The PI3K/AKT cell signaling pathway is involved in regulation of osteoporosis

Background: Osteoporosis is a systemic skeletal disease with the high incidence, serious complications, financial burden, and heavily decrease in living quality. Methods: Proliferation of osteoblast was tested by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) method,...

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Published in:Journal of receptors and signal transduction 2015-11, Vol.35 (6), p.640-645
Main Authors: Xi, Jian-Cheng, Zang, Hai-Yu, Guo, Li-Xin, Xue, Hai-Bin, Liu, Xiang-Dong, Bai, Yi-Bing, Ma, Yuan-Zheng
Format: Article
Language:English
Subjects:
Akt
Animals
Apoptosis
Blotting, Western
bone formation
Calcium - metabolism
Cell Proliferation
Cells, Cultured
differentiation
Disease Models, Animal
Female
inhibitor
Osteoblasts - cytology
Osteoblasts - metabolism
osteoporosis
Osteoporosis - etiology
Osteoporosis - metabolism
Osteoporosis - pathology
Phosphatidylinositol 3-Kinases - genetics
Phosphatidylinositol 3-Kinases - metabolism
PI3K
proliferation
Proto-Oncogene Proteins c-akt - genetics
Proto-Oncogene Proteins c-akt - metabolism
RANK Ligand - metabolism
Rats
Rats, Wistar
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger - genetics
Signal Transduction
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Summary:Background: Osteoporosis is a systemic skeletal disease with the high incidence, serious complications, financial burden, and heavily decrease in living quality. Methods: Proliferation of osteoblast was tested by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) method, alkaline phosphatase (ALP) activity of osteoblasts was tested by ALP REAGENT, Calcium level was determined by a colorimetric assay, mRNA expression of phosphoinositide-3 kinase (PI3K), 3-phosphoinositide-dependent protein kinase 1 (PDK1), Akt, Caspase-3, Caspase-7, Caspase-9, osteocalcin (OCN), Osterix and Runx2 of osteoblasts was tested by RNA preparation and quantitative reverse transcription polymerase chain reaction (RT-PCR), and protein expression of phospho-PI3K, phospho-PDK1 and phospho-Akt was measured by Western Blot analysis. Results: In osteoporosis model rats, it found that mRNA expression of PI3K, PDK1 and Akt showed no changes while protein expression of phospho-PI3K, phospho-PDK1 and phospho-Akt in bone tissue was decreased dramatically. To further characterize the molecular mechanisms that regulate osteoporosis, we examined the contribution of the PI3K/Akt cell signaling pathway in cultured osteoblasts. It suggested that, the blockade of PI3K activation by LY294002, a specific inhibitor of the PI3K/Akt signaling pathway in osteoblasts, heavily inhibited cell proliferation, ALP activity, calcium accumulation, and mRNA expression of OCN, Osterix and Runx2. However, mRNA expression of Caspase-3 and Caspase-9 was promoted accordingly. Conclusion: The in vivo and in vitro studies indicated that the PI3K/Akt cell signaling pathway is involved in the inhibition of osteoporosis through promoting osteoblast proliferation, differentiation and bone formation.
ISSN:1079-9893
1532-4281
DOI:10.3109/10799893.2015.1041647