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Angiotensin II suppresses osteoblastic differentiation and mineralized nodule formation via AT1 receptor in ROS17/2.8 cells.

Nakai K, Kawato T, Morita T, Yamazaki Y, Tanaka H, Tonogi M, Oki H, Maeno M - Arch Med Sci (2015)

Bottom Line: Runx2, Msx2, and osteocalcin expression significantly decreased with Ang II compared to the control, whereas AJ18 expression significantly increased.Losartan blocked suppressive or stimulatory effects of Ang II on Runx2, Msx2, osteocalcin, and AJ18 expression.These results suggest that Ang II suppresses osteoblastic differentiation by altering the expression of osteogenesis-related transcription factors via the AT1 receptor and the function of osteogenesis in ROS17/2.8 cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Oral Health Sciences, Nihon University Graduate School of Dentistry, Tokyo, Japan.

ABSTRACT

Introduction: Angiotensin II (Ang II) not only regulates systemic blood pressure through a vasoconstrictive effect, but also promotes bone resorption. We recently reported that Ang II (10(-6) M) stimulated the production of matrix metalloproteinases via the AT1 receptor in osteoblastic ROS17/2.8 cells, but suppressed alkaline phosphatase activity. However, the roles of Ang II in osteoblastic differentiation and the function of osteogenesis in osteoblasts are unclear. Therefore, we examined the effect of Ang II on the expression of osteogenesis-related transcription factors and extracellular matrix (ECM) proteins, as well as mineralized nodule formation in ROS17/2.8 cells.

Material and methods: ROS17/2.8 cells were cultured with 0 (control) or 10(-6) M Ang II in the presence or absence of the AT1 receptor blocker losartan. Mineralized nodule formation was detected by Alizarin Red staining. Gene and protein expression levels of transcription factors and ECM proteins were determined using real-time PCR and Western blotting, respectively.

Results: Runx2, Msx2, and osteocalcin expression significantly decreased with Ang II compared to the control, whereas AJ18 expression significantly increased. Osterix, Dlx5, type I collagen, bone sialoprotein, and osteopontin expression was unaffected. Mineralized nodule formation and calcium content in mineralized nodules decreased with Ang II. Losartan blocked suppressive or stimulatory effects of Ang II on Runx2, Msx2, osteocalcin, and AJ18 expression.

Conclusions: These results suggest that Ang II suppresses osteoblastic differentiation by altering the expression of osteogenesis-related transcription factors via the AT1 receptor and the function of osteogenesis in ROS17/2.8 cells.

No MeSH data available.


Related in: MedlinePlus

Effect of Ang II on mRNA expression levels of ECM proteins. ROS17/2.8 cells were cultured with 0 (control) or 10–6 M Ang II for up to 7 days, and the mRNA expression levels of type I collagen (A), BSP (B), OPN (C), and OCN (D) on days 3, 5, and 7 of culture were determined using real-time PCREach bar indicates the mean ± SD from three independent experiments. **p < 0.01 (Ang II treatment vs. control).
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Figure 0004: Effect of Ang II on mRNA expression levels of ECM proteins. ROS17/2.8 cells were cultured with 0 (control) or 10–6 M Ang II for up to 7 days, and the mRNA expression levels of type I collagen (A), BSP (B), OPN (C), and OCN (D) on days 3, 5, and 7 of culture were determined using real-time PCREach bar indicates the mean ± SD from three independent experiments. **p < 0.01 (Ang II treatment vs. control).

Mentions: In both the presence and absence of 10–6 M Ang II, the mRNA expression level of type I collagen was highest on day 3 of culture, whereas the expression levels of BSP, OPN, and OCN were highest on day 5 (Figures 4 A–D). In the presence of 10–6 M Ang II, the mRNA expression level of OCN decreased significantly 0.71-fold compared to the control on day 5 of culture; the expression level was not affected on days 3 and 7 (Figure 4 D). The expression levels of type I collagen, BSP, and OPN were not affected by the addition of Ang II during the 7-day culture period (Figure 4 A–C).


Angiotensin II suppresses osteoblastic differentiation and mineralized nodule formation via AT1 receptor in ROS17/2.8 cells.

Nakai K, Kawato T, Morita T, Yamazaki Y, Tanaka H, Tonogi M, Oki H, Maeno M - Arch Med Sci (2015)

Effect of Ang II on mRNA expression levels of ECM proteins. ROS17/2.8 cells were cultured with 0 (control) or 10–6 M Ang II for up to 7 days, and the mRNA expression levels of type I collagen (A), BSP (B), OPN (C), and OCN (D) on days 3, 5, and 7 of culture were determined using real-time PCREach bar indicates the mean ± SD from three independent experiments. **p < 0.01 (Ang II treatment vs. control).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495158&req=5

Figure 0004: Effect of Ang II on mRNA expression levels of ECM proteins. ROS17/2.8 cells were cultured with 0 (control) or 10–6 M Ang II for up to 7 days, and the mRNA expression levels of type I collagen (A), BSP (B), OPN (C), and OCN (D) on days 3, 5, and 7 of culture were determined using real-time PCREach bar indicates the mean ± SD from three independent experiments. **p < 0.01 (Ang II treatment vs. control).
Mentions: In both the presence and absence of 10–6 M Ang II, the mRNA expression level of type I collagen was highest on day 3 of culture, whereas the expression levels of BSP, OPN, and OCN were highest on day 5 (Figures 4 A–D). In the presence of 10–6 M Ang II, the mRNA expression level of OCN decreased significantly 0.71-fold compared to the control on day 5 of culture; the expression level was not affected on days 3 and 7 (Figure 4 D). The expression levels of type I collagen, BSP, and OPN were not affected by the addition of Ang II during the 7-day culture period (Figure 4 A–C).

Bottom Line: Runx2, Msx2, and osteocalcin expression significantly decreased with Ang II compared to the control, whereas AJ18 expression significantly increased.Losartan blocked suppressive or stimulatory effects of Ang II on Runx2, Msx2, osteocalcin, and AJ18 expression.These results suggest that Ang II suppresses osteoblastic differentiation by altering the expression of osteogenesis-related transcription factors via the AT1 receptor and the function of osteogenesis in ROS17/2.8 cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Oral Health Sciences, Nihon University Graduate School of Dentistry, Tokyo, Japan.

ABSTRACT

Introduction: Angiotensin II (Ang II) not only regulates systemic blood pressure through a vasoconstrictive effect, but also promotes bone resorption. We recently reported that Ang II (10(-6) M) stimulated the production of matrix metalloproteinases via the AT1 receptor in osteoblastic ROS17/2.8 cells, but suppressed alkaline phosphatase activity. However, the roles of Ang II in osteoblastic differentiation and the function of osteogenesis in osteoblasts are unclear. Therefore, we examined the effect of Ang II on the expression of osteogenesis-related transcription factors and extracellular matrix (ECM) proteins, as well as mineralized nodule formation in ROS17/2.8 cells.

Material and methods: ROS17/2.8 cells were cultured with 0 (control) or 10(-6) M Ang II in the presence or absence of the AT1 receptor blocker losartan. Mineralized nodule formation was detected by Alizarin Red staining. Gene and protein expression levels of transcription factors and ECM proteins were determined using real-time PCR and Western blotting, respectively.

Results: Runx2, Msx2, and osteocalcin expression significantly decreased with Ang II compared to the control, whereas AJ18 expression significantly increased. Osterix, Dlx5, type I collagen, bone sialoprotein, and osteopontin expression was unaffected. Mineralized nodule formation and calcium content in mineralized nodules decreased with Ang II. Losartan blocked suppressive or stimulatory effects of Ang II on Runx2, Msx2, osteocalcin, and AJ18 expression.

Conclusions: These results suggest that Ang II suppresses osteoblastic differentiation by altering the expression of osteogenesis-related transcription factors via the AT1 receptor and the function of osteogenesis in ROS17/2.8 cells.

No MeSH data available.


Related in: MedlinePlus