Increased Permeability of the Blood–Brain Barrier in a Diabetic Mouse Model (Leprdb/db Mice)
Abstract
:1. Introduction
2. Results
2.1. The Molecular Expression of BBB Markers in the Brain Homogenate of Leprdb/db Mice
2.2. Immunohistochemistry of ZO-1 in the Prefrontal Area of Leprdb/db and Control Mice
2.3. BBB Permeability Analysis Using Na+ Fluorescein
3. Discussion
Study Limitations
4. Materials and Methods
4.1. Animals
4.2. Experimental Design
4.3. Tissue Collection
4.4. BBB Permeability Experiment
4.4.1. Pharmacokinetic Studies
4.4.2. Spectrofluorometer Analysis of Na+ Fluorescein Permeability in the BBB
4.4.3. Pharmacokinetic Analysis
4.5. Quantification of mRNA Expression by Real-Time Polymerase Chain Reaction
4.5.1. CDNA Synthesis
4.5.2. Real-Time Polymerase Chain Reaction (RT-PCR)
4.6. Immunofluorescence
4.7. Confocal Microscopy
4.8. Western Blotting
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
BBB | Blood-Brain Barrier |
DM | Diabetes Mellitus |
T1DM | Type 1 Diabetes Mellitus |
T1DM | Type 1 Diabetes Mellitus |
ICAM | Intercellular Adhesion Molecule |
MMP | Matrix Metalloproteinases |
NaFL | Sodium Fluorescein |
RT-PCR | Real-Time Polymerase Chain Reaction |
VCAM | Vascular cell adhesion protein |
S100b | S100 calcium-binding protein B |
Leprdb/db | Leptin receptor mutant mice |
TJP-1 | Tight junction protein 1 (Zonula occludens-1) |
ZO-1 | Zonula occludens-1 |
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Groups (n = 3) | Mean | Std. Error of Mean |
---|---|---|
WT | 1.204 | ±0.1479 |
WT Treated | 1.613 | ±0.02765 |
Leprdb/db | 2.044 ** | ±0.01843 |
Leprdb/db Treated | 1.406 | ±0.1607 |
Groups (n = 3) | Mean | Std. Error of Mean |
---|---|---|
WT | 0.5210 | ±0.05255 |
WT Treated | 0.6077 | ±0.04840 |
Leprdb/db | 0.7883 | ±0.05990 |
Leprdb/db Treated | 0.5433 | ±0.06972 |
Gene | Name | NCBI ID | Forward | Reverse |
---|---|---|---|---|
ICAM-1 | intercellular adhesion molecule 1 | NM_010493.3 | GTG ATG GCA GCC TCT TAT GT | GGG CTT GTC CCT TGA GTT T |
VCAM-1 | vascular cell adhesion molecule 1 | NM_011693.3 | GAG GGA GAC ACC GTC ATT ATC | CGA GCC ATC CAC AGA CTT TA |
MMP | matrix metallopeptidase 9 | NM_013599.4 | TGC ACT GGG CTT AGA TCA TTC | TGC CGT CTA TGT CGT CTT TAT TC |
S100b | beta polypeptide, neural | NM_009115.3 | GCA AAT GAT GCT CCA GAA AGT AG | TGC ACA CTG TGG AAG AAG AG |
Gapdh | Glyceraldehyde-3-phosphate dehydrogenase isoform 1 | NM_001289726.1 | GTGGCAAAGTGGAGATTGTTG | CGTTGAATTTGCCGTGAGTG |
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Alshammari, M.A.; Alshehri, A.O.; Alqahtani, F.; Khan, M.R.; Bakhrebah, M.A.; Alasmari, F.; Alshammari, T.K.; Alsharari, S.D. Increased Permeability of the Blood–Brain Barrier in a Diabetic Mouse Model (Leprdb/db Mice). Int. J. Mol. Sci. 2024, 25, 7768. https://doi.org/10.3390/ijms25147768
Alshammari MA, Alshehri AO, Alqahtani F, Khan MR, Bakhrebah MA, Alasmari F, Alshammari TK, Alsharari SD. Increased Permeability of the Blood–Brain Barrier in a Diabetic Mouse Model (Leprdb/db Mice). International Journal of Molecular Sciences. 2024; 25(14):7768. https://doi.org/10.3390/ijms25147768
Chicago/Turabian StyleAlshammari, Musaad A., Abdulaziz O. Alshehri, Faleh Alqahtani, Mohammad R. Khan, Muhammed A. Bakhrebah, Fawaz Alasmari, Tahani K. Alshammari, and Shakir D. Alsharari. 2024. "Increased Permeability of the Blood–Brain Barrier in a Diabetic Mouse Model (Leprdb/db Mice)" International Journal of Molecular Sciences 25, no. 14: 7768. https://doi.org/10.3390/ijms25147768