Intestinal Region-Dependent Impact of NFκB-Nrf Crosstalk in Myenteric Neurons and Adjacent Muscle Cells in Type 1 Diabetic Rats
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animal Model
2.2. Tissue Handling
2.3. RNA Extraction, Reverse Transcription and Quantitative PCR
2.4. Primers
2.5. Measurement of Tissue NFκB and Nrf2 Concentrations
2.6. Bradford Protein Micromethod
2.7. Fluorescent Immunohistochemistry
2.8. Quantitative Post-Embedding Immunohistochemistry
2.9. Statistical Analysis
3. Results
3.1. Disease Characteristics of Type 1 Diabetic Rats
3.2. Diabetic Alterations of Intestinal NFκB Expression
3.2.1. Quantitative Changes in NFκB p65 mRNA Expression
3.2.2. NFκB p65 Protein Level in Smooth Muscle/Myenteric Plexus Homogenates
3.2.3. Proportion of NFκB p65-Immunoreactive Myenteric Neurons
3.2.4. Quantitative Distribution of NFκB p65-Labelling Gold Particles in Myenteric Ganglia and Intestinal Smooth Muscle
3.3. Diabetic Alterations of Intestinal Nrf2 Expression
3.3.1. Quantitative Changes in Nrf2 mRNA Expression
3.3.2. Nrf2 Protein Level in Smooth Muscle/Myenteric Plexus Homogenates
3.3.3. Quantitative Evaluation of Nrf2 Immunohistochemistry in Myenteric Ganglia and Intestinal Smooth Muscle
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Primers (5′ → 3′) |
---|---|
NFκB (NM_199267) | CCGGAACTCTGGGAGCTGCC AGGCTAGGGTCAGCGTATGG |
NRF2 (XM_006234397) | GCAACTCCAGAAGGAACAGG GGAATGTCTCTGCCAAAAGC |
Body Weight (g) | Blood Glucose Concentration (mmol/L) | |||
---|---|---|---|---|
Initial | Final | Initial | Final (Average) | |
Controls (n = 14) | 256.7 ± 17.68 | 467 ± 16.13 **** | 5.96 ± 0.26 | 5.92 ± 0.11 |
Diabetics (n = 14) | 223.9 ± 7.54 | 320.9 ± 14.73 | 6.09 ± 0.37 | 29.46 ± 0.87 ****,oooo |
Insulin-treated diabetics (n = 13) | 233.7 ± 9.44 | 463.1 ± 14.87 **** | 5.86 ± 0.35 | 12.98 ± 1.5 *,o |
Myenteric Neurons | Smooth Muscle Cells | |||
---|---|---|---|---|
Perikarya | Nuclei | Cytoplasm | Nuclei | |
Controls | 1.12 ± 0.16 | 0.95 ± 0.06 | 0.67 ± 0.04 | 1.27 ± 0.11 |
Diabetics | 1.43 ± 0.09 ** | 1.28 ± 0.08 * | 0.92 ± 0.05 *** | 1.35 ± 0.10 |
Insulin-treated diabetics | 1.32 ± 0.09 * | 1.16 ± 0.07 | 0.75 ± 0.04 ° | 0.97 ± 0.08 ° |
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Barta, B.P.; Onhausz, B.; Egyed-Kolumbán, A.; AL Doghmi, A.; Balázs, J.; Szalai, Z.; Ferencz, Á.; Hermesz, E.; Bagyánszki, M.; Bódi, N. Intestinal Region-Dependent Impact of NFκB-Nrf Crosstalk in Myenteric Neurons and Adjacent Muscle Cells in Type 1 Diabetic Rats. Biomedicines 2024, 12, 2347. https://doi.org/10.3390/biomedicines12102347
Barta BP, Onhausz B, Egyed-Kolumbán A, AL Doghmi A, Balázs J, Szalai Z, Ferencz Á, Hermesz E, Bagyánszki M, Bódi N. Intestinal Region-Dependent Impact of NFκB-Nrf Crosstalk in Myenteric Neurons and Adjacent Muscle Cells in Type 1 Diabetic Rats. Biomedicines. 2024; 12(10):2347. https://doi.org/10.3390/biomedicines12102347
Chicago/Turabian StyleBarta, Bence Pál, Benita Onhausz, Abigél Egyed-Kolumbán, Afnan AL Doghmi, János Balázs, Zita Szalai, Ágnes Ferencz, Edit Hermesz, Mária Bagyánszki, and Nikolett Bódi. 2024. "Intestinal Region-Dependent Impact of NFκB-Nrf Crosstalk in Myenteric Neurons and Adjacent Muscle Cells in Type 1 Diabetic Rats" Biomedicines 12, no. 10: 2347. https://doi.org/10.3390/biomedicines12102347