Effect of cytoskeleton integrity on the expression of COX-2 gene in osteoblasts induced by fluid shear stress

doi:10.3877 / cma.j.issn.1674-1366.2009-04-009

Abstract

Abstract:

Objective

To study the effect of cytoskeleton integrity on the expression of COX-2 gene in osteoblasts induced by fluid shear stress.

Methods

BALB/c mouse primary osteoblasts were isolated by enzyme digestion technique. Osteoblasts were treated with or without Cytochalasin D and then loaded or unloaded by fluid shear stress at 12 dyne/cm². The nested realtime PCR and immunofluorescence were performed to detect the expression levels of COX-2 mRNA and protein in the osteoblasts，respectively. Statistical analyses were performed using twoway ANOVA.

Results

Cytoskeleton integrity disruption with Cytochalasin D had antagonistic effect on the expression of COX-2 gene induced by fluid shear stress （P ＜0.05）. Cytochalasin D had no significant effects on the expression levels of COX-2 mRNA and protein under unloaded condition （P ＞0.05）. Fluid shear stress significantly increased the expression levels of COX-2 mRNA and protein in osteoblasts （P ＜0.05）. Cytochalasin D significantly decreased the expression levels of COX-2 mRNA and protein in osteoblasts induced by fluid shear stress （P ＜0.05）.

Conclusion

Cytoskeleton integrity in osteoblasts is essential for the expression of COX-2 gene induced by fluid shear stress.

Key words: COX-2 gene, Fluid shear stress, Cytoskeleton, Osteoblasts

Chang-jing WU, You-rui LI, Ya-juan XU, Ling GUO, Yi-ping ZHANG, Qiang FU. Effect of cytoskeleton integrity on the expression of COX-2 gene in osteoblasts induced by fluid shear stress[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2009, 3(04): 403-409.

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URL: https://zhkqyxyjzz.cma-cmc.com.cn/EN/10.3877 / cma.j.issn.1674-1366.2009-04-009

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Figures/Tables 5

References 23

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组别	流体剪切力加载	细胞松弛素D
1	不加载	空白
2	加载1.5 h	空白
3	不加载	细胞松弛素D
4	加载1.5 h	细胞松弛素D

组别	流体剪切力加载	细胞松弛素D
1	不加载	空白
2	加载1.5 h	空白
3	不加载	细胞松弛素D
4	加载1.5 h	细胞松弛素D

引物	顺式序列（5'-3'）	反式序列（5'-3'）	引物大小
COX-2外引物	TGGAAAAGGTTCTTCTACGGAGAG	CAGGGATGTGAGGAGGGTAGATC	97 000
COX-2内引物	TAAGCGAGGACCTGGGTTCA	AAGGCGCAGTTTATGTTGTCTGT	309 000
GAPDH	CGTGTTCCTACCCCCAATGT	TGTCATCATACTTGGCAGGTTTCT	73 000

引物	顺式序列（5'-3'）	反式序列（5'-3'）	引物大小
COX-2外引物	TGGAAAAGGTTCTTCTACGGAGAG	CAGGGATGTGAGGAGGGTAGATC	97 000
COX-2内引物	TAAGCGAGGACCTGGGTTCA	AAGGCGCAGTTTATGTTGTCTGT	309 000
GAPDH	CGTGTTCCTACCCCCAATGT	TGTCATCATACTTGGCAGGTTTCT	73 000

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