| Research Abstract |
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
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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
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