RESUMEN
Twenty years ago, we detected the interdependence between structure and function of rat liver Golgi complexes that are characteristic for streptozotocin diabetes, which served us in further investigations as a useful indicator of the effectiveness of drugs we were testing. This work presented results obtained in eight groups of rats (four control and four diabetic) that were administered orally either bis(maltolato)oxovanadium(IV) [BMOV] or maltol alone. The activities of the rat liver Golgi marker enzyme, galactosyltransferase [GalT], as well as the morphology of Golgi complexes were studied in situ using an electron microscope; parallel estimations of vanadium concentration and phospholipid percentage were made in Golgi-rich preparations isolated from the liver. Our main findings were normalization in diabetic animals orally treated with 1.8 mmol BMOV in 0.09 mol NaCl solutions over seven days, which demonstrated an accompanying increase in phosphatidic acid (PA) percentage (p < 0.05) compared to controls. In the diabetic groups, Pearson's test showed a positive double correlation between GalT activity, vanadium concentration, and PA percentage in Golgi-rich membrane preparations from the liver. Additionally, a negative correlation was found between vanadium concentration and phosphatidylcholine percentage in the fractions.
Asunto(s)
Diabetes Mellitus Experimental/patología , Aparato de Golgi/efectos de los fármacos , Aparato de Golgi/ultraestructura , Hipoglucemiantes/farmacología , Hígado/efectos de los fármacos , Hígado/patología , Pironas/farmacología , Vanadatos/farmacología , Animales , Diabetes Mellitus Experimental/enzimología , Diabetes Mellitus Experimental/metabolismo , Femenino , Galactosiltransferasas/metabolismo , Aparato de Golgi/enzimología , Aparato de Golgi/metabolismo , Hígado/enzimología , Hígado/metabolismo , Microscopía Electrónica , Concentración Osmolar , Fosfolípidos/metabolismo , Ratas , Ratas Wistar , Cloruro de Sodio/farmacología , Vanadio/farmacocinéticaRESUMEN
Among the previously studied organic vanadium derivatives showing an anti-diabetic action, we investigated a new complex, bis(2,2'-bipyridine)oxovanadium(IV) sulphate. We tested its ability to normalise parameters previously described for streptozotocin (STZ)-diabetes, such as lower yields of Golgi-rich membrane fraction isolation, decreased activity of Golgi membrane marker enzyme - galactosyltransferase (GalT) - and altered morphology of rat liver Golgi complexes. Oral application as a drinking solution of 1.8 mmol bis(2,2'-bipyridine)oxovanadium(IV) (dissolved in 0.09 M NaCl) caused a similar dispersion of GalT activities in both vanadium treated groups, control and diabetic. Very low activities of the enzyme (characteristic for untreated diabetes) we found only in approximately 35 % of the STZ-diabetic rats treated with the new vanadium compound. The morphology of liver Golgi complexes in diabetic rats treated with bis(2,2'-bipyridine)oxovanadium(IV) sulphate was improved, which manifested itself in the reappearance of vacuoles with VLDL and coated and uncoated secretory vesicles. In view of biochemical and morphological parameters of normalised diabetic rat liver Golgi apparatus, the new vanadium complex was more effective than bis(oxalato)oxovanadium(IV) or bis(kojato)oxovanadium(IV), but in our experimental model, the best anti-diabetic, orally applied drug was the bis(maltolato)oxovanadium(IV) previously investigated.
Asunto(s)
2,2'-Dipiridil/farmacología , Diabetes Mellitus Experimental/tratamiento farmacológico , Aparato de Golgi/efectos de los fármacos , Hipoglucemiantes/farmacología , Hígado/efectos de los fármacos , Compuestos Organometálicos/farmacología , Animales , Glucemia/metabolismo , Peso Corporal , Diabetes Mellitus Experimental/metabolismo , Femenino , Galactosiltransferasas/metabolismo , Aparato de Golgi/metabolismo , Aparato de Golgi/ultraestructura , Hígado/enzimología , Hígado/metabolismo , Hígado/ultraestructura , Microscopía Electrónica , Ratas , Ratas WistarRESUMEN
Oral treatment with maltol or bis(maltolato)oxovanadium(IV) [BMOV] alters the biochemical activity of the rat liver Golgi marker enzyme, i.e., galactosyltransferase (GalT), and the organelle morphology in a relatively short time. Four groups of rats were investigated: control (C), treated with BMOV for 2 days (pVC), treated with BMOV for 7 days (C+V), and treated with maltol alone for 7 days (C+M). All drugs were administered as drinking solutions. These conditions were used, because normalization of galactosyltransferase activity (GalT) and morphology of rat liver Golgi complexes were previously found by us in streptozotocin-induced diabetes. In this paper, we present the influence of BMOV or maltol alone (as a vanadium ligand in BMOV compound) on rat liver Golgi complexes. The lowest statistically significant enzyme activity, in comparison with three other groups of rats (p < 0.01), was found in rats treated with BMOV solution for two days (pVC). Liver Golgi complexes in these rats showed relatively slight changes as compared with controls. The activity of GalT was similar to controls of the C+V and C+M groups. Morphological examinations of the Golgi apparatus in rats treated with vanadium salts revealed a slightly increased secretory activity. In response to various agents used in experiments, the Golgi complexes were generally reduced in size, except for the (C+M) group. Not only cisternae, but also vacuoles and associated vesicles on both sides of stacks were reduced in almost all Golgi structures. Ultrastructural findings were generally in agreement (except for pVC group) with biochemical results (yields of liver Golgi-rich fractions, activity of galactosyltransferase) obtained in the same rats.
Asunto(s)
Aparato de Golgi/metabolismo , Aparato de Golgi/ultraestructura , Hígado/metabolismo , Hígado/ultraestructura , Pironas/farmacología , Vanadatos/farmacología , Administración Oral , Animales , Femenino , Ratas , Ratas WistarRESUMEN
In comparison with untreated control, reduced body and liver weights were found in two groups of rats (such as control and STZ-diabetic) treated orally with bis(kojato)oxovanadium(IV) solution. Free blood sugar in STZ-diabetic rats was lower by about 38%, but did not achieve euglycemic values. Yields of Golgi-rich fraction were lower than those in untreated controls, similar to the activity of galactosyl transferase (GalT) in both vanadium treated groups (control and diabetic). Under electron microscope in the control kojate-treated group, subcellular changes were observed. The morphology of Golgi apparatus was typical, resembled that in untreated animals. In diabetic animals treated with kojate subcellular changes were less severe. Golgi apparatus was usually semicircular or arched in shape similar to that observed previously in diabetic untreated rats.
Asunto(s)
Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Hígado/efectos de los fármacos , Compuestos Organometálicos/uso terapéutico , Pironas/uso terapéutico , Administración Oral , Animales , Biopsia , Glucemia/análisis , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Experimental/patología , Femenino , Galactosiltransferasas/análisis , Aparato de Golgi/efectos de los fármacos , Aparato de Golgi/ultraestructura , Hígado/enzimología , Hígado/patología , Microscopía Electrónica , Ratas , Ratas Wistar , EstreptozocinaRESUMEN
In this study oral treatment with bis(kojato)oxovanadium(IV) solution was administered twice. The first, short vanadium treatment (so called pretreatment) was used to accustom animals to the flavour of this liquid. After 2-2.5 weeks the second treatment, in high concentration and for a longer time served as a "drug". The physiological parameters such as weights of animals and their livers, reduction of liquid and food intake were lower than in control. Activity of Golgi marker enzyme GalT was significantly lower in both vanadium treated groups (p < 0.01), but in diabetic vanadium treated group it was higher, albeit not significantly, than in control vanadium treated rats. The morphology of Golgi complexes in diabetic rats on prolonged vanadium treatment was similar to that in the one week treated rats (see Part I). Rounded stacks of cisternae characteristic of untreated streptozotocin (STZ)-diabetes were also seen in vanadium treated diabetic rats, but in comparison with untreated diabetic livers, the secretory activities of Golgi complexes were preserved or even stimulated.
Asunto(s)
Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Hígado/efectos de los fármacos , Compuestos Organometálicos/uso terapéutico , Pironas/uso terapéutico , Administración Oral , Animales , Biopsia , Glucemia/análisis , Diabetes Mellitus Experimental/patología , Galactosiltransferasas/análisis , Aparato de Golgi/efectos de los fármacos , Aparato de Golgi/ultraestructura , Hígado/enzimología , Hígado/patología , Microscopía Electrónica , Ratas , Ratas Wistar , Estreptozocina , Factores de TiempoRESUMEN
(1) Both vanadyl oxalate and streptozotocin (STZ) caused in comparison with untreated control statistically significant increase (P<0.001 and P<0.02) of PLs (micromoles of P(i) per mg of protein) in rat liver Golgi-rich membrane fraction. (2) The diabetic, vanadium treated rats (D+V) showed lower than control-treated (C+V) content of PLs in these fractions. (3) Three experimental groups of rats: control-treated (P<0.01), diabetic treated with vanadium (P<0.05) and untreated diabetic (P<0.02), had a higher percentage of PI (phosphatidylinositol) in comparison with untreated-control animals.
Asunto(s)
Diabetes Mellitus Experimental/metabolismo , Aparato de Golgi/metabolismo , Hígado/metabolismo , Oxalatos/farmacología , Fosfolípidos/metabolismo , Vanadatos/farmacología , Animales , Biomarcadores , Femenino , Galactosiltransferasas/metabolismo , Aparato de Golgi/efectos de los fármacos , Técnicas In Vitro , Hígado/efectos de los fármacos , Membranas/efectos de los fármacos , Membranas/metabolismo , Ratas , Ratas WistarRESUMEN
The relation between bis(maltolato)oxovanadium(IV) (BMOV) influencing the biochemical activity of rat liver Golgi apparatus and the morphology of this organelle was studied in normal and streptozotocin-diabetic rat livers. Ultrastructural examinations revealed marked differences in the morphology of Golgi apparatus in three groups of animals. In the control rats treated only with 0.5% NaCl we did not find any biochemical and morphological changes. Marked changes were found in the rat liver after 1.8 mmol BMOV in 0.5% NaCl (as drinking solution) applied for 7 days, so-called "control" group for vanadium. In this group Golgi apparatus seemed shorter than in the diabetic animals. Finally, the same treatment of rats with previously induced SZ-diabetes, showed relatively small morphological alterations. The ultrastructural observation was compatible with the activity of galactosyltransferase (GalT), the Golgi marker enzyme. In diabetic rats treated with BMOV the activity of this enzyme was almost the same as in controls. Summing up dramatic alterations, previously found in diabetic-untreated rats [22], normalized after orally applied BMOV solution, even after a short time.
Asunto(s)
Diabetes Mellitus Experimental/enzimología , Diabetes Mellitus Experimental/patología , Inhibidores Enzimáticos/farmacología , Galactosiltransferasas/efectos de los fármacos , Aparato de Golgi/efectos de los fármacos , Hipoglucemiantes/farmacología , Hígado/ultraestructura , Pironas/farmacología , Vanadatos/farmacología , Animales , Femenino , Galactosiltransferasas/metabolismo , Aparato de Golgi/enzimología , Aparato de Golgi/ultraestructura , Microscopía Electrónica , Ratas , Ratas Wistar , EstreptozocinaRESUMEN
Recent studies have shown the insulin-like effect of vanadyl sulphate or sodium ortho (or meta-)vanadate administered orally to rats. Toxicity of these drugs and reluctance by the animals to drink the solutions and take food, concerning the amelioration of some diabetes syndrome discussed in 1994 by Domingo et al. (1), McNeill et al. (2) and Wiliams and Malabu (3), prompted us to investigate a new vanadate complex: disodium bis(oxalato)oxovanadate (IV), Na2[VO(OX)2]H2O. The main object of the experiment was to study whether this complex administered as 3 mmol/l solution in 0.5% NaCl during 7 days could act on the subcellular level and influence the activity of liver Golgi membrane galactosyltransferase activity. Free blood sugar level was lowered (but was still higher than in the control group) in diabetic rats after seven days of vanadate action and was accompanied by lowered, however not statistically significant, serum triglyceride levels. The yields of isolated Golgi-rich membrane fractions were about half of the level in diabetic groups (untreated and treated with vanadium) compared with the control groups. Purity of these membrane fractions, expressed as nmol Gal transferred per mg of proteins and per h, was the same in four groups investigated and showed the possibility to compare them. Activity of galactosyltransferase calculated in nmol Gal transferred per 1 g of liver and per 1 h or per whole liver in the same time (as a possibility of glycosylation of the secretory and membrane glycoproteins) was lower in both diabetic groups. However, after vanadium treatment (D+V group), the activity was higher than in untreated diabetic rats (D group) in three of five investigated animals. Vanadyl-oxalate complex did not normalize in a statistically significant manner the enzyme activity which was significantly lower in diabetes than in control. This is similar to insulin influence on the galactosyltransferase activity reported previously by Kaczmarski et al. in 1981 (4) and Kordowiak et al. in 1981 (5).
Asunto(s)
Galactosiltransferasas/metabolismo , Aparato de Golgi/ultraestructura , Insulina/farmacología , Membranas Intracelulares/enzimología , Hígado/ultraestructura , Oxalatos/farmacología , Vanadatos/farmacología , Vanadio/farmacología , Animales , Glucemia/metabolismo , Diabetes Mellitus Experimental/enzimología , Femenino , Galactosa/metabolismo , Aparato de Golgi/efectos de los fármacos , Aparato de Golgi/enzimología , Membranas Intracelulares/efectos de los fármacos , Ratas , Ratas WistarRESUMEN
This paper presents yields of Golgi-rich membrane isolation, the activity of galactosyltransferase (GalT), the marker enzyme of Golgi apparatus as well as the morphology of the organelle from the livers in situ, in two groups of rats. One group consisted of control rats injected twice intraperitoneally with LEPK. Second group consisted of rats injected with LEPK and additionally after 24hrs given streptozotocin (SZ) to induce experimental diabetes. The results were compared with our previous investigations in control and diabetic rats. In the latter the activity of GalT was diminished, therefore diminishing glycosylation ability, and destructing Golgi apparatus morphology. This experiment shows that two-fold injection of LEPK prior to SZ does not prevent from changes in such biochemical parameters as free blood glucose level, yield of liver Golgi membranes isolation or total activity of GalT. For the first time in c. 30% of investigated rats the inactive enzyme of Golgi apparatus was found in the rats treated with LEPK+SZ. Morphological investigations of liver Golgi apparatus in rats treated with LEPK show slightly increased secretory activity with similar to untreated control rats morphological structure of this organelle. In the rats treated with LEPK and SZ the same morphological changes as in diabetic liver were found, however, such dramatic alterations as in SZ-diabetic rats were never found, irrespective of active or inactive GalT.
Asunto(s)
Diabetes Mellitus Experimental/metabolismo , Aparato de Golgi/metabolismo , Hígado/metabolismo , Extractos Vegetales/farmacología , Polen/química , Animales , Glucemia/efectos de los fármacos , Diabetes Mellitus Experimental/patología , Femenino , Galactosiltransferasas/metabolismo , Aparato de Golgi/efectos de los fármacos , Aparato de Golgi/ultraestructura , Hígado/efectos de los fármacos , Hígado/ultraestructura , Tamaño de los Órganos/efectos de los fármacos , Ratas , Ratas WistarRESUMEN
Addition in vitro of ethanol solution of PGE1 to isolated Golgi-rich membrane fraction caused great alterations in galactosyltransferase activity, marker enzyme of these membranes. Ethanol as a solvent of PGE1 has an influence on the activity of the enzyme as well as the membrane permeability, different drugs penetration and availability of substrates. Then, additional control with ethanol in identical concentration as in the investigated sample was performed. In a dose 1 microgram PGE1 per 1 mg of protein (and lower concentration of ethanol c. 0.09%) the stimulation of this enzyme activity (excluding 2 and 6 hours s after Golgi membrane isolation) was above 30% in comparison with the control.
Asunto(s)
Alprostadil/farmacología , Etanol/farmacología , Aparato de Golgi/enzimología , Hígado/enzimología , N-Acetil-Lactosamina Sintasa/metabolismo , Animales , Permeabilidad de la Membrana Celular/efectos de los fármacos , Femenino , Aparato de Golgi/efectos de los fármacos , Técnicas In Vitro , Metabolismo de los Lípidos , Fosfolípidos/metabolismo , Proteínas/metabolismo , Ratas , Ratas Wistar , SolventesRESUMEN
Nocardia lysozyme digest (NLD), a particulate fraction from Nocardia opaca, is able to induce antitumor activity to SaL-1 tumor cells (lung sarcoma) in Balb/c mice. In mice immunized with NLD inhibition of tumor growth and prolonged survival of tumor bearing animals was observed. Macrophages isolated from peritoneal cavity and stimulated with NLD release a few arachidonic acid metabolites, mostly PGE 2. Macrophages from tumor bearing mice are more sensitive to Nocardia antigens than normal. Both in vitro and in vivo experiments have documented that Nocardia is an active immunomodulator.
Asunto(s)
Adyuvantes Inmunológicos , Inmunización , Neoplasias Pulmonares/prevención & control , Nocardia/inmunología , Animales , Neoplasias Pulmonares/mortalidad , Ratones , Ratones Endogámicos BALB C , Muramidasa/farmacología , Células Tumorales CultivadasRESUMEN
In vitro addition of 16,16'-dimethyl prostaglandin E2 to Golgi-rich membrane fraction in final concentration of 0.1 microgram/1 mg of protein increased generally the activity of galactosyltransferase in comparison with control. The percentage of phospholipids in the whole fraction was similar in both investigated groups, only the sum of phosphatidylethanolamine+phosphatidic acid was significantly lower after addition of dmPGE2 than in the control (0.001 < P < 0.01).
Asunto(s)
16,16-Dimetilprostaglandina E2/farmacología , Galactosiltransferasas/metabolismo , Hígado/efectos de los fármacos , Hígado/enzimología , Animales , Femenino , Aparato de Golgi/enzimología , Aparato de Golgi/metabolismo , Técnicas In Vitro , Hígado/metabolismo , N-Acetil-Lactosamina Sintasa/metabolismo , Fosfolípidos/metabolismo , Ratas , Ratas WistarRESUMEN
The mobility of 5-doxylstearic acid spin label (5-SASL) in the intact rat liver Golgi membranes of streptozotocin diabetes was studied as a function of free blood sugar level and temperature. During development of diabetes, indicated by the increase of the free blood sugar level, the membrane fluidity measured in the physiological temperature range (1) does not change in comparison with control in light diabetes, (2) decreases significantly in advanced diabetes and (3) again increases to the control level in heavy diabetes (the free blood sugar levels being 200-250 mg/100 ml, 250-350 mg/100 ml and greater than 350 mg/100 ml, respectively). The development of streptozotocin diabetes is accompanied by significant changes in lipid composition of liver Golgi membranes as also shown in our previous observations. The measurements of motion of 5-SASL in Golgi membranes as well as in vesicles, made from commercially available lipids of composition close to the liver Golgi membranes, show that a decrease of cholesterol contents is the main factor which induces the increase membrane fluidity. We suggest that in the heavy diabetes the hemostatic regulation in the lipid composition leads to minimization of alterations in membrane fluidity to obtain comparatively normal activity of certain membrane enzymes.
Asunto(s)
Diabetes Mellitus Experimental/fisiopatología , Aparato de Golgi/ultraestructura , Membranas Intracelulares/fisiología , Hígado/ultraestructura , Fluidez de la Membrana , Animales , Glucemia/metabolismo , Óxidos N-Cíclicos , Espectroscopía de Resonancia por Spin del Electrón , Femenino , Liposomas , Ratas , Ratas Endogámicas , Marcadores de Spin , TemperaturaRESUMEN
In 10-11 days after single, intraperitoneal injection of streptozotocin we found a lower (although not statistically significant) yield of Golgi-rich membrane fraction in comparison with control. In these rats similar to control specific activity of galactosyltransferase in nM Gal transferred per h and mg of protein, and statistically significant lower total activity of this enzyme expressed in nM Gal transferred per h and per total liver (t = 2.9666, 0.01 less than p less than 0.05) were found. Organization of Golgi apparatus in situ showed alterations that indicated suppression of secretory activity. Within the dm PGE2 protected animal group the obtained data equal control data.
Asunto(s)
16,16-Dimetilprostaglandina E2/farmacología , Crioprotectores , Aparato de Golgi/efectos de los fármacos , Hígado/efectos de los fármacos , Prostaglandinas E Sintéticas/farmacología , Estreptozocina/farmacología , 16,16-Dimetilprostaglandina E2/metabolismo , Animales , Femenino , Congelación , Galactosiltransferasas/metabolismo , Aparato de Golgi/enzimología , Aparato de Golgi/ultraestructura , Hígado/ultraestructura , Microscopía Electrónica , Ratas , Ratas Endogámicas , Fracciones SubcelularesRESUMEN
16,16'-Dimethylprostaglandin E2 was administered to rats in three doses: 30 min prior and 24 h and 48 h after a single intraperitoneal injection of streptozotocin. The Golgi membrane fraction was analyzed 6 days after streptozotocin injection. It has been found that prostaglandin restores the Golgi membrane fraction and the activity of UDP-Gal----GlcNAc transferase, both significantly decreased upon treatment with streptozotocin alone. Morphology of the liver Golgi apparatus studied by the electron microscopy was similar to that of control from untreated rats although streptozotocin alone significantly decreased the size of this organelle.
Asunto(s)
16,16-Dimetilprostaglandina E2/farmacología , Hígado/efectos de los fármacos , Prostaglandinas E Sintéticas/farmacología , Estreptozocina/antagonistas & inhibidores , Animales , Femenino , Hígado/enzimología , Hígado/ultraestructura , N-Acetil-Lactosamina Sintasa/antagonistas & inhibidores , N-Acetil-Lactosamina Sintasa/metabolismo , Ratas , Ratas Endogámicas , Estreptozocina/toxicidadRESUMEN
After dmPGE2 treatment, both alone or together with streptozotocin, the lower, statistically significant total PL contents (in mumoles P per mg of protein) in rat liver Golgi-rich membrane fractions were found. In these groups of investigated rats, the percentage of PE+PA were statistically significantly lower than in control. For the present it is impossible to certify, if there is a causal-result dependence between these two facts. The administration of streptozotocin alone (both after 6 or 11 days) caused the increase of PE+PA percentage, however not statistically significant.