State Key Laboratory for Supramolecular Structure and Materials, Jilin University, Changchun 130012, China.
J Inorg Biochem. 2010 Apr;104(4):446-54. doi: 10.1016/j.jinorgbio.2009.12.018. Epub 2010 Jan 4.
The effects of sinomenine (SIN, an alkaloid extracted from the Chinese medicinal plant Sinomenium acutum used for centuries to treat rheumatic disease, including rheumatoid arthritis) on apatitic nucleation and matrix vesicle (MV)-induced mineral formation were compared with those of cysteine, levamisole, and theophylline. We found that SIN was not an inhibitor of tissue non-specific alkaline phosphatase (TNAP), a marker of biological mineralization, but confirmed that cysteine, levamisole, and theophylline were. Further, none of these four molecules directly affected the nucleation of hydroxyapatite (HA) formation, in contrast to pyrophosphate (PP(i)) which did. Incubation of 0.25-1.0mM cysteine, theophylline, or levamisole with MVs in synthetic cartilage lymph (SCL) containing AMP and Ca(2+), but not inorganic phosphate (P(i)), prolonged the induction time of mineral formation, apparently by inhibiting TNAP activity. SIN at the same levels neither inhibited TNAP activity nor affected the induction time of MV mineral formation. However, SIN did markedly delay MV-induced mineral formation in SCL containing P(i) (instead of AMP) in a manner similar to theophylline, but to a lesser extent than levamisole. Cysteine did not delay, in fact it slightly accelerated MV-induced mineral formation in Pi-containing SCL. These findings suggest that levamisole, SIN and theophylline may directly affect Ca(2+) and/or P(i) accretion during mineral formation; however, TNAP was not directly involved. The possible roles of annexins and other ion transporters, such as proteins of the solute carrier family implicated in Ca(2+) and P(i) influx are discussed.
将盐酸青藤碱（SIN，一种从中国药用植物青风藤中提取的生物碱，几个世纪以来一直用于治疗风湿性疾病，包括类风湿关节炎）对磷灰石成核和基质小泡（MV）诱导的矿化作用与半胱氨酸、左旋咪唑和茶碱的作用进行了比较。我们发现 SIN 不是组织非特异性碱性磷酸酶（TNAP）的抑制剂，TNAP 是生物矿化的标志物，但证实半胱氨酸、左旋咪唑和茶碱是 TNAP 的抑制剂。此外，这四种分子都没有直接影响羟基磷灰石（HA）形成的成核，而焦磷酸（PP（i））则有影响。将 0.25-1.0mM 半胱氨酸、茶碱或左旋咪唑与含有 AMP 和 Ca(2+)但不含无机磷（P(i)）的合成软骨淋巴液（SCL）中的 MV 孵育，延长了矿化形成的诱导时间，显然是通过抑制 TNAP 活性。相同浓度的 SIN 既不抑制 TNAP 活性，也不影响 MV 矿化的诱导时间。然而，SIN 在含有 P(i)（而不是 AMP）的 SCL 中明显延迟 MV 诱导的矿化形成，其方式类似于茶碱，但程度低于左旋咪唑。半胱氨酸不仅没有延迟，实际上它略微加速了 Pi 存在的 SCL 中 MV 诱导的矿化形成。这些发现表明，左旋咪唑、SIN 和茶碱可能直接影响矿化过程中 Ca(2+)和/或 P(i)的积累；然而，TNAP 并未直接参与。讨论了 annexin 及其他离子转运蛋白（如参与 Ca(2+)和 P(i)内流的溶质载体家族的蛋白质）的可能作用。
