RESUMEN
Two enzymes with tonin-like activity, designated rSMT3 and rSMT4, were purified from rat submandibular glands and another, rPT1, was obtained from the prostate. The three enzyme fractions hydrolysed angiotensin I, angiotensinogen (AG) and synthetic AG(1-14) to form angiotensin II. With angiotensin I as substrate, pH optima were 6.5 for rSMT3, 6.8 for rSMT4 and 7.5 for rPT1. With AG(1-14), the three enzymes had optimal activity at pH 7.5. The three enzymes had negligible activity upon a kallikrein substrate, Ac-Phe-Arg-Nan. The enzymes were inhibited by aprotinin, soybean trypsin inhibitor and phenylmethanesulfonyl fluoride but not by two angiotensin converting enzyme inhibitors, ethylenediaminetetracetic acid or enalaprilat. N-tosyl-L-phenylalanine chloromethyl ketone (1 mM) inhibited rPT1 and rSMT4 but not rSMT3. Molecular weights (SDS-PAGE) were 31,700 for rSMT3, 29,800 for rSMT4 and 28,100 for rPT1. Total activity in the prostate is 150-times lower than in the submandibular gland, where 92% of the tonin activity is related to rSMT4. Physical and chemical properties suggest that rSMT4 is tonin, whereas rSMT3 and rPT1 are tonin-like enzymes which can generate angiotensin II from different substrates.
Asunto(s)
Peptidil-Dipeptidasa A/metabolismo , Próstata/enzimología , Serina Endopeptidasas/aislamiento & purificación , Glándula Submandibular/enzimología , Angiotensina I/metabolismo , Angiotensina II/metabolismo , Angiotensinógeno/metabolismo , Animales , Electroforesis en Gel de Poliacrilamida , Concentración de Iones de Hidrógeno , Calicreínas/metabolismo , Masculino , Ratas , Ratas Endogámicas , Serina Endopeptidasas/metabolismo , Inhibidores de Serina ProteinasaRESUMEN
Groups of animals (Wistar rats) were fed with rations doped with uranyl nitrate at concentrations ranging from 0.5 to 100 ppm. The uranium content in the ashes of the organs was measured by the neutron-fission track counting technique. The most striking result is that the transfer coefficients, as a function of the uranium concentration, exhibit a concave shape with a minimum around 20 ppm-U for all organs. Explanations to interpret this finding are tentatively given.
Asunto(s)
Análisis de los Alimentos , Uranio/farmacocinética , Animales , Masculino , Neutrones , Fisión Nuclear , Ratas , Ratas Wistar , Distribución TisularRESUMEN
The regulation of mitochondrial energy metabolism plays an essential role in the central nervous system (CNS). Abnormalities of the mitochondrial respiratory chain often accompany neurodegenerative diseases. This makes mitochondria a perfect target for strategies of cellular protection against toxic compounds and pathological conditions. Steroid hormones, such as oestrogen, are well-known to fulfil a protective role in the brain during ischaemic and degenerative processes. Because astrocytes function as the major energy supplier in the CNS, we have analysed oestrogen effects on the mitochondrial respiratory chain of this cell type. In our studies, we applied semi- and quantitative polymerase chain reaction analysis of gene expression and polarographic measurements of the respiratory chain activity of mitochondria. We observed that structural and functional properties were regulated dependent on the oestrogen exposure time and the brain region, but independent of the nuclear oestrogen receptors. We could demonstrate that long-term oestrogen exposure increases the subunit gene expression of respiratory chain complexes and the mitochondrial DNA content, thereby indicating an up-regulation of the amount of mitochondria per cell together with an increase of mitochondrial energy production. This could represent an important indirect mechanism by which long-term oestrogen exposure protects neurones from cell death under neurotoxic conditions. On the other hand, we observed short-term effects of oestrogen on the activity of mitochondrial, proton-pumping respiratory chain complexes. In astrocytes from the cortex, respiratory chain activity was decreased, whereas it was increased in astrocytes from the mesencephalon. An increased production of reactive oxygen species would be the consequence of an increased respiratory chain activity in mesencephalic astrocytes. This could explain the different efficiencies of oestrogen-mediated short-term protection in distinct brain regions, but also indicates the limitations for a therapeutic short-term application of oestrogen.
Asunto(s)
Astrocitos/efectos de los fármacos , Corteza Cerebral/efectos de los fármacos , Transporte de Electrón/efectos de los fármacos , Estrógenos/farmacología , Genes Mitocondriales/efectos de los fármacos , Mesencéfalo/efectos de los fármacos , Animales , Astrocitos/metabolismo , Dominio Catalítico/efectos de los fármacos , Dominio Catalítico/fisiología , Células Cultivadas , Corteza Cerebral/metabolismo , Transporte de Electrón/fisiología , Regulación de la Expresión Génica/efectos de los fármacos , Mesencéfalo/metabolismo , Ratones , Ratones Endogámicos BALB C , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/fisiologíaRESUMEN
rSMT3, a tonin like angiotensin II generating enzyme present in rSMG presents a potent oxytocic effect on the isolated rat uterus, which was blocked by a B2 bradykinin receptor antagonist. Under optimal conditions of pH, rSMT3 liberates kinin at rate 19-fold greater than angiotensin II.