Suzuki Nobuo, Hayakawa Kazuichi, Kameda Takayuki, Triba Akira, Tang Ning, Tabata Makoto J, Takada Koji, Wada Shigehito, Omori Katsunori, Srivastav Ajai K, Mishima Hiroyuki, Hattori Atsuhiko
Noto Marine Laboratory, Institute of Nature and Environmental Technology, Kanazawa University, Noto-cho, Ishikawa 927-0553, Japan.
Life Sci. 2009 Mar 27;84(13-14):482-8. doi: 10.1016/j.lfs.2009.01.008. Epub 2009 Feb 3.
We previously demonstrated that monohydroxylated polycyclic aromatic hydrocarbons (OHPAHs) bound to a human estrogen receptor (ER) by a yeast two-hybrid assay, but polycyclic aromatic hydrocarbons did not have a binding activity. Therefore, the direct effect of 3-hydroxybenz[a]anthracene (3-OHBaA) and 4-hydroxybenz[a]anthracene (4-OHBaA) on osteoclasts and osteoblasts in teleosts was examined. As a negative control, 1-hydroxypyrene (1-OHPy), which has no binding activity to human ER, was used.
The effect of OHPAHs on osteoclasts and osteoblasts was examined by an assay system using teleost scale as each marker: tartrate-resistant acid phosphatase for osteoclasts and alkaline phosphatase for osteoblasts. Changes in cathepsin K (an osteoclastic marker) and insulin-like growth factor-I (IGF-I) (an osteoblastic marker) mRNA expressions in 4-OHBaA-treated goldfish scales were examined by using a reverse transcription-polymerase chain reaction.
In both goldfish (a freshwater teleost) and wrasse (a marine teleost), the osteoclastic activity in the scales was significantly suppressed by 3-OHBaA and 4-OHBaA, although 1-OHPy did not affect the osteoclastic activity. In reference to osteoblasts, the osteoblastic activity decreased with both 3-OHBaA and 4-OHBaA and did not change with the 1-OHPy treatment. However, 17beta-estradiol (E(2)) significantly increased both the osteoclastic and osteoblastic activities in the scales of both goldfish and wrasse. The mRNA expressions of both cathepsin K and IGF-I decreased in the 4-OHBaA-treated scales but increased in the E(2)-treated scales.
The current data are the first to demonstrate that 3-OHBaA and 4-OHBaA inhibited both osteoclasts and osteoblasts and disrupted the bone metabolism in teleosts.
我们之前通过酵母双杂交试验证明了单羟基化多环芳烃(OHPAHs)与人雌激素受体(ER)结合,但多环芳烃没有结合活性。因此,研究了3 - 羟基苯并[a]蒽(3 - OHBaA)和4 - 羟基苯并[a]蒽(4 - OHBaA)对硬骨鱼破骨细胞和成骨细胞的直接影响。作为阴性对照,使用了对人ER无结合活性的1 - 羟基芘(1 - OHPy)。
通过使用硬骨鱼鳞片作为每个标志物的检测系统来研究OHPAHs对破骨细胞和成骨细胞的影响:破骨细胞用抗酒石酸酸性磷酸酶作为标志物,成骨细胞用碱性磷酸酶作为标志物。通过逆转录 - 聚合酶链反应检测4 - OHBaA处理的金鱼鳞片中组织蛋白酶K(一种破骨细胞标志物)和胰岛素样生长因子 - I(IGF - I)(一种成骨细胞标志物)mRNA表达的变化。
在金鱼(一种淡水硬骨鱼)和隆头鱼(一种海水硬骨鱼)中,3 - OHBaA和4 - OHBaA均显著抑制鳞片中的破骨细胞活性,而1 - OHPy对破骨细胞活性没有影响。关于成骨细胞,3 - OHBaA和4 - OHBaA均使成骨细胞活性降低,而1 - OHPy处理则没有变化。然而,17β - 雌二醇(E₂)显著增加了金鱼和隆头鱼鳞片中的破骨细胞和成骨细胞活性。在4 - OHBaA处理的鳞片中,组织蛋白酶K和IGF - I的mRNA表达均下降,但在E₂处理的鳞片中则增加。
目前的数据首次证明3 - OHBaA和4 - OHBaA抑制硬骨鱼的破骨细胞和成骨细胞,并扰乱其骨代谢。