Basic and clinical research for periodontal disease of physiologically vasoactive substances extracted from skeletal muscle of fur seal

我们之前研究了从海狗骨骼肌水解物中提取的肽样生理活性物质的外周血管扩张作用，发现肽样物质显示犬股动脉和腹部皮肤的血流量增加（Matsukawa等人，Bull.）。日本。Soc。科学。鱼类。40:1139,1974)。此外，这种肽样物质会增加犬的牙龈血流量（Tsujitani, Kanagawa Shigaku 18: 125, 1983），这表明这种特性被认为对发炎的牙周组织有益。当制备含有肽样物质的牙膏并在临床实践中将其应用于牙周病时，观察到牙龈状况的显着改善。随着时间的推移，红肿、引流、出血和牙袋深度的临床改善明显（Hiyama et J. Periodontol. 53: 639,1982）。然而，从海狗骨骼肌中提取的更多类骨物质的确切结构和/或成分尚不清楚，其结构与生理活动的关系尚不清楚。为了进一步了解该物质的药理作用，本研究重点建立从海狗骨骼肌中提取该物质的方法。测试了以下提取步骤。第一步是确定从海狗肌肉中提取该物质的效率；二是从粗提物中分离活性物质和提取分馏物的方法。血管活性物质在小于1000mw的分子范围内大量存在，而在大于1000mw的分子范围内则不存在。在分离方法方面，采用Amberlite XAD-2树脂吸收层析法，可以在不损失血管活性的情况下从含有高分子和低活性组分的粗提液混合物中分离出高活性组分。从海狗骨骼肌中提取的粗混合物中，经安贝岩XAD-2树脂层析得到的组分1，按照电密度离子交换层析进一步大致分为A ~ D 4个组分。对各组分进行表征，得到如下结果：部分A、B、C、D分别被鉴定为5′-ADP、5′-AMP、肌肽和鹅胺或组胺。据推测，从海狗骨骼肌中提取的肽样物质（粗混合物）对周围循环疾病的改善是由于所鉴定的血管活性物质，特别是腺苷衍生物之间的相互作用。少

We have previously examined peripheral vasodilating action of peptide-like physioactive substance extracted from the hydrolysate of skeletal muscle of fur seal, and found that the peptide-like substance shows an increase in blood flow of canine femoral artery and of abdominal skin (Matsukawa et al., Bull. Jpn. Soc. Sci. Fish. 40: 1139,1974). Furthermore, the peptide-like substance produces an increase in canine gingival blood flow (Tsujitani, Kanagawa Shigaku 18: 125, 1983), suggesting that this property is postulated to be beneficial to inflamed periodontal tissue. When a dentifrice containing the peptide-like substance was prepared and applied to periodontal disease in clinical practice to test this, significant improvement in the gingival conditions was observed. As time progressed, distinct clinical improvement in redness, swelling, drainage, bleeding and pocket depth was noted (Hiyama et al., J. Periodontol. 53: 639, 1982). However, precise structure and/or constituent of the pept … More ide-like substance extrated from skeletal muscle of fur seal are unknown, and relation between the structure and physiological activity requires clarification. To gain further insight into the pharmacological effect of the substance, the present study focuses on establishment of the method for extraction of the substance from skeletal muscle of fur seal.The following extractive steps were tested. The first step was to determine the procedure for the extractive efficiency of the substance from the muscles of fur seals; and the second was to detect the method of fractionating the active substance and of collecting the fractionate substance from crude extract. The vasoactive substances exsisted abundantly in the fraction of the molecular range less than 1,000 M.W., but not in that more than 1,000M.W. As for fractionating method, utilization of Amberlite XAD-2 resin absorptive chromatography was the best way to separate the high active fraction without a loss of vasoactive activity from crude extract mixture containing high molecular- and low active-fraction.The fraction 1 fractionated by amberlite XAD-2 resin chromatography from crude mixture extracted from skeletal muscle of fur seals was further divided broadly into 4 fractions (A to D) in compliance with electrical density ion exchange chromatography. Each fraction was characterized and following results were obtained. Fraction A, B, C, or D was identified as 5'-ADP,5'-AMP, carnosin and anserine, or histamine, respectively. it is postulated that improvement of peripheral circulatory disorders by the peptide-like substance (crude mixture) extracted from skletal muscle of fur seals is due to the interaction among the vasoactive substances, particularly adenosine derivatives, identified. Less