Protective Effect of Liv.52 on Alcohol-Induced Fetotoxicity

Gopumadhavan, S., Jagadeesh, S., Chauhan, B.L. and Kulkarni, R.D., R&D Center, The Himalaya Drug Company, Bangalore, India.


The adverse effects of maternal alcohol consumption on the development of the fetus are well known. The adverse effects of ethanol on the liver are now believed to be due to acetaldehyde formed as an intermediate metabolite of ethanol. Liv.52 has been shown to bring about faster elimination of acetaldehyde from the body and thus prevent alcoholic liver damage. Other toxic effects of alcohol may also be due to acetaldehyde and may be prevented by Liv.52. In this, study, rats were given 20% (v/v) ethanol in drinking water, during the gestation period, and the effect on maternal body weight and fetal outcome was noted. The protective effect of Liv.52 administration during the gestation period was studied. The results show that ethanol ingestion caused a decrease in gestational weight gain, total fetal weight, and number of live fetuses. There were increases in resorptions. Liv.52 administration reduced the deleterious effects of ethanol. The concentration of acetaldehyde in the amniotic fluid of ethanol-consuming animals was 0.727 µg/ml. Liv.52 administration lowered it to 0.244 µg/ml. The protective effect of Liv.52 could be due to the rapid elimination of acetaldehyde.

Key Words: Fetal Alcohol Syndrome, Acetaldehyde, Liv.52.

The adverse effects of maternal alcohol consumption of fetal development are well documented1-3. Even moderate drinking is clearly contraindicated during pregnancy. The resulting abnormality on the fetus consists of decelerated growth and a number of major and minor malformations. Stillbirths and spontaneous abortions are much more frequent in alcohol-imbibing pregnant women. Increased accumulation of acetaldehyde, an intermediate metabolite of ethanol, is believed to be an important factor for the adverse effects of ethanol4,5. Evaluating the embryotoxicity of two ethanol metabolites, Priscott6 reported that acetaldehyde in concentrations of 100 and 260 µM in the incubation medium had no deleterious effect on the gross morphology or viability of cultured 10-day old Albino Wistar rat embryos. However, at 800 µM concentration under similar conditions, it was overtly toxic causing rapid death. The other metabolite 2,3-butanediol, at a concentration of 25 µM, had no adverse effect6.

Liv.52 is a herbal formulation based on “AYURVEDA” and is known to protect the liver from damage induced by toxic substances, including alcohol, in experimental studies7. Liv.52 enhances the absorption of ethanol and rapidly reduces the acetaldehyde levels, which may explain its hepatoprotective effect on ethanol-induced liver damage8,9. Keeping these facts in mind, a study was undertaken to elucidate the protective effect, if any, of Liv.52 against the harmful effects of maternal alcohol consumption during the gestation period and also to determine the effect of Liv.52 on the acetaldehyde concentrations in the amniotic fluid of fetuses.


Albino rats of the Wistar strain (2.5-3.0 months old and weighing between 200-250 g) were used in the trial. These animals were given a standard diet (Hindustan Lever Pellets, Bangalore), clean tap water ad libitum, and kept in an air-conditioned room maintained at 22 ± 1°C with 60% relative humidity. All experiments were conducted between August and September, during which day and night periods are equal, and the animals were exposed to this natural day and night cycle. Assigned oral administrations in all the groups were conducted daily, between 9:00 and 11:00 a.m.

The vaginal smear of each rat was examined daily for 12 days for the degree of cornification of epithelium to select animals showing a regular estrous cycle of 5-6 days. In the third cycle, those female rats in proestrus were separated and allowed to mate with males of proven fertility by mass-mating technique. Female and male animals (ratio 1:2) were caged together overnight. The next morning, a vaginal smear from each rat prepared and examine under a microscope for spermatozoa. The positive females were marked and placed in individual cages and the day was termed as Day 1 of pregnancy.

The 27 rats selected by the above method were divided into three groups. Group 1 (n=5) served as negative control and received 10 ml/kg water once a day orally for 20 days, from Day 1 of gestation. Group 2 (n=12) served as positive control and received 20% v/v alcohol in water feeding bottles for 20 days from Day 1 of gestation, and 10 ml/kg water orally once daily. Group 3 (n=10) rats received Liv.52 (3g/kg/day) orally for 15 days before and during gestation, in addition to 20% v/v alcohol in drinking water during this gestation period. The quantity of water consumed with Liv.52 was the same as that of Group 1 and Group 2.

General observations of the mother rats were made in regard to any change in daily food and water intake, alcohol consumption, gestational body weight, behavior, and presence of vaginal bleeding.

On Day 20, 3 hours after the assigned drug administration, the animals were anesthetized with ether, the abdomen was opened to expose the uterus, and the number and placement of implantation sites, early and late resorptions, and live and dead fetuses were noted. The ovaries were removed and examined for the number of corpora lutea. Amniotic fluid collected and pooled from fetuses in each dam was subjected to acetaldehyde analysis by the head space gas chromatography method10. Due to the limitations of head space gas chromatography and the need to analyze samples soon after collection, estimation of acetaldehyde was conducted from five dams in each group.

The weights of all fetuses were recorded, and each fetus was examined from gross external variations. Half of the fetuses from each litter were examined for soft tissue malformations by using Wilson’s sectioning technique11. The remaining fetuses were examined for skeletal malformations by using the Alizarin Red stain technique12. Fertility and gestation indices were calculated by standard method13. All parameters were statistically determined by one-way analysis of variance with appropriate post-hoc analysis using SPSS/PC+ on an IBM-compatible AT 286.


The mean alcohol consumed per rat was 12.53 g/kg body weight/day in Group 2 (treatment alcohol + Liv.52). Two animals in Group 2 (treated with only alcohol) and one animal in Group 3 (alcohol + Liv.52-treated) did not show signs of pregnancy and were excluded on Day 4, thus leaving 10 and 9 animals, respectively, for evaluation. Tale 1 shows the mean alcohol consumption, maternal body weight gain, gestational food intake, and total number of live fetuses in each of the treatment groups. Twenty percent v/v alcohol ingestion in drinking water during the gestation period (Group 2) produced a significant reduction in the gestational weight gain and number of live fetuses as compared with the control group. Liv.52 treatment group showed no significant difference from the control group in these respects. The food intake was similar in all groups, and mean alcohol intake was not different between the alcohol alone and alcohol + Liv.52 groups. Figure 1 shows the mean litter weight of dams in the three groups. The mean litter weight was significantly lower in Group 2 treated with only alcohol as compared with the control group treated with tap water. The mean litter weight in the alcohol + Liv.52 group was not different from the control group, but was significantly higher than the alcohol alone group.

Figure 2 shows the mean of resorptions of dams in each group. The mean of resorptions in the female rats treated with alcohol alone was significantly higher as compared with the control group. In the Liv.52 + alcohol group the mean of resorptions was less than in the alcohol alone group, and the difference was statistically significant in spite of high variation in the alcohol alone group. The mean of resorptions in the Liv.52 + alcohol group was also significantly higher than in the control group.

The amniotic fluid of all fetuses from each dam was pooled for the estimation of acetaldehyde concentration. The mean results of five rats in each group treated with only alcohol and alcohol + Liv.52, respectively, are shown in Figure 3. The mean acetaldehyde concentrations in rats treated with only alcohol were significantly higher (0.727 ± 0.046) as compared with the alcohol + Liv.52-treated rats (0.244 ± 0.027).

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