enlarged in scurvy, but McCarrison [1919], working on guinea-pigs dying of this disease, found that there was at the same time an absolute

Transcription

1 478 6I2.45I:6I6*392/3 THE ADRENALINE CONTENT OF THE SUPRARENAL GLANDS IN SCURVY AND IN INANITION. BY WALTER DEUTSCH AND WALTER SCHLAPP. (From the Department of Physiology, University of Manchester.) (Received December 29, 1934.) THE diminution in the amount of ascorbic acid which has frequently been described in the suprarenal gland in scurvy has directed attention to the adrenaline content in the same condition, the occurrence of both substances in the same gland suggesting a relationship between them. Thus, it is conceivable that ascorbic acid might be necessary for stabilization on account of its protective action when in a reduced state, with respect to the oxidation of adrenaline [Szent-Gy6rgyi, 1928; Heard and Raper, 1933]. v. Euler and Klussmann [1933] suggest, less definitely, that there may be a close physiological relationship between the two substances. It is well known that the suprarenal glands are usually found to be enlarged in scurvy, but McCarrison [1919], working on guinea-pigs dying of this disease, found that there was at the same time an absolute reduction in the amount of adrenaline present. Mouriquand and Leulier [1927], working with animals with a less severe degree of the condition, were unable to confirm McCarrison's observation, though they described a diminution in the amount of adrenaline per gram of gland substance. This latter observation has been repeated in experiments described by v. Euler and Klussmann [1933]. In our experiments young guinea-pigs weighing between 250 and 500 g. were kept in separate cages in the laboratory on a standard diet made up of three parts of bran, twenty-seven parts of rolled oats and one and a half parts of dried egg yolk, to which was added 0 5 ml. of pure cod-liver oil per animal per day. There was, in addition, a daily allowance of 25 g. of fresh lettuce leaves for each animal, and abundant fresh water

2 479 was provided. The experimental animals were deprived of green food after a preliminary period of observation. In from 14 to 21 days, when they showed by loss of weight and by the other characteristic signs that scurvy had developed, the animals together with the normal controls were killed by a blow on the head, and the suprarenal glands were dissected out. After the removal of as much of the adherent tissue as possible the pairs of glands were weighed, ground up in a mortar with : C) P Fig. 1. clean sand, and extracts were prepared according to the method described by Folin, Cannon and Denis [1913]. The adrenaline content of these extracts was estimated on the blood-pressure of decapitated cats. Solutions of adrenaline hydrochloride (Burroughs Wellcome "Soloid" brand) made up with 09 p.c. sodium chloride and the same quantities of hydrochloric acid and of sodium acetate as were contained in the gland extracts, were used as standards of comparison. The adrenaline content was expressed as y of adrenaline hydrochloride per gram of whole gland. The adrenaline content of the normal suprarenal gland was found to be from 250 to 320y adrenaline hydrochloride per gram. The figures for PH. LXXXIII. 31

3 480 W. DEUTSCH AND W. SCHLAPP. six animals are graphically represented in Fig. 1. The values thus obtained are in sufficiently good agreement with 270y of Leulier and Gojon [1927] and 230y of McCarrison [1919]. In a series of ten animals in which scurvy of varying degrees of severity had been induced, uniformly lower values were obtained, as may be seen in Fig. 1. In the majority of these animals the weights of the glands were found to be greater than would be expected in normal animals of the same initial weight (Fig. 2). The increase is, however, in general -~~ ~~~~~~~~~~~~~~~~~~~~ 'l30 2 0lll 03 NORMAL SCURVY 4 'Zm '200 Fig. 2. less than would account for the decreased adrenaline content per gram, so that there is an abso lute diminution as has been previously shown by McCarrison [1919]. ADRENALINE IN INANITION. But a comparison of the growth curves of the scorbutic animals with those of the controls (Fig. 3), and a study of the general demeanour of the former, soon raises doubts as to whether this diminution can be regarded as characteristic of the scorbutic condition alone. For it becomes clear at once that the scorbutic animals are suffering to a considerable extent from inanition, while the controls are growing normally. Indeed, it can be seen from Fig. 1 that there is a close relationship between the percentage loss of weight and the adrenaline content of the suprarenal gland. Though loss of weight is of course an indication of the severity of the scurvy, it was obviously of interest to study the effect of inanition,

4 produced by a reduction of the food supply, on the adrenaline content of the glands. For this purpose the average daily food consumption of six guinea-pigs was determined. While the full allowance of green food was maintained, the total food was reduced by an amount not exceeding 50 p.c. for from 6 to 8 days, while adequate supplies of drinking water were provided. In this way it was possible to reproduce in the animals growth curves which closely resembled those seen in the, less seriously b5o 0,Z 400 Normal Inanition 350 Scurvy - I =M-.LM I I I I days Fig. 3. affected scorbutic ones (Fig. 3). In spite of this they were in a better state of general health than the corresponding ones with scurvy. The results of adrenaline estimations in these partially starved animals are represented in Fig. 1. There is in every instance a reduction. While it has not been possible to examine animals which have become moribund from lack of food it is evident that an important factor in the production of the reduced adrenaline content in scurvy must be the associated inanition. It is by no means without the bounds of possibility that this is the only factor. 31-2

5 482 W. DEUTSCH AND W. SCHLAPP. AscORBIC ACID IN INANITION. In spite of the fact that the animals suffering from inanition were receiving a full allowance of green food, it seemed worth while to make estimations of the ascorbic acid content of their glands. The method used was that of microtitration of a solution of 2-6-dichlorophenol-indophenol with trichloroacetic acid extracts of the glands as has been described by Birch, Harris and Ray [1933]. The solution of the dye was standardized against pure ascorbic acid. In a series of eight partially starved animals the values obtained ranged from 1*8 to 2 4 mg. of ascorbic acid per gram of gland, and there was no significant difference between these figures and those obtained from normal controls. The amount of ascorbic acid present in the glands of animals which have been on a scorbutic diet for 14 days is given by Harris and Ray [1933] as 021 mg. per gram of gland. These experiments indicate therefore that a reduction in the adrenaline content can take place without there being a reduction in ascorbic acid. They do not support the view that there is a close physiological relationship between the two substances. SUMMARY. 1. There is an absolute reduction in the adrenaline content of the suprarenal glands of guinea-pigs in scurvy. 2. There is a reduction in the adrenaline content of the suprarenal glands of guinea-pigs in inanition, and this must be an important factor in the reduction found in scorbutic animals. 3. There is no significant change in the ascorbic acid content of the suprarenalglandsininanition when sufficient greenfoodis given in the diet. 4. There does not appear to be any close physiological relationship between ascorbic acid and adrenaline. These experiments were undertaken at the suggestion of Prof. H. S. Raper, to whom we wish to express our indebtedness for his continued interest and advice. The work has been made possible for one ofus (W. D.) by grants from the Academic Assistance Council, and from the Rockefeller Foundation. REFERENCES. Birch, T. W., Harris, L. J. and Ray, S. N. (1933). Biochem. J. 27, 590. v. Euler, H. and Klussmann, E. (1933). Z. phyaiol. Chem. 217, 167. Folin, 0., Cannon, W. B. and Denis, W. (1913). J. biol. Chem. 13, 477. Harris, L. J. and Ray, S. N. (1933). Biochem. J. 27, 580. Heard, R. D. and Raper, H. S. (1933). Ibid. 27, 360. Leulier, A. and Gojon, P. (1927). C.R. Soc. Biol., Paris, 96, 547. McCarrison, R. (1919). Brit. Med. J. 2, 200. Mouriquand, G. and Leulier, A. (1927). C.B. Soc. Biol., Paris, 98, 548. Szent-Gyorgyi, A. (1928). Biochem. J. 22, 1387.