGACTCAAC | CTGCTAGCAAGTTTGCCTCA |
ERRα | human | TCCAGCTCCCACTCGCTGCC | ACACTCGTTGGAGGCCGGAC |
NRF1 | human | CCAGTTTAGTGGGTGGTAGG | CGGGAGCTTTCAAGACATTC |
NRF2 | human | GGCGCGTAGGTTTGTTCTAC | ACTCCAGCCATGACTAAAAGAGA |
TFAM | human | GTCACTGCCTCATCCACC | CCGCCCTATAAGCATCTT |
ATPase6 | human | TCAGCCTACTCATTCAAC | CTAGGATAGTCAGTAGAATTAGA |
MT-CO1 | human | CACACGAGCATATTTCAC | GTACGATGTCTAGTGATGA |
β-Globin | human | AAAGGTGCCCTTGAGGTTGTC | TGAAGGCTCATGGCAAGAAA |
β-actin | human | TCAAGAAAGGGTGTAACGCAACT | CGACAGGATGCAGAAGGAGAT |
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Li, W.; Zhou, Y.; Pang, N.; Hu, Q.; Li, Q.; Sun, Y.; Ding, Y.; Gu, Y.; Xiao, Y.; Gao, M.; et al. NAD Supplement Alleviates Intestinal Barrier Injury Induced by Ethanol Via Protecting Epithelial Mitochondrial Function. Nutrients 2023, 15, 174. https://doi.org/10.3390/nu15010174
Li W, Zhou Y, Pang N, Hu Q, Li Q, Sun Y, Ding Y, Gu Y, Xiao Y, Gao M, et al. NAD Supplement Alleviates Intestinal Barrier Injury Induced by Ethanol Via Protecting Epithelial Mitochondrial Function. Nutrients. 2023; 15(1):174. https://doi.org/10.3390/nu15010174
Chicago/Turabian StyleLi, Wenli, Yujia Zhou, Nengzhi Pang, Qianrong Hu, Qiuyan Li, Yan Sun, Yijie Ding, Yingying Gu, Ying Xiao, Mengqi Gao, and et al. 2023. "NAD Supplement Alleviates Intestinal Barrier Injury Induced by Ethanol Via Protecting Epithelial Mitochondrial Function" Nutrients 15, no. 1: 174. https://doi.org/10.3390/nu15010174