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Fasudil hydrochloride induces osteoblastic differentiation of stromal cell lines, C3H10T1/2 and ST2, via bone
Ippei Kanazawa1, Toru Yamaguchi, Shozo Yano
1Department of Internal Medicine 1, Shimane University Faculty of Medicine, Izumo, Japan. ippei.k@med.shimane-u.ac.jp
Abstract:
Rho-kinase (ROK), downstream of the mevalonate pathway, is detrimental to vessels, and suppressing its activity is a target for the treatment of human disease such as coronary artery disease and pulmonary hypertension. Recent studies have shown that ROK has a crucial role in bone metabolism. However, the role of ROK in stromal cells is still unclear. The present study was undertaken to investigate the effect of a ROK inhibitor, fasudil hydrochloride, on stromal cell lines, C3H10T1/2 and ST2. In both cells, Fasudil significantly stimulated alkaline phosphatase activity and enhanced cell mineralization. Moreover, fasudil significantly increased the mRNA expression of collagen-I, osteocalcin, and bone morphogenetic protein-2 (BMP-2). Supplementation of noggin, a BMP-2 antagonist, significantly reversed the fasudil-induced collagen-I and osteocalcin mRNA expression in both cells. These findings suggest that fasudil induces the osteoblastic differentiation of stromal cells via enhancing BMP-2 expression, and that this drug might be beneficial for not only atherosclerosis but also osteoporosis by promoting bone formation.
Insights
Fasudil hydrochloride, a Rho-kinase (ROK) inhibitor, promotes osteoblastic differentiation in stromal cells by increasing bone morphogenetic protein-2 (BMP-2) expression. This suggests potential therapeutic benefits for osteoporosis and atherosclerosis.
Area of Science:
- Biochemistry
- Cell Biology
- Bone Metabolism
Background:
- Rho-kinase (ROK) is implicated in vascular health and bone metabolism.
- The specific role of ROK in stromal cells remains largely undefined.
- ROK inhibition is a therapeutic target for cardiovascular diseases.
Purpose of the Study:
- To investigate the effect of the ROK inhibitor fasudil hydrochloride on osteoblastic differentiation in C3H10T1/2 and ST2 stromal cell lines.
- To elucidate the role of BMP-2 signaling in fasudil-induced stromal cell differentiation.
Main Methods:
- Treatment of C3H10T1/2 and ST2 cells with fasudil hydrochloride.
- Assay of alkaline phosphatase activity and cell mineralization.
- Quantitative real-time PCR analysis of collagen-I, osteocalcin, and BMP-2 mRNA expression.
- Treatment with noggin, a BMP-2 antagonist, to assess pathway involvement.
Main Results:
- Fasudil significantly increased alkaline phosphatase activity and cell mineralization in both cell lines.
- Fasudil treatment upregulated mRNA expression of collagen-I, osteocalcin, and BMP-2.
- Noggin supplementation reversed the fasudil-induced increase in collagen-I and osteocalcin mRNA levels, indicating BMP-2 dependency.
Conclusions:
- Fasudil hydrochloride induces osteoblastic differentiation of stromal cells.
- This differentiation is mediated through the enhancement of BMP-2 expression.
- Fasudil may hold therapeutic potential for osteoporosis and atherosclerosis by promoting bone formation.
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