Osteoblast Demineralization Induced by Oxidized High-Density Lipoprotein via the Inflammatory Pathway Is Suppressed by Adiponectin
Abstract
:1. Introduction
2. Results
2.1. OxHDL Reduces the Formation of Mineral Nodules and Calcium Incorporation in HOBs
2.2. Effect of HDL, oxHDL, Adiponectin, and Co-Incubation of oxHDL with Adiponectin on the Expression of Bone Mineralization Markers (RUNX2, ALPL, COL1A2, BGLAP)
2.3. Effect of HDL, oxHDL, Adiponectin, and Co-Incubation of oxHDL with Adiponectin on the Expression of Osteoblastic Transcription Factor (STAT-3 and PPAR-α)
2.4. Effects of HDL, oxHDL, Adiponectin, and Co-Incubation of oxHDL with Adiponectin on the Expression of Inflammatory Markers (IL-6, TNF-α, RELA Proto-Oncogene NF-κβ Subunit (p65), and CREB1)
3. Discussion
4. Materials and Methods
4.1. Culturing and Maintenance of HOBs
4.2. Oxidation of HDL
4.3. Mineralization Detection via Alizarin Red Staining
4.4. Detection of Calcium Deposition Using the Calcium Colorimetric Assay
4.5. Quantification of Bone Mineralization (ALPL, COL1A2, BGLAP, and RUNX2), Osteoblast-Related Transcription Factors (STAT-3 and PPAR-α), and Inflammatory (IL-6, TNF-α, RELA Proto-Oncogene, NF-kB Subunit (P65), and CREB) Markers Using Quantitative Reverse Transcription-Polymerase Chain Reaction (RT-qPCR)
4.6. Statistical Analysis
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 Name | Gene Symbol | Gene Bank Accession Number | Primer Sequence |
---|---|---|---|
Alkaline phosphatase | ALPL | NM_000478.5 | F : CCAAGTACTGGCGAGACCAA |
R : TGTGGAGACACCCATCCCAT | |||
Bone gamma-carboxyglutamate protein | BGLAP | NM_199173.5 | F : TCACACTCCTCGCCCTATTG |
R : CTCTTCACTACCTCGCTGCC | |||
Runt-related transcription factor 2 | RUNX2 | NM_0010246303 | F : CCACCACTCACTACCACACC |
R : AAGGGTCCACTCTGGCTTTG | |||
Collagen type I alpha 2 chain | COL1A2 | NM_000089.3 | F : GGATGAGGAGACTGGCAACC |
R : TGCCCTCAGCAACAAGTTCA | |||
Interleukin 6 | IL-6 | NM_000600.4 | F : TGAGGAGACTTGCCTGGTGA |
R : GCATTTGTGGTTGGGTCAGG | |||
Tumor necrosis factor | TNF-α | NM_000594.3 | F: GGTCCTCTTCAAGGGCCAAG |
R: TCCTCCTCACAGGGCAATGA | |||
RELA proto-oncogene, NF-kB (p65) subunit | P65 | NM_021975.3 | F : AGGCTATCAGTCAGCGCATC |
R : TCCCCACGCTGCTCTTCTAT | |||
cAMP responsive element binding protein 1 | CREB1 | NM_004379.4 | F : CGAGAACCAGCAGAGTGGAG |
R : CGGTGGGAGCAGATGATGTT | |||
Signal transducer and activator of transcription 3 | STAT3 | NM_139276.2 | F : ACCATTGACCTGCCGATGTC |
R : GTGAGGGACTCAAACTGCCC | |||
Peroxisome proliferator activated receptor alpha | PPAR-α | NM_001001928.2 | F : AAGGCTGCAAGGGCTTCTTT |
R : ACATCCCGACAGAAAGGCAC | |||
Glyceraldehyde 3-phosphate dehydrogenase | GAPDH | NM_002046.6 | F : GGAGCGAGATCCCTCCAAAAT |
R : GGCTGTTGTCATACTTCTCATGG | |||
Hyperparathyroidism 1 | HRPT1 | NM_000194_2 | F : CCTGGCGTCGTGATTAGTGAT |
R : AGACGTTCAGTCCTGTCCATAA |
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Harun, N.H.; Froemming, G.R.A.; Mohd Ismail, A.; Nawawi, H.; Mokhtar, S.S.; Abd Muid, S. Osteoblast Demineralization Induced by Oxidized High-Density Lipoprotein via the Inflammatory Pathway Is Suppressed by Adiponectin. Int. J. Mol. Sci. 2022, 23, 14616. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232314616
Harun NH, Froemming GRA, Mohd Ismail A, Nawawi H, Mokhtar SS, Abd Muid S. Osteoblast Demineralization Induced by Oxidized High-Density Lipoprotein via the Inflammatory Pathway Is Suppressed by Adiponectin. International Journal of Molecular Sciences. 2022; 23(23):14616. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232314616
Chicago/Turabian StyleHarun, Noor Hanisa, Gabriele Ruth Anisah Froemming, Aletza Mohd Ismail, Hapizah Nawawi, Siti Shuhada Mokhtar, and Suhaila Abd Muid. 2022. "Osteoblast Demineralization Induced by Oxidized High-Density Lipoprotein via the Inflammatory Pathway Is Suppressed by Adiponectin" International Journal of Molecular Sciences 23, no. 23: 14616. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232314616