Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and Treatments
2.2. Preparation of BSA-Conjugated Palmitic Acid (PA)
2.3. PA Treatment
2.4. Oil Red O Staining
2.5. Dihydroethidium Staining
2.6. RNA Extraction and Real-Time PCR Analysis
2.7. Immunofluorescence Staining
2.8. Statistical Analysis
3. Results
3.1. Validation of Palmitic Acid (PA) Treatment in the HBAs
3.2. PA Induces Oxidative Stress in the HBAs
3.3. PA Induces DNA Damage in the HBAs
3.4. PA Causes Cellular Senescence and SASP in the HBAs
3.5. PA Downregulates Glutamate Reuptake Transporter Expression in the HBAs
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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Human Gene | Forward Primer | Reverse Primer | Gene Accession No. |
---|---|---|---|
p16 | GATCCAGGTGGGTAGAGGGTC | CCCCTGCAAACTTCGTCCT | NM_058197.5 |
p21 | TGTCCGTCAGAACCCATGC | AAAGTCGAAGTTCCATCGCTC | NM_001374511.1 |
p53 | CAGCACATGACGGAGGTTGT | TCATCCAAATACTCCACACGC | NM_001407269.1 |
IL1α | CGCCAATGACTCAGAGGAAGA | AGGGCGTCATTCAGGATGAA | NM_000575.5 |
IL1β | AATCTGTACCTGTCCTGCGTGTT | TGGGTAATTTTTGGGATCTACACTCT | NM_000576.3 |
IL6 | CCGGGAACGAAAGAGAAGCT | GCGCTTGTGGAGAAGGAGTT | NM_000600.5 |
IL8 | CTTGGCAGCCTTCCTGATTT | TTCTTTAGCACTCCTTGGCAAAA | NM_001354840.3 |
TNF-α | CCCAGGGACCTCTCTCTAATCA | AGCTGCCCCTCAGCTTGAG | NM_000594.4 |
MMP3 | GACACCAGCATGAACCTTGTT | GGAACCGAGTCAGGTCTGTG | NM_002422.5 |
MMP13 | GGCTCCGAGAAATGCAGTCTTTCTT | ATCAAATGGGTAGAAGTCGCCATGC | NM_002427.4 |
MCP-1 | AATCAATGCCCCAGTCACCT | CTTCTTTGGGACACTTGCTGC | NM_002982.4 |
CXCL1 | GAAAGCTTGCCTCAATCCTG | CACCAGTGAGCTTCCTCCTC | NM_001511.4 |
CXCL10 | AACCTCCAGTCTCAGCACCATGAA | AGGTACAGCGTAAGGTTCTAGAGAG | NM_001565.4 |
NQO1 | ACTGATCGTACTGGCTCACTC | AGTTCATGGCATAGAGGTCCG | NM_001025434.2 |
SOD2 | GTTGGGGTTGGCTTGGTTTC | GTTCCTTGCAGTGGATCCTGA | NM_001322819.2 |
CAT | CTTTCTGTTGAAGATGCGGCG | AGTCCAGGAGGGGTACTTTCC | NM_001752.4 |
EAAT1 | TGCCCTGGGTCTAGTTGTCT | CCAGTCTCATGATGGCTTCGT | NM_004172.5 |
EAAT2 | CTTGGCATCTCCCATCCACC | GCTGGAGATGATTAGAGGGAGAA | NM_001252652.2 |
Lamin B1 | AAGGCGAAGAAGAGAGGTTGAAG | GCGGAATGAGAGATGCTAACACT | NM_001198557.2 |
Plin1 | CAGAAACAGCATCAGCGTTCC | CAGCAAATTCCGCAGTGTCTC | NM_001145311.2 |
Plin2 | TCACAGGGGTGATGGACAAG | TTTCTACGCCACTGCTCACG | NM_001122.4 |
GAPDH | GCCGTCTAGAAAAACCTGCC | ACCACCTGGTGCTCAGTGTA | NM_001357943.2 |
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Sivasubramanian, M.K.; Monteiro, R.; Jagadeesh, M.; Balasubramanian, P.; Subramanian, M. Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation. Nutrients 2024, 16, 2852. https://doi.org/10.3390/nu16172852
Sivasubramanian MK, Monteiro R, Jagadeesh M, Balasubramanian P, Subramanian M. Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation. Nutrients. 2024; 16(17):2852. https://doi.org/10.3390/nu16172852
Chicago/Turabian StyleSivasubramanian, Mahesh Kumar, Raisa Monteiro, Manoj Jagadeesh, Priya Balasubramanian, and Madhan Subramanian. 2024. "Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation" Nutrients 16, no. 17: 2852. https://doi.org/10.3390/nu16172852