Homeostasis changes induced by the action of ethanol on the materno-fetal complex in rats. VII. Effects on some alkaline and alkaline-earth metals of blood serum.

The effect of chronic biparental preconceptional alcoholization followed by alcohol consumption during the preimplantation period on the maternal and fetal homeostasis of some alkaline and alkaline-earth metals was studied in albino rats. Female and male rats consumed 20% ethanol (v/v) for 50-60 days before mating (group E). Further on pregnant females were alcoholized until day 5 of pregnancy. Control females and males consumed tap water (group C). Animals were sacrificed on day 20 of pregnancy. Blood samples were obtained from fetuses (the blood of each litter being pooled) and from pregnant females. Biochemical determinations were made by atomic emission spectroscopy for Na and K and by atomic absorption spectroscopy for Ca and Mg. Early and late mortality of conceptuses were checked. Non significant changes were observed within group E in the maternal serum as to Na (increase) and K(decrease), while Ca and Mg showed a significant decrease. Similar changes were found in the pooled fetal serum, the decrease of K and Mg being less marked. No structural anomalies were detected. With respect to tanatogenesis, both early resorptions and late fetal death were increased.

Homeostasis changes induced by the action of ethanol on the materno-fetal complex in rats. V. Late fetal effects of acute intoxication during the preimplantation period.

The late fetal effect of ethanol administered during the preimplantation period (in acute experiment) was investigated. Ethanol was injected i.v. (33.16% v/v in a.d., 4.80 ml/kg b.w.) to pregnant female rats on day 2 and 4 of pregnancy. Some effects concerning biochemical and morphological homeostasis on at-term fetuses (day 20 of pregnancy) were studied. Data obtained were compared with those of the control group. Biochemical investigation performed on hepatic DNA and on some serum metabolites revealed the following statistically non-significant changes: the increase of fetal hepatic DNA; the increase of total protein determined from pooled fetal serum of the whole litters; hypoalbuminemy and hyperglobulinemy; hypo-alpha 1-globulinemy and hyper-alpha 2-, beta-, gamma-globulinemy; the increase of total lipids, decrease of cholesterol; increase of uric acid and urea. In the amniotic fluid the following statistically non-significant values were found: increase of proteins, lipids, uric acid and urea content and decrease of cholesterol. Ponderal somatometry evidenced a statistically significant decrease of fetal and placental wet weight. The changes found show that--in our experimental conditions--the i.v. administration of ethanol during the preimplantation period does not significantly influence the late, fetal biochemical values and induces a significant lowering of fetal and placental wet weight and a significant increase of late fetal mortality.

Homeostasis changes induced by the action of ethanol on the materno-fetal complex in rats. IV. Late maternal effects following acute intoxication during the preimplantation period.

The possible late maternal effect on biochemical homeostasis of acute administration of ethanol during the preimplantation period was studied on pregnant albino female rats (Wistar strain). Females were injected i.v. with a 33.16% (v/v) solution of ethanol (4.80 ml/kg body weight) on day 2 and 4 of pregnancy, the animals being sacrificed on day 20. The effects on hepatic DNA biosynthesis and on some serum metabolites (proteins, lipids and some non-protein nitrogen compounds) were determined in the treated and in an untreated control group. In the treated group a non-significant increase of hepatic DNA concentration was found. Homeostasis changes of serum proteins involved: a slight increase of total serum protein content, hypoalbuminemia and hyperglobulinemia (non-significant values). Lipid metabolites showed also non-significant changes: increase of total lipids and decrease of cholesterol. Uric acid and urea (non-protein metabolites) concentration was increased. This increase, in spite of its significance level, must be taken into account and may be due to the presence of dead fetuses and to the consecutive renal lesions.

Effect of alcoholization upon the maternal and fetal hepatic DNA and the maternal serum-proteins in pregnant albino rats.

Experiments were performed on pregnant albino rats (Wistar strain) of 150-200 g b.w. Biochemical investigations involved the determination of maternal and fetal hepatic DNA content (Spirin's method), of maternal serum proteins (total protein content and electrophoretic fractions). The mean litter size, early resorptions, fetal mortality, and some biochemical data of fetuses were also determined. The main statements were as follows: The chronic, peroral administration of 20% ethanol to pregnant albino rats before and during pregnancy induced changes of the biosynthesis of hepatic DNA: a nonsignificant increase as compared with the control group was recorded. The control of serum proteins revealed an increase of the total protein content and of the total electrophoretic serumglobulin fractions. Within the globulin fraction, a hyper-alpha-globulinemia, and a hypo-beta and gamma globulinemia were detected. In the fetuses of the experimental group a slight but statistically significant increase of the hepatic DNA content appeared. In the experimental group the early resorption rate (10.97%) and the fetal mortality (2.43%) was increased in comparison with the control group (0%). In the alcoholized mothers a nonsignificant decrease of the crown-rump length and a significant decrease of fetal and placental weight could be observed.

The effect of ethanol upon early development in mice and rats. VI. In vivo effect of acetaldehyde upon preimplantation stages in rats.

In completion of the previously outlined "experimental alcohol blastopathy", the role of acetaldehyde in the induction of preimplantation pathological changes in rat embryos has been controlled. Two experimental models were used: the direct administration of acetaldehyde by gavage and the blockage of acetaldehyde metabolization by ANTALCOL (an aldehyde-dehydrogenase blocking compound). The main results were as follows: The exogenous acetaldehyde in the blood of pregnant animals has an obvious effect upon the developmental rate during the late preimplantation period (retarding segmentation, blastulation), and in one of the experimental models upon the oviductal-uterine migration rate. The increase of the blood acetaldehyde level by blockage of its further metabolization has a more marked effect as compared with the direct intravenous administration of the substance. According to our previous observations the intravenous application of ethanol on the same day (day 4) has no such effect. The direct noxious influence upon the developing preimplantation embryos (fragmentation) of the increased level of acetaldehyde obtained by ANTALCOL treatment is similar but more marked than this effect obtained previously by ethanol administration. The same effect observed after the direct administration of the substance is less marked than the effect of ANTALCOL treatment but more marked than the effect of intravenous ethanol administration. These results attest that acetaldehyde may contribute (alone or together with the effect of ethanol) to the induction of "experimental alcohol blastopathy". The less marked action of the substance proper introduced into the blood stream may be due--in our opinion--to its possible alteration during the period between distillation and application.

The presence of ethanol in the oviductal and uterine luminal fluids of alcoholized rats.

Blood ethanol level and the presence of ethanol in the oviductal and uterine luminal fluids were determined in female albino rats (Wistar strain) following chronic alcoholization by 20% ethanol offered ad libitum in drinking water during a period of 40-50 and 90-100 days, respectively. Ethanol was found both in the oviductal and in the uterine luminal fluids in lower concentrations as compared with the blood ethanol level. The findings presented suggest the possible direct noxious action of ethanol upon preimplantation development (effects detected in previous investigations). Problems raised by present results are discussed.

The effect of ethanol upon early development in mice and rats. I. In vivo effect upon preimplantation and early postimplantation stages.

The preimplantation and early postimplantation effect of chronic alcohol consumption (at least a month before mating and during pregnancy until killing) and of acute ethanol intoxication during the preimplantation period (i.v. infection of ethanol) was studied on albino rats (Wistar) and albino mice (RAP). The main results were as follows: Chronic alcoholization. Rats: significant retardation of preimplantation development and in early postimplantation stages; a tendency of lowering of the mean litter size. Mice: significant increase of the number of preimplantation pathological forms; a tendency of lowering of the mean litter size. Pathological changes show, both in rats and mice, an obvious "litter effect". Acute ethanol intoxication. Rats: significant retardation in some litters, normal or even advanced development in others. This effect differs from the previously reported effect of acute ethanol intoxication during early postimplantation stages. The results obtained attest the prenatal noxious effect of chronic ethanol consumption in both species used and of acute ethanol intoxication during preimplantation development upon early postimplantation development in rats. Within the limits of extrapolation possibilities, they represent a risk signal for other species (including human).