Plumbagin modulates RANKL-induced osteoclast and osteoblast differentiation
Background: Bone remodeling is a balanced, dynamic function of two counteractive processes: osteoblasts-mediated bone formation and osteoclasts-dependent bone resorption. An imbalance between these two processes causes bone diseases. Herein, the effects and molecular modus operandi of plumbagin- a b...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-05-01
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| Series: | Phytomedicine Plus |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667031323000258 |
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| author | Soni Ghumnani Vitthal T. Barvkar Suhas T. Mhaske Bhaskar Saha Richa Ashma |
| author_facet | Soni Ghumnani Vitthal T. Barvkar Suhas T. Mhaske Bhaskar Saha Richa Ashma |
| author_sort | Soni Ghumnani |
| collection | DOAJ |
| description | Background: Bone remodeling is a balanced, dynamic function of two counteractive processes: osteoblasts-mediated bone formation and osteoclasts-dependent bone resorption. An imbalance between these two processes causes bone diseases. Herein, the effects and molecular modus operandi of plumbagin- a bioactive phytometabolite from Plumbago sp. have been investigated on osteoblast and osteoclast differentiation. Material and Methods: Cell viability assay and Alizarin red-S staining assessed the plumbagin effects on pre-osteoblastic MC3T3-E1 cell viability and differentiation. Alkaline phosphatase activity, the osteoblast-specific genes, and protein expression were investigated by colorimetry, quantitative-PCR and immunoblotting, respectively. TRAP assay and osteoclast-specific proteins' expression assessed plumbagin's effect on osteoclastogenesis. During plumbagin-driven osteoclastogenesis, we used paired-end RNA-sequencing data to identify the key up-/down-regulated DEGs, validation by qPCR, and bioinformatics-based pathway analysis. Results: Plumbagin at a very low dose (0.25 µM) significantly promoted osteoblast differentiation, as evident from alkaline phosphatase, matrix mineralization, and enhanced expression of osteoblast-specific markers, including Runx2 and its target genes. The reduced TRAP activity, osteoclast-specific protein expression, and RNA sequencing suggested RANKL-induced osteoclastogenesis inhibition by plumbagin at 2 µM.. Conclusion: Plumbagin significantly enhances osteoblast differentiation by enhanced matrix mineralization and osteoblast-specific gene expression but suppresses the RANKL-induced osteoclastogenesis by regulating key osteoclast-specific DEGs and protein expression. Plumbagin can thus significantly influence bone biology and affect bone diseases. |
| first_indexed | 2024-04-09T13:21:41Z |
| format | Article |
| id | doaj.art-07e29acc26a34380af2440c555798274 |
| institution | Directory Open Access Journal |
| issn | 2667-0313 |
| language | English |
| last_indexed | 2024-04-09T13:21:41Z |
| publishDate | 2023-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Phytomedicine Plus |
| spelling | doaj.art-07e29acc26a34380af2440c5557982742023-05-11T04:24:52ZengElsevierPhytomedicine Plus2667-03132023-05-0132100429Plumbagin modulates RANKL-induced osteoclast and osteoblast differentiationSoni Ghumnani0Vitthal T. Barvkar1Suhas T. Mhaske2Bhaskar Saha3Richa Ashma4Department of Zoology, Savitribai Phule Pune University, Pune, 411 007, IndiaDepartment of Botany, Savitribai Phule Pune University, Pune, 411 007, IndiaRodent Experimentation Facility, ICMR-National Animal Resource Facility for Biomedical Research, Hyderabad, IndiaDepartment of Zoology, Savitribai Phule Pune University, Pune, 411 007, India; Department of Botany, Savitribai Phule Pune University, Pune, 411 007, India; Rodent Experimentation Facility, ICMR-National Animal Resource Facility for Biomedical Research, Hyderabad, India; National Centre For Cell Science Complex, University of Pune Campus, Pune University Rd, Ganeshkhind, Pune, Maharashtra 411007, IndiaDepartment of Zoology, Savitribai Phule Pune University, Pune, 411 007, India; Corresponding author at: Department of Zoology, Savitribai Phule Pune University, Pune- 411007. Maharashtra, India.Background: Bone remodeling is a balanced, dynamic function of two counteractive processes: osteoblasts-mediated bone formation and osteoclasts-dependent bone resorption. An imbalance between these two processes causes bone diseases. Herein, the effects and molecular modus operandi of plumbagin- a bioactive phytometabolite from Plumbago sp. have been investigated on osteoblast and osteoclast differentiation. Material and Methods: Cell viability assay and Alizarin red-S staining assessed the plumbagin effects on pre-osteoblastic MC3T3-E1 cell viability and differentiation. Alkaline phosphatase activity, the osteoblast-specific genes, and protein expression were investigated by colorimetry, quantitative-PCR and immunoblotting, respectively. TRAP assay and osteoclast-specific proteins' expression assessed plumbagin's effect on osteoclastogenesis. During plumbagin-driven osteoclastogenesis, we used paired-end RNA-sequencing data to identify the key up-/down-regulated DEGs, validation by qPCR, and bioinformatics-based pathway analysis. Results: Plumbagin at a very low dose (0.25 µM) significantly promoted osteoblast differentiation, as evident from alkaline phosphatase, matrix mineralization, and enhanced expression of osteoblast-specific markers, including Runx2 and its target genes. The reduced TRAP activity, osteoclast-specific protein expression, and RNA sequencing suggested RANKL-induced osteoclastogenesis inhibition by plumbagin at 2 µM.. Conclusion: Plumbagin significantly enhances osteoblast differentiation by enhanced matrix mineralization and osteoblast-specific gene expression but suppresses the RANKL-induced osteoclastogenesis by regulating key osteoclast-specific DEGs and protein expression. Plumbagin can thus significantly influence bone biology and affect bone diseases.http://www.sciencedirect.com/science/article/pii/S2667031323000258Bone cell differentiationPlumbaginMC3T3-E1 cell lineNFATc1RNA sequencing |
| spellingShingle | Soni Ghumnani Vitthal T. Barvkar Suhas T. Mhaske Bhaskar Saha Richa Ashma Plumbagin modulates RANKL-induced osteoclast and osteoblast differentiation Phytomedicine Plus Bone cell differentiation Plumbagin MC3T3-E1 cell line NFATc1 RNA sequencing |
| title | Plumbagin modulates RANKL-induced osteoclast and osteoblast differentiation |
| title_full | Plumbagin modulates RANKL-induced osteoclast and osteoblast differentiation |
| title_fullStr | Plumbagin modulates RANKL-induced osteoclast and osteoblast differentiation |
| title_full_unstemmed | Plumbagin modulates RANKL-induced osteoclast and osteoblast differentiation |
| title_short | Plumbagin modulates RANKL-induced osteoclast and osteoblast differentiation |
| title_sort | plumbagin modulates rankl induced osteoclast and osteoblast differentiation |
| topic | Bone cell differentiation Plumbagin MC3T3-E1 cell line NFATc1 RNA sequencing |
| url | http://www.sciencedirect.com/science/article/pii/S2667031323000258 |
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