Graduate School of Biotechnology, Kyung Hee University, Yongin, Korea.
J Med Food. 2012 Apr;15(4):384-90. doi: 10.1089/jmf.2011.1827. Epub 2012 Feb 8.
The prophylactic effects of Hijikia fusiforme on bone metabolism were examined using in vitro indices of bone formation and resorption. As the indices of bone formation, osteoblast proliferation and differentiation were measured through mitochondrial enzyme activity [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay] and bone marker alkaline phosphatase (ALP) activity. The aqueous extract of H. fusiforme stimulated the proliferation of the human osteoblast-like cell line MG63 and the ALP activity of the mouse osteoblast-like cell line MC3T3-E1. Moreover, extracellular matrix mineralization was accelerated by the addition of H. fusiforme. As the indices of bone resorption, differentiation of the murine macrophage/osteoclast precursor cell line RAW 264.7 by receptor activator of nuclear factor-κB ligand (RANKL) was measured as tartrate-resistant acid phosphatase-positive multinucleated cells, which were suppressed by H. fusiforme. Additionally, H. fusiforme lowered the secreted amount of RANKL that is required for the osteoclastic differentiation and activation, but the amount of osteoprotegerin as a decoy receptor for RANKL was not affected. The bone-protective effects of H. fusiforme in estrogen-deficient ovariectomized rats were also investigated. Osteoporosis was induced in female Sprague-Dawley rats by ovariectomy for 15 weeks, and then H. fusiforme was orally administered at a dose of 100 mg/kg of body weight every day for 6 weeks. Bone mineral density in the group orally administered H. fusiforme was increased, compared with ovariectomized rats, but not significantly (P>.05). Oral administration of H. fusiforme improved microarchitecture of bone in terms of bone volume (bone volume/total volume ratio) and trabecular separation (P<.05). The amounts of osteocalcin and C-telopeptide type I collagen in serum were measured as the biomarkers for bone formation and resorption. The level of osteocalcin in serum was increased, but not significantly. However, the level of C-telopeptide type I collagen in serum was significantly decreased (P<.05). When the results are taken together, the present study indicates that H. fusiforme might be useful in the treatment of osteoporosis.
本研究采用体外骨形成和骨吸收指标来检测海蕴对骨代谢的预防作用。作为骨形成的指标，通过线粒体酶活性[3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐测定法]和骨标志物碱性磷酸酶（ALP）活性来测量成骨细胞增殖和分化。海蕴的水提物可刺激人成骨样细胞系 MG63 的增殖和小鼠成骨样细胞系 MC3T3-E1 的 ALP 活性。此外，海蕴可加速细胞外基质矿化。作为骨吸收的指标，通过核因子-κB 受体激活剂配体（RANKL）分化鼠巨噬细胞/破骨细胞前体细胞系 RAW 264.7 作为抗酒石酸酸性磷酸酶阳性多核细胞来测量，海蕴可抑制该细胞分化。此外，海蕴降低了破骨细胞分化和激活所需的 RANKL 的分泌量，但不影响 RANKL 的诱饵受体骨保护素的量。还研究了海蕴对去势雌性大鼠的骨保护作用。通过去势 15 周诱导雌性 Sprague-Dawley 大鼠骨质疏松症，然后每天口服 100mg/kg 海蕴 6 周。与去势大鼠相比，口服海蕴的大鼠骨密度增加，但差异无统计学意义（P>.05）。口服海蕴可改善骨的微观结构，包括骨量（骨量/总体积比）和小梁分离（P<.05）。作为骨形成和骨吸收的生物标志物，测量了血清中骨钙素和 I 型胶原 C 端肽的量。血清骨钙素水平升高，但无统计学意义。然而，血清 I 型胶原 C 端肽水平显著降低（P<.05）。综上所述，本研究表明海蕴可能有助于治疗骨质疏松症。
