In three experiments (2 on famales,l on males),we determined the blood flow in the tibia and the distal part of the femur, together with cardiac output (by meansi of 85Sr-microspheres),tibial bone density and tibial ash weight related to bone volume. We found that 1) the bone blood flow always fell significantly after oestradiol benzoate, 2) no change occurred after norethisterone in doses corresponding to those of oestradiol benzoate, but the blood flow showed a tendency to fall after doses one order higher (it decreased significantly in one case only), 3) the density of the tibia and tibial ash weight related to bone volume rose nonsignificantly after oestradiol benzoate, but fell (mostly statistically significantly) after norethisterone. The lowering of the bone mineral indexes in rat bones after norethisterone is a surprising and potentially significant finding requiring further verification.
We studied the effects of hydrocortisone as a possible regulatory factor of bone blood flow and metabolism. Local bone blood flow in the tibia, distal femur, lumbar vertebra and some soft tissues (using 85Sr-microspheres), as well as 45Ca and 3H-proline incorporation into the tibia, bone density and ash weight per ml of the tibia were measured in sham-operated and oophorectomized female rats in which the influence of hydrocortisone administration (0.004 % diet for 5 weeks) was followed. Hydrocortisone markedly lowered 85Sr-microsphere uptake and blood flow through the bones of non-castrated female rats as well as elevated circulatory values in oophorectomized rats. The changes were nearly identical in the three bone samples measured; among the soft tissues only the kidneys showed a less pronounced decrease. Circulatory changes in the bones seem to be caused by local vascular reactions. Hydrocortisone also lowered the 24-hour incorporation of 45Ca and 3H-proline into the tibia of both non-castrated and oophorectomized females. In the tibia of oophorectomized rats, hydrocortisone normalized the decreased bone density and ash weight. The adrenocortical hormones are known to block eicosanoid synthesis by the inhibition of arachidonic acid production. It is possible, therefore, that local circulatory changes in the bones of rats, induced by hydrocortisone, are mediated by the changes in prostaglandin production.
An increase in bone blood flow (BBF) was observed in rats after castration whereas a decrease in BBF occurred after oestradiol or testosterone. The possible participation of prostaglandins in these changes was demonstrated. The present results show that the endothelium-derived relaxing factor, i. e. nitric oxide (EDRF-NO), might play a role in these hormonal actions on BBF. Until now, almost nothing is known about the possible action of NO on bone circulation. Methylene blue (MB) as a substance blocking EDRF-NO was administered to sham-operated or oophorectomized (OOX) female rats. We determined local blood flow (85Sr-microsphere uptake), cardiac output, blood pressure, heart rate, density of the tibia and ash weight, as well as 24-h incorporation of 45Ca and 3H-proline into the tibia. The administration of MB (0.5 % in the food for 4 weeks) significantly lowered both 85Sr- microsphere uptake and blood flow values in the tibia and distal femur of sham-operated and OOX rats. MB lowered cardiac output and blood pressure to the same extent, indicating no change in the vascular resistance. After the administration of MB (0.1 % in the food), 85Sr-microsphere uptake decreased significantly in the tibia of OOX females while no significant change was found in soft tissues. Bone density and ash weight were significantly lower in OOX rats and in sham-operated rats after MB treatment. Finally, the 24-h incorporation of both 45Ca and 3H-proline decreased significantly in OOX females after MB administration (0.04 % in the food). It can be concluded that 1) MB lowers BBF, suggesting the participation of EDRF-NO in BBF regulation, 2) MB does not influence or may even suppress cardiac output and blood pressure in high dosage, 3) MB lowers 24-hour incorporation of 45Ca and 3H-proline into the tibia of OOX rats, which is in agreement with the circulatory effect, 4) MB lowers bone density and ash weight of the tibia in non-castrated female rats. The effects of MB observed in our experiments partially differ from those of arginine-derived blocking agents. This requires further elucidation.