SEARCH AND POSSIBLE USAGE OF FUNCTIONAL MOLECULES USING COLLAGEN-FORMING CULTURED CELLS

以大鼠肝脏来源的M细胞为实验材料。这些细胞有一个独特的特点，就是它们作为菌株保持不变。然而，在维生素C的存在下，它们在长期的培养中可以形成可见的胶原纤维。我们通过生物测定指导的方法分离了一种胶原网络形成的效应物。从冲绳采集的百日草叶提取物(日文名称：YYYYAMA-HAMAKUSAGI)的正丁醇可溶部分中分离出一种苯乙醇类化合物，Acteoside是一种有效的化合物，可以诱变M细胞形成的胶原纤维网络。该化合物使M细胞形成非常微小的网络，这可能有利于伤口修复，因为颗粒形成得很快。对苷元和酯类化合物的构效关系、邻苯二酚结构的研究似乎都是必需的。该化合物在植物中分布较均匀，在植物中含量普遍较高。这意味着Acteoside来自天然来源。另一方面，来自Alangium Plamfolium var叶提取物的乙酸乙酯可溶部分。Platanlfolium(日本名：Momijiounoki)，从福冈县采集到的一种黄酮类和苯甘油加合物被分离出来，作为一种有希望的化合物抑制胶原纤维的形成。黄酮类成分与木犀草素关系密切，另一部分为3-羟基-4-甲氧基苯甘油。完全抑制活性估计为2μg/ml。其他含儿茶酚功能基团的黄酮类化合物也表现出一定的抑制活性。从这些结果来看，该方法可用于寻找具有抑制或促进胶原纤维形成活性的功能化合物。

M cells derived rat liver were used for experiments. These cells have a unique feature that they are maintained as a strain. However, they form visible collagen fibers on prolonged cultivation under a presence of vitamin C. we conducted the isolation of an effector of collagen network formation by means of a bioassay-guided procedure.From n-BuOH-soluble fraction of the leave extract of Premna subscandens (Japanese name : Yaeyama-hamakusagi), collected in Okinawa, a phenylethanoid, acteoside was isolated as an effective compound which mutate the formation of collagen fiber networks formed by M cell. The compound made the M cell to form very minute networks which might be favorable on an occasion of remedy from wound due to rapid formation of granules. The investigation of structure and activity relationship, catechol moieties seemed to be required for both aglycone and ester portions. This compound is relatively well distributed in plant species and the contents in plants are generally high. This means that acteoside is available from natural sources.On the other hand, from the EtOAc-soluble fraction of leave extract of Alangium platamfolium var. platanlfolium (Japanese name : Momijiurinoki), collected in Fukuoka Pref., a flavonoid and phenylglycerol adduct was isolated as a promising compound to inhibit the formation of collagen fiber. The flavonoid portion was closely related to luteolin and the other part was 3-hydroxy-4-methoxyphenylglycerol. The activity was estimated to be at 2 μg/ml for complete inhibition. Other flavonoids with catechol-like functional group also exhibit some inhibitory activity.From these results, this method found to be used to search functional compounds to have the activity to inhibit or promote collagen fiber formation.