Nutritional Value of S-Methylcysteine Sulfoxide (Methiin) (V) Investigations on effects of S-Methylcysteine Sulfoxide on Methionine-substitution and a

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1 Nutritional Value of S-Methylcysteine Sulfoxide (Methiin) (V) Investigations on effects of S-Methylcysteine Sulfoxide on Methionine-substitution and amino acid imbalance. Noriko Yagi, Machiko Takashima and Yoshinori Itokawa Department of Hygiene, Faculty of Medicine, Kyoto University, Kyoto Comparative studies on the effects of S-methylcysteine sulfoxide (SMCS), S-allylcysteine sulfoxide (SACS) and methionine on rats were carried out. Experiment No. 1 Young male Wistar rats were separated into five groups and five synthetic diets (amino acids mixture was used as protein source) were given. To group 1, methionine and cystine deficient diet; to group 2, SMCS supplemented to methionine and cystine deficient diet; to group 3, SACS supplemented to methionine and cystine deficient diet; to group 4, cystine supplemented to methionine deficient diet; and to group 5, complete amino acid mixture diet were given. In group 5, good growth was observed while body weight in group 1, 2, 3 and 4 showed progressive loss and after days these animals died. These facts suggested that SMCS, SACS and cystine had no methionine-substitution effect on the growth of methionine-cystine deficient rats. Experiment No. 2 Adult male Wistar rats were separated into six groups and six semisynthetic diets were given. Group 1, low protein diet (5% casein); group 2, high protein (15% casein); group 3, low protein plus 2% methionine; group 4, high protein plus 2% methionine; group 5, low protein plus 2% SMCS; group 6, high protein plus 2% SMCS containing diet. After two weeks of these dietary regimens, the animals were sacrificed and various examinations were carried out. Increase of liver triglyceride, abnormal pattern of fatty acids in liver and decrease in serum threonine were prominent in group 3. While in the other groups such abnormalities were not seen. From these findings, it was clarified that even when an excessive amount of SMCS was added to the low protein diet, this amino acid do not produce symptoms of amino acid imbalance as seen in rats fed excessive amount of methionine.

2 Table 1 Composition of Synthetic Diets (Experiment No. 1) Amino Acids Mixture* (in 1kg of diet) Basal amino acids mixture (sulfur containing amino acids free) To basal amino acids mixture followings were added; Group 1; Sucrose 6.93g, Group 2; S-Methylcysteine Sulfoxide 6.93g, Group 3; S-Allylcysteine Sulfoxide 6.93g, Group 4; Cystine 6.93g, Group 5; Methionine 6.93g. Besides amino acids mixture each diet contains the following; (in 1kg diet). Sucrose; 713g, Olive oil; 100.0g, Salt mixture; 40.0g**, Cellulose; 20.0g, Vitamin mixture; 5.0g**, Choline chloride; 2.0g. *Amino Acids composition was prepared according to Miyazaki17). **Salt mixture and Vitamin mixture were prescribed as described in a previous paper2). Table 2 Composition of Synthetic Diets (Experiment No. 2) (%) * Salt mixture and vitamin mixture were prepared as described in a previous paper2).

3 6 Rats Average Fig. 2 Growth Curves (Experiment No. 2) Group 1; 5% Protein Group 2; 15% Protein Group 3; 5% Protein+Methionine Group 4; 15% Protein+Methionine Group 5; 5% Protein+S-Methylcysteine sulfoxide Group 6; 15% Protein+S-Methylcysteine sulfoxide addition to basal amino acid mixture 5 Rats Average Fig. 1 Growth Curves. (Experiment No. 1) Table 3 Ratio of Liver, Kidney and Spleen Weight to Body Weight (Experiment No. 2) (%) the method of Student's "t" test.

4 596 日 は, 衛誌 15%タ (Jap. J. Hyg.) 第30巻 第6号 1976年2月 ン パ ク 食 群 ラ ッ ト (2, 4, 6群) に 比 較 し成 長 が 悪 か っ た Methiin 6群) を 添 加 し た ラ ッ ト(5, は, そ れ を 添加 しなか った ラ ッ ト (1, 2群) よ りも成 長 が お くれ る 傾 向 が み ら れ た 体 重 当 り の 肝 臓 腎臓 脾臓 重 量 は Table 3に 示 す 如 くで あ っ た 腎 臓 重 量 は Methiin thionine 5及 び6群 15%タ あ る い は Me- 添 加 食 を 摂 取 した3, で 増 加 し, 脾臓重量は ンパ ク に Methionine い は Methiin 4, あ る を 添 加 した飼 料 を 摂 取 した 群 で 増 加 が 認 め られ た 肉 眼 的 に は5%タ nine ンパ ク に Methio- 添 加 した 飼 料 を 摂 取 した ラ ッ ト (3群) の肝 臓 の色 調 が 白黄 色 に な っ て い た 2) 組織 学 的所 見 肝 臓 の 脂 肪 染 色 組 織 像 は, 3に Fig. 示 す と お りで あ っ た 5%タ Fig. 3 Microscopic picture of Livers. (Experiment No. 2) Sudan III stain. 100 A. Group 3. Fat accumulation is seen in periportal and B. Group 5. Fat accumulation is not seen in all hepatocytes. midzonal hepatocytes. Table Values 4 represent Significant the method Triglyceride mean±s.e. and Cholesterol difference of six (p<0.05) of Student's "t" rats. compared test. * SMCS: with Levels in Serum S-Methylcysteine group 2. Statistical and Liver (Experiment No. 2) sulfoxide. analysis was carried out according to

5 Table 5 Fatty Acids Composition in Livers (Experiment No. 2) (%) the method of Student's "t" test. Table 6 Serum Amino Acid Patterns (Experiment No. 2) the method of Student's "t" test. * MeSH; Methionine, ** SMCS; S-Methylcysteine Sulfoxide

6 1) M. Fujiwara, Y. Itokawa, H. Uchino and K. Inoue: Anti-hypercholesterolemic effect of a sulfur containing amino acid, S-Methyl-L-Cysteine sul-

7 foxide, isolated from cabbage. Experientia, 28, (1972). 2) Y. Itokawa, K. Inoue, S. Sasagawa and M. Fujiwara: Effect of S-Methylcysteine sulfoxide, S-Allylcysteine sulfoxide and related sulfur containing amino acids on lipid metabolism of experimental hypercholesterolemic rats. J. Nutrition, 103, (1973). (1969). 11) Wiebers, J. L. and Garner, H. R.: Use of S-Methylcysteine and cystathionine by Methionineless neurospora mutants. J. Bacteriology, 88, (1952). 15) Sperry, W. H. and Webb, M.: A revision of the 7) Stevens, C. M., Johnson, C. A. and Watanabe, R.: Preparation of S-Allyl-DL-Homocysteine and related compounds and tests of their grouth effects in rats. J. Biol. Chem., 212, (1955). 8) Weller, L. E., Luecke, R. W. and Sell, H. M.: Effect of derivatives of L-Cysteine on the grouth of Leuconstoc mesenteroides Biol. Med., 77, 3-5 (1951). P-60. Proc. Soc. Exp. 9) Smith, I. K. and Thompson, J. F.: Utilization of S-Methylcysteine and Methylmercaptan by Methionineless mutants of Neurospora and the Pathway of their conversion to Methionine. Biophys. Acta, 184, (1969). Biochim. 10) Moor, D. P., Thompson, J. F. and Smith, I. K.: Utilization of S-Methylcysteine and Methylmercaptan by Methionineless mutants of Neurospora and the pathway of their conversion to Methionine. Biochim. Biophys. Acta., 184, (1964). 14) Abell, L. L., Levy, B. B., Brodie, B. B. and Kendall, F. E.: A simplified method for the estimation of total cholesterol in serum and demonstration of its specificity. J. Biol. Chem., 195, Schoenheimer Sperry methods for cholesterol determinations. J. Biol. Chem., 187, (1950). 16) Sardesai, W. M. and Manning, J. A.: The determination of triglycerides Clin. Chem., 14, (1968). in plasma and tissues.

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