[Effect of Solanum melongena on experimental atheromatosis. VI. Enzyme histochemical, physiopathological and chemical studies on cholesterol-induced atheromatosis in rabbits. Conclusions (author's transl)].

QUESTION

The mode of action of Solanum Melongena (Sol. Mel.) in its inhibitory effect on the development of atheromatous plaques in cholesterol-induced atheromatosis in rabbits is still unexplained. In this paper the author reports his enzyme histochemical, physiopathological and chemical observations drawing conclusions for future experiments.

MATERIALS AND METHODS

Animals and experimental conditions were the same as described in the recent studies (see literature no. 2, 15, 16, 17, 18). 1. Enzyme histochemistry: The aortas were removed immediately after death, placed in Holt's solution, freed from the advential fatty tissue, cut into 5-6 mm high cylinders and either processed instantly or preserved in the same fluid at +2 degrees C. From the individual portions cryostat sections (10 mum thick) were cut after the method of Pearse. They were mounted to cover glasses (22 X22) and placed in substrate-filled specially produced cuvettes. The capacity of such a cuvette was 10 ml with room for 4 cover glasses each 4 mm apart. For protection against vaporization small caps of plastic foil were used. The method given by GLOCKNER and SCHMIDT (7) who recently reported their studies in the rat aorta, yielded satisfactory results in small arteries. In thereabbit aorta, however, the results varied markedly, which may be due to greater thickness of the vascular wall. Consequently, despite of all its advantages, this technique could not be employed in this study. The following enzymes were demonstrated: hydrolases: phosphomonoesterase I--3.1.3.1. with naphthol AS-sulphate after BURSTONE in the modification of LOJDA phosphomonoesterase II--3.1.3.2. after BURSTONE as modified by LOJDA with naphthol AS-phosphate and hexazonium pararosanilin leucine aminopeptidase--3.4.1.1. technique according to NACHLAS et al. with leucine beta-naphthylamide lipase--3.1.1.3. after GOMORI (Tween 80) adenosine-5-phosphatase (5'nucleotidase)--3.1.3.5. after GOMORI succinic dehydrogenase--1.3.99.1.--method according to NACHLAS et al. NAD (NADH and NADP (NADPH) diaphorase--1.4.1.2.--1.4.1.4., technique of SCARPELLI, HESS and PEARSE as modified by LOJDA (see article) Substratefree or heatinactivated controls were prepared for each histochemical demonstration. Several additional series with phenazine methosulphate (PMS) as an intermediate electron acceptor or such without PMS as well as preparations with KCN or without KCN were examined for the demonstration of anaerobic dehydrogenases. However, to avoid the nothing dehydrogenase effect only PMS-processed specimens were evaluated. For histological information several additional sections of the tissue samples were stained with haematoxylin-eosin or with elastica-van Gieson and Weigert's iron haematoxylin in combination. The enzyme histochemical preparations were embedded in Kaiser's glycerine gelatin, the histological sections were embedded in Caedox. The test animals were grouped like in the recent experiments (see literature no. 2, 15, 16, 17, 18). 2...