protoporphyrin: E%cm=4900 coproporphyrin: uroporphyrin: E%cm=6667 E%cm=6517 Fig. 1. Changes of Absorption Spectra of Chloroform Extracts containing Pr
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2 protoporphyrin: E%cm=4900 coproporphyrin: uroporphyrin: E%cm=6667 E%cm=6517 Fig. 1. Changes of Absorption Spectra of Chloroform Extracts containing Protoporphyrin after Adding Zn. acet. and 12 Methanol Solution
3 Fig. 2. Changes of Fluorospectrophotometric Curves of Chloroform Extracts containing Protoporphyrin after Adding Zn. acet. and 12 Methanol Solution. Fig. 3. Changes of Fluorospectrophotometric Curves of Protoporphyrin Chloroform Solution after Adding Zn. acet. and 12 Methanol Solution
4 Table I. Changes observed in the Blood of Phlebotomized Anemic Rabbits. B. P. P. Bile Pigment Precursors E. S. Fe: Easily Split off Blood Iron Hb: Hemoglobin
5 赤 血 球 内Bile Fig. 4. Hematological Pigment and Chemical the Blood of Phiebotomized Table II. Precuraora Anemic Changes in Rabbits. Changes observed in the Blood of Rabbits following Administration of Phenylhydrazine HC1 (subcutaneous injection of phenylhydrazine HCl 50mg/kg) 1715
6 B. P. P.: Bile Pigment Precursors E. S. Fe: Easily Split off Blood Iron Hb: Hemoglobin * Concentration of NaCl Solution (%)
7 赤 血 球 内 Bile Pigment Precuraora 1717 Fig. 5. Hematological and Chemical Changes in the Blood of Rabbits following Administration of Phenylhydrazine HCl Table III. Changes observed in the stored Blood of Dogs. (stored at 4 Ž in A. C. D. solution)
8 1718 中 B. P. P.: E. S. Fe: Hb: * 川 昌 壮 Bile Pigment Precursors Easily Split off Blood Iron Hemoglobin Concentration of NaCl Solution (%) by Fennel's Method Fig.6. Hematological andchemical Changes in the stored Blood 12時 間 目の値 は1140μg/Hbで あ り.こ の 値 は保 存 前 の そ れ に 比 す れ ば 約80倍 に 当 る.赤 E. S. Feも6時 間 目を境 と し著増 した. 血 球内 of Dogs 赤 血 球 内E. P.の 変 動 は,保 存 開始 後2時 間 目よ りす で に か な り増 加 し,更 に12時 間 目まで増加 を続,けて前 値 の 約4倍 量 に 達 した 後 減少 した.
9 赤 血 球 内Bile Table IV. Changes observed at 37 Ž 7. Hematological of Dog after and the with in acid Chemical Incubation 100mg 37 Ž Precursors Blood with Changes at 1-Ascorbic the together (1-ascorbic Fig. Pigment in of 1719 Dog after 1-Ascorbic per 20ml the Blood the Incubation Acid of blood) 成 時 期 か らheme生 together globin生 Acid 成 乃 至hemo 成 を経 て 移 行 す る 過 程 を 否 定 出 来 な い.実 験 で はE. S. Fe の 増 加 は実 験 前値 へ の 恢 復 以 外 に 認 め 難 い た めE. らB. P. P.の 向を認 め難 P.の 増加か 増 加 の 間に増 加 傾 く,却 てB. 増 加 よ り お くれ てE. P. P.の S. Feの を み と め て い る.処 恢復 で血 色 素 よ り 胆 汁 色 素へ の 生成 過 程 に 生 ず る choleglobin, verdohemoglobin系 の 中 間 物 質 か ら のE. 赤血球 内遊 離coproporphyrinはE. P.の 増 加 の 著 しい時 に増 量 を 認 め た が, uroporphyrinは 認め られなかつ た. す る鉄 分 離 とbiliverdin抽 い こ と は,教 従 つ てB. 赤血 球 は保存6時 間 を 越 す と形 態 上 の 変 化が 著 し P. P.とE. S. Feの と か らporphyrinよ 消 長 が 相前 後す る こ りhemoglobin生 過 程 を 省 い て 直 ち にB. か6時 間 目に は認 め られな くな つ た. 断 定 す る 二 と は 危 険 で あ ろ う.教 E. S. Feの 先づ瀉 血貧 血家 兎 赤 血球 の場 合で は,網 状 赤 血球 の増加 に伴 いE. P.が 著 し く増 加 し,略 々消 長 を 等 し くす る が,そ E. S.Feと の 増 加 時 期 で はB. も最 も減少 し,次 い でE. 減少 に移 る時 期 に一 致 してB. P. P.及 び P.が 増 加 か ら P. P.の P. P.へ 成 を経ての の 過 程 への 移 行 を 室 の 田 淵11)は ら鉄 代 謝 の 面 か ら 新 生 赤 血 球 中 でheme生 括 並 び に 考 按 増 加 を認 め, 称 室 の 湯 浅 等 が 明 ら か に した 処 で あ り, く,算 定 が困難 とな り,又 網状 赤血 球 は 保 存前3% IV.総 S. Feと 出 の 難 易 は 平行的 で な 専 成 の 皮, 増 加 の 起 るこ とを証 明 して い る こ と もこ の 推 論 を 裹 付 け る も の で あ ろ う. 但 し, London, phyrinを I. M. et al.8)ら はC14-protopor 用 いporphyrinよ を 証 明 し,又Fischer, 乱tioporphyrin Iか りの 胆汁 色 素 への 分解 H. & Herrle, K. 19)ら は らphotochemical oxidationに E. S. Feも 増 加 す るが実 験 前 値 に 復す るに過 ぎず, よ り 胆 汁 色 素 の 生 成 を 証 明 して い る と こ ろ か ら 尚 そ その傾 向 もや やお くれ て い た.網 状 赤 血 球 とE. P. の 可 能 陸 も十 分 考 慮.よ る必 要 が あ ろ う. の消長 が略 々平 行 す る こ とは既 に 報 告16)さ れ て い る所 で あ る が,生 成 さ れ たE. P.か ら直 ちに 次 に 塩 酸phenylhydrazine処 に 於 て は 網 状 赤 血 球 とE. B. P. P.に 移行 す るか ど うか は本 実 験 か ら一 応 の 肯 行 し な い が,貧 定的所見 を推 論 す る こ とが 出来 るが, porphyrin生 共 に 増 加 す る.赤 血 に 伴 いB. 置 家兎 赤 血球 の 場 合 P.の 消長 は必 ず しも平 P. P.及 びE. S. Feは 血 球 中 の 血 色素 が 塩 酸phenylhy-
10
11 1) London, I. M., West, R., Shemin, D. & Rit tenberg, D.: J. Biol. Chem., 184: , ) Gray, C. H., Neuberger, A. & Sneath, P. H. A.. Biochem, J., 45: XVI-XVII, ) Gray, C. H. & Neuberger, A.: Biochem, J., 47: 87-92, ) London, I. M. & West, R.: J. Biol. Chem., 184: , ) London, I. M., West, R., Shemin, D. & Rit tenoerg, R.. J. Biol. Chem. 184: , ) Watson, C. J.: Arch. Inter. Med., 86: , ) Grinstein, M., Kamen, M. D., Wikoff, H. M. & Moore, C. V.. J. Biol. Chem., 182: , ) London, I. M., Yamasaki, M. & Gloria, S.: Fed. Proc., 10: 217, ) Granick, S. & Schrieck, H. G. V.. Proc. Soc. Exp. Biol. & Med., 88: , ) Grinstein, M.. J. Biol. Chem., 167: , ) Gardikas, C., Kench, J. E. & Wilkinson, J. F.: Biochem. J., 46: 85-88, ) Schwartz, S. & Wikoff, H. M.. J. Biol. Chem., 194: , ) Rimington, C. & Sveinsson, S. L.. Scand. J. Clin. Lab. Invest., 2: 209, ) Grinstein, M., Silva, J. A. & Wintrobe, M. M.: J. Clin. Invest., 27: , ) London, I. M.: J. Biol. Chem., 184: , ) Fischer, H. & Herrle, K.: Z. physiol. Chem., 251: 85-96, ) Lemberg, R. & Legge, J. W.. Hematin Co. mpounds and Bile Pigments, Interscience Pu blishers. INC., New York. P 598, 1949.
12 26) Fairly, N. H.: Brit. Med. J., 2: , Studies on Bile Pigment Precursors in Erythrocytes Part 2. Relationship between Bile Pigment Precursors and Free Porphyrin By Shoso NAKAGAWA in Erythrocytes From the First Depertment of Internal Medicine, Okayama University, Medical School (Director: Prof. K. Kosaka) In order to study the possibility of porphyrin to become bile pigment precursors, the relations between free porphyrin in erythrocytes, especially protoporphyrin, and intracorpu scular bile pigment precursors were exp3rimentally observed. The results were as follows: 1. A part of protoporphyrin which increased was observed to change quickly into bile pigment precursors in the erythrocytes of a phlebotomized anemic or phenylhydrazine HC1 administerei rabbits where reticulocytosis occurred. 2. This transformation of protoporphyrin into bile pigment precursors were also obser ved in preserving canine erythrocytes where reticulocytosis was caused by phlebotomies. 3. Judging from the variations of intracorpuscular easily split off blood iron, it seemed appropriate to consider that this transformation occurred after the formation of hems or hemoglobin. In the above exp3rlmml, however, of preserving erythrocytes of a phlebotomized anemic dag, the plainly direct transformation of protoporphyrin into bile pigment precursors were also observed. 4. In preserving canine erythrocytes with or without the addition of ascorbic acid, the increase of protoporphyrin and of bile pigment precursors were also observed. However, it is difficult to account for th3 relations betwaen these two; the increase of bile pigment pre cursors may be ascribab~e to the breakdown of hemoglobin. 5. Intracorpuscular free coproporphyrin was obsarvei to increase in those erythrocytes after phleiotom_??_es. phenylhydrazine HC1 aiministratioa, or preservation with the addition of ascorbic acid. This increase may more properly be ascribed to protoporphyrin in the forma tion procass of hemoglobin than to bile pigment precursors. On the other hand, free uropor phyrin in erythrocytes dii not show any significant variations. 6. Protoporphyrin mixed with chloroform extract did not hinder the microquantitative analysis of bile pigment precursors by fluorescence.
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