Effect of cholesterol administration on the state of the adrenal cortex in severe stress

The two weeks' effect of cholesterol on the rat adrenal cortex response under conditions of a severe stress was studied. In comparison with control, the degree of the gland activation and its alteration was found to decrease. It is suggested that the effect of cholesterol depends upon the action on the hypothalamus, and upon the adrenocortical tissue as well. The action of cholesterol on the adrenocortical tissue is bound with the cholesterol capacity to retard peroxydation reactions.     

Change in the adrenal cortex of rats in stress after hypophysectomy, thyroidectomy and castration

To elucidate factors producing destructive changes in the adrenal cortex under severe stress, the following experiments were performed: 1) severe combined stress (KC) including unilateral adrenalectomy and fixation in supin position for 24 h (control group); 2)hypophysectomy KC; 3) thyroidectomy KC; 4) bilateral orchiectomy KC; 5) injection of vitamine "E" for 14 days + KC. The adrenals were studied by a complex of morphometrical and histochemical methods. Hypophysectomy prevented estructive changes in the adrenal cortex under KC. This effect was not connected with hypofunction of the thyroid gland and the testes, as their excision (in rats without hypophysectomy) did not prevent the development of necrobiotic changes in the adrenal cortex. A suggestion is made on a "corticolytic" effect of ACTH which reveals itself under severe stress. Destruction of adrenocorticocytes is supposed to result from activation of peroxide oxidation of lipids. This is indirectly supported by a large number of lipofuscin containing cells, by decrease in the activity of alkaline phosphatase and ATPh-ase, by activation of acid phosphatase and also by decreasing alterative phenomena at the injection of vitamine "E".     

Substrate and inhibitor specificity of the cholesterol oxidase in bovine adrenal cortex

1. Cholesteryl 3β-sulphate is oxidized in vitro by preparations of bovine adrenal-cortex mitochondria to pregnenolone sulphate and isocaproic acid (4-methyl-pentanoic acid) without hydrolysis of the ester linkage. 2. Free cholesterol is the preferred substrate for adrenal-cortex cholesterol oxidase; the apparent K_m for cholesteryl sulphate is 500μm and for free cholesterol 50μm under the same conditions. 3. Cholesteryl 3β-acetate is hydrolysed by bovine adrenal-cortex mitochondria in vitro to free cholesterol, which is subsequently oxidized to more polar steroids and isocaproic acid. Evidence was obtained that other cholesterol esters behave similarly. Cholesterol esters may thus act as precursors of steroid hormones. 4. Cholest-4-en-3-one is only poorly oxidized to isocaproic acid and more polar steroids and thus is probably not a significant precursor of steroid hormones. 5. Cholesteryl esters inhibit the oxidation of cholesterol competitively (K_i for cholesteryl phosphate 28μm, for cholesteryl sulphate 110μm, for cholesteryl acetate 65μm) but pregnenolone esters do not inhibit this system. 6. Pregnenolone and 20α-hydroxycholesterol (both metabolites of cholesterol in this system) inhibit the oxidation of cholesterol non-competitively. K_i for pregnenolone is 130μm and K_i for 20α-hydroxycholesterol is 17μm. 7. 25-Oxo-27-norcholesterol inhibits cholesterol oxidation non-competitively (K_i16μm). A number of other Δ⁵-3β-hydroxy steroids inhibit cholesterol oxidation and evidence was obtained that the 3β-hydroxyl group was necessary for inhibitory activity. 8. Pregnenolone, 20α-hydroxycholesterol and 25-oxo-27-norcholesterol inhibit oxidation of cholesteryl sulphate by this system but their sulphates do not. 9. 3β-Hydroxychol-5-enoic acid, 3α-hydroxy-5β-cholanic acid and 3β-hydroxy-22,23-bisnorchol-5-enoic acid stimulated formation of isocaproic acid from cholesterol. 10. No evidence was obtained that phosphorylation or sulphation are obligatory steps in cholesterol oxidation by adrenal-cortex mitochondria. 11. The cholesteryl 3β-sulphate sulphatase of bovine adrenal cortex was found mostly in the microsomal fraction and was inhibited by inorganic phosphate.     

Histochemical study of the adrenal cortex of the rat after epiphysectomy]

The authors investigate the effects of pinealectomy on male rat's adrenal cortex. Fat material accumulates in the fascicular and reticular zones; different enzyme activities show light variations in operated animals. The frequent presence of mitochondrial multilamellar structures in pinealectomized rats is discussed.     

Lipid peroxidation in the adrenal cortex during exhausting stress

Under prolonged stress which is connected with exhaustion of functional resources of adrenal cortex the activation of lipid peroxidation processes in this gland was found. It is possible that the reason for such lipid peroxidation activation is the decrease in the content of adrenal cortex ascorbic acid and alpha-tocopherol.     

Specificity of the cholesterol side-chain cleavage enzyme system of adrenal cortex

Incubation of lanosta-8, 24-dien-3β-o1-1,2- ³H and lanost-8-en-3β-o1-1, 2-³H with an adrenocortical bovine mitochondrial acetone-dried preparation did not yield any significant ( < 0.01%) 3β-hydroxy-4, 4, 14-trimethyl-5α-pregn-8-en-20-one. Under the same conditions cholesterol-1,2-³H yielded 8.3% pregnenolone. Incubation of (20S?) — 17α, 20-dihydroxycholesterol-7-³ H yielded 0.6 to 1.6% (20SS?, 22R?) — 17α, 20, 22-trihydroxycholesterol, 1.0 to 3.2% of 17α-hydroxypregnenolone, but no significant ( < 0.02%) (20S, 22S)-17α, 20, 22-trihydroxycholesterol. In another experiment incubation of cholesterol-1, 2-³H yielded 5% pregnenolone, 0.5% 17α-hydroxypregnenolone, 0.2% (20R?,22R?)-20, 22-dihydroxy-cholesterol, but no significant ( < 0.01%) 17α-hydroxy-cholesterol, (20S?) -17α, 20-dihydroxycholesterol or (20S?, 22R?)-17α, 20, 22-trihydroxycholesterol.     

Autoradiographic study of the incorporation of tritiated cholesterol into the zona reticularis of the rat adrenal cortex

Summary In the zona reticularis of the rat adrenal cortex three types of cells are to be observed: light, dark, and very dark cells. A time study of the incorporation of cholesterol-3H into these elements has been made. The data obtained from autoradiographic study are discussed in relation to the hypothesis that the light cells are elements in active steroido-synthesis, the dark cells are resting elements, and the very dark cells are elements in degeneration.The authors wish to thank Mr. G. Gottardo for his technical assistance.