Identification of kaempferol-regulated proteins in rat calvarial osteoblasts during mineralization by proteomics (Article, 2010) [WorldCat.org]
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Identification of kaempferol-regulated proteins in rat calvarial osteoblasts during mineralization by proteomics
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Identification of kaempferol-regulated proteins in rat calvarial osteoblasts during mineralization by proteomics

Author: Avinash Kumar Affiliation: Division of Endocrinology, Central Drug Research Institute, Chattar Manzil Palace, Lucknow, IndiaAnand K Singh Affiliation: Toxicogenomics and Predictive Toxicology Division, Indian Institute of Toxicology Research, Mahatma Gandhi Marg, Lucknow, IndiaAbnish K Gautam Affiliation: Division of Endocrinology, Central Drug Research Institute, Chattar Manzil Palace, Lucknow, IndiaDeepak Chandra Affiliation: Department of Biochemistry, Lucknow University, Lucknow, IndiaDivya Singh Affiliation: Division of Endocrinology, Central Drug Research Institute, Chattar Manzil Palace, Lucknow, IndiaAll authors
Edition/Format: Article Article : English
Publication:PROTEOMICS, v10 n9 (No. 9 May 2010): 1730-1739
Other Databases: WorldCatWorldCatWorldCat
Summary:
Kaempferol, a flavonoid, promotes osteoblast mineralization in vitro and bone formation in vivo; however, its mechanism of action is yet unknown. We adopted proteomic approach to identify the differential effect of kaempferol on rat primary calvarial osteoblasts during mineralization. The primary rat calvarial osteoblasts were treated with kaempferol (5.0 μM) for 9 days under mineralizing condition that resulted in significant increase in alkaline phosphatase activity and mineralization of the cells. Further, 2-D analysis of the kaempferol-treated osteoblast lysates revealed 18 differentially expressed proteins (nine upregulated and nine downregulated) on the basis of >/<2.0-fold as cut-off (p<0.01) that were then identified by MALDI-TOF MS. These included cytoskeletal proteins, intracellular signaling protein, chaperone, extracellular matrix protein, and proteins involved in glycolysis and cell-matrix interactions. Proteomics data were confirmed by Western blotting and quantitative real-time PCR by randomly selecting two upregulated and two downregulated proteins. Western blot analysis confirmed upregulation of HSP-70 and cytokeratin-14 levels, and downregulation of aldose reductase and caldesmon expression. We further demonstrated that kaempferol treatment inhibits aldose reductase activity in osteoblasts indicating an altered cellular metabolism by decelerating polyol pathway that was associated with the kaempferol-induced osteoblast mineralization. In conclusion, this is a first comprehensive study on the differential regulation of proteins by kaempferol in primary osteoblast, which would further help to elucidate the role of the identified proteins in the process of osteoblast mineralization.  Read more...
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Details

Document Type: Article
All Authors / Contributors: Avinash Kumar Affiliation: Division of Endocrinology, Central Drug Research Institute, Chattar Manzil Palace, Lucknow, India; Anand K Singh Affiliation: Toxicogenomics and Predictive Toxicology Division, Indian Institute of Toxicology Research, Mahatma Gandhi Marg, Lucknow, India; Abnish K Gautam Affiliation: Division of Endocrinology, Central Drug Research Institute, Chattar Manzil Palace, Lucknow, India; Deepak Chandra Affiliation: Department of Biochemistry, Lucknow University, Lucknow, India; Divya Singh Affiliation: Division of Endocrinology, Central Drug Research Institute, Chattar Manzil Palace, Lucknow, India; Bendangla Changkija Affiliation: Division of Endocrinology, Central Drug Research Institute, Chattar Manzil Palace, Lucknow, India; Mahendra Pratap Singh Affiliation: Toxicogenomics and Predictive Toxicology Division, Indian Institute of Toxicology Research, Mahatma Gandhi Marg, Lucknow, India; Ritu Trivedi Affiliation: Division of Endocrinology, Central Drug Research Institute, Chattar Manzil Palace, Lucknow, India
ISSN:1615-9853
Language Note: English
Unique Identifier: 5151266448
Notes: Number of Figures: 5
Number of Tables: 1
Number of References: 52
Awards:

Abstract:

Kaempferol, a flavonoid, promotes osteoblast mineralization in vitro and bone formation in vivo; however, its mechanism of action is yet unknown. We adopted proteomic approach to identify the differential effect of kaempferol on rat primary calvarial osteoblasts during mineralization. The primary rat calvarial osteoblasts were treated with kaempferol (5.0 μM) for 9 days under mineralizing condition that resulted in significant increase in alkaline phosphatase activity and mineralization of the cells. Further, 2-D analysis of the kaempferol-treated osteoblast lysates revealed 18 differentially expressed proteins (nine upregulated and nine downregulated) on the basis of >/<2.0-fold as cut-off (p<0.01) that were then identified by MALDI-TOF MS. These included cytoskeletal proteins, intracellular signaling protein, chaperone, extracellular matrix protein, and proteins involved in glycolysis and cell-matrix interactions. Proteomics data were confirmed by Western blotting and quantitative real-time PCR by randomly selecting two upregulated and two downregulated proteins. Western blot analysis confirmed upregulation of HSP-70 and cytokeratin-14 levels, and downregulation of aldose reductase and caldesmon expression. We further demonstrated that kaempferol treatment inhibits aldose reductase activity in osteoblasts indicating an altered cellular metabolism by decelerating polyol pathway that was associated with the kaempferol-induced osteoblast mineralization. In conclusion, this is a first comprehensive study on the differential regulation of proteins by kaempferol in primary osteoblast, which would further help to elucidate the role of the identified proteins in the process of osteoblast mineralization.

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