Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, Utah 84112-5820, USA.
Mol Pharm. 2011 Aug 1;8(4):1016-24. doi: 10.1021/mp100374n. Epub 2011 Jan 21.
Farnesyl pyrophosphate synthase (FPPS), an enzyme in the mevalonate pathway, is the inhibition target of alendronate, a potent FDA-approved nitrogen-containing bisphosphonate (N-BP) drug, at the molecular level. Alendronate not only inhibits osteoclasts but also has been reported to positively affect osteoblasts. This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures. Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate. Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability. FPPS siRNA is more potent than 10 μM alendronate, but less potent than 50 μM alendronate on reducing osteoclast viability. Despite FPPS knockdown, no significant changes were observed in osteoblast proliferation. FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition. However, compared with 50 μM alendronate dosing, FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation. Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization. Overall, results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways.
法呢基焦磷酸合酶(FPPS)是甲羟戊酸途径中的一种酶,是阿仑膦酸钠(一种强效的美国食品药品监督管理局批准的含氮双膦酸盐(N-BP)药物)在分子水平上的抑制靶点。阿仑膦酸钠不仅抑制破骨细胞,而且据报道还能积极影响成骨细胞。本研究评估了针对 FPPS 的 siRNA 与阿仑膦酸钠在破骨细胞和成骨细胞培养中的敲低效果。使用原代鼠骨髓细胞诱导的破骨细胞和前成骨细胞 MC3T3-E1 细胞系来评估抗 FPPS siRNA 与阿仑膦酸钠的作用。结果表明,在基于血清的培养中,FPPS mRNA 信息和蛋白敲低与破骨细胞活力降低相关。FPPS siRNA 比 10 μM 阿仑膦酸钠更有效,但比 50 μM 阿仑膦酸钠更有效,可降低破骨细胞活力。尽管 FPPS 被敲低,但成骨细胞增殖没有明显变化。FPPS 敲低显著促进成骨细胞分化,但不促进细胞矿化沉积。然而,与 50 μM 阿仑膦酸钠给药相比,FPPS siRNA 对成骨细胞没有细胞毒性作用,而对成骨细胞分化则产生显著作用。在测试浓度下,siRNA 和阿仑膦酸钠对培养的成骨细胞矿化均无显著影响。总的来说,结果表明,针对 FPPS 的 siRNA 可能有助于通过以不同的方式影响破骨细胞和成骨细胞,选择性地抑制破骨细胞介导的骨吸收并改善骨量维持。