Rat Fetal Development Research Articles

Developmental Toxicity of Ethanolic Extracts of Leaves of Achyranthes aspera, Amaranthaceae in Rat Embryos and Fetuses.

IntroductionAchyranthes aspera is one of the medicinal plants widely used for fertility control in the local health system of Ethiopia.ObjectiveAssessment of developmental toxicity of ethanolic extracts of leaves of Achyranthes aspera in rat embryos and fetuses.MethodsFresh leaves were extracted by mixing the grinded powder with 70% ethanol. Then, the extract was given orally to gravid Wistar rats at doses of 250 mg/kg, 500 mg/kg and 1000 mg/kg from day 6–12 of gestation. On gestational days 12 and 20, embryos and fetuses were examined for developmental and gross malformations.ResultsOn day 12 embryos, the number of implantation sites and somites in 1000 mg/kg treated rats were significantly reduced. The number of implantation sites in pair-fed control and 1000 mg/kg groups was 11.2±0.86 and 8.34±0.65, respectively. Retarded development of hindlimb, forelimb, optic and olfactory systems was detected at a high dose. In addition, the number of branchial bar was significantly reduced in 1000 mg/kg dose. In near-term fetuses, significant reduction of litter weight and crown–rump length was seen at 1000 mg/kg dose. Crown–rump length in pair-fed control and 1000 mg/kg treated groups was 2.82±0.17 cm and 2.31±0.11 cm, respectively. Fetal resorptions and deaths in 1000 mg/kg were 1.45±0.65 and 0.81±0.67, respectively. However, external anomalies were not detected for all offspring at all doses.ConclusionThe finding suggests that ethanolic leaf extracts of A. aspera have detrimental effects on the development of rat embryos and fetuses at a higher dose. The possible teratogenic effects were indicated with the substantial retardation in embryonic and fetal development, decrease in number of implantation sites and rise in fetal resorptions and death. Moreover, it resulted in significant reduction in litter weight and crown–rump length at a higher dose.

Can melatonin correct the negative effect of experimental diabetes created during the maternal period on fetal rat development and puppies cognitive functions?

Can melatonin correct the negative effect of experimental diabetes created during the maternal period on fetal rat development and puppies cognitive functions?

Role of maternal nutritional supplementation on the hormonal profile and immunohistochemical analysis of testicular development of fetal rats

Introduction: This study elucidated the role of maternal microelement supplementation on fetal testicular development. Materials and Methods: Twenty-eight Wistar rats (21 females and 7 males) were randomly divided into seven groups comprising three females to one male: Group A – standard feed and water; Group B – diet deficient in all the micronutrients (Fe, Cu, Se) and water; Group C – diet fortified with 0.2 mg/kg of selenium and water; Group D – diet enriched with 40 mg/kg of iron and water; Group E – diet fortified with 8 mg/kg of copper and water; and Group F – diet enriched with all nutrients and water. All administration was via oral gavage; thereafter, animals were sacrificed at day 20 of pregnancy. Placenta measurements, testes, and blood serum were obtained for analysis. Results: The results showed statically a significant decrease (p<0.05) in placental and fetal weight (WPF), the distance between fetus and mother (DFM), the concentration of trace elements; FSH and LH levels among chaff only group when compared to the Control group. Testicular histomorphology and immunohistochemical studies of the animals in the chaff alone diet showed mild fetal Leydig cells in the interstitium, primitive germ cells in the testicular cord, pre-Sertoli cell necrosis, and decreased positive expression compared with the control. Conclusion: Administration of single and combined doses of nutritional supplements diet significantly preserved the fetal parameters, hormone profile, and histochemical analysis of the testis.

Folic acid rescues all-trans retinoic acid-induced anorectal malformations in rats.

To investigate whether folic acid (FA) can rescue anorectal malformations (ARMs) induced by all-trans retinoic acid (ATRA) in rats. Pregnant Sprague-Dawley rats were randomly divided into three groups. In the model group, rats were administered ATRA (110 mg/kg) by gavage on the 10th day of gestation (E10.5). Rats in the rescue group were administered FA (40 mg/kg) by gavage 0.5 and 12 hr after treatment with ATRA. All fetuses were harvested on E20.5 using cesarean section and examined for ARMs. We recorded the weight, body length, tail length, and associated malformations of all the embryos. Hematoxylin and eosin staining was used to analyze the histopathology of the fetuses. Control rats did not show any abnormalities. ARMs, tail deformities (less than half of normal length), and neural tube defects were found in 97.98% (97/99), 92.93% (92/99), and 32.32% (32/99) of the rats in the model group, respectively. The incidence of ARMs and tail deformities were 48.57% (51/105) and 35.24% (37/105), respectively, in the rescue rats; no neural tube defects were observed in these rats. The weight, body length, and tail length of the fetal rats in the rescue group were more than those in the model group, but less than those in the control group. Taken together, FA rescued ARMs induced by ATRA in rats. Thus, FA may reduce the incidence of associated malformations and improve the growth and development of fetal rats.

THE EFFECT OF LIMAN LEAVES EXTRACT ON FETAL RAT DEVELOPMENT

Objective: The objective of the study was to observe the effect of liman leaves extract on fetal rat development and provide safety information for its use during pregnancy.&#x0D; Methods: Estrus cycles of female Wistar rats were observed and mated to male rats, 0 day of pregnancy determined after finding a vaginal plug or in a vaginal smear, there was sperm. Sample was administered orally using sonde with a single administration on the 11th day of pregnancy, at doses of 3750 mg/kg bw, 1185 mg/kg bw, and 375 mg/kg bw. On the 19th day of pregnancy, the rats were sacrificed and then observed the number of implantation, corpus luteum, intrauterine death, and fetal abnormalities.&#x0D; Results: The dose of 375 mg/kg bw exposed the highest average implantation rate (13.20%) and the largest number of corpus luteum (13.90%). The highest total intrauterine death was presented by dose of 3750 mg/kg bw and significantly different (p&lt;0.05) compared to the control group. The dose of 375 mg/kg bw expressed the highest percentage of embryos with resorption (19.03%), while the lowest average fetal body was shown by 3750 mg/kg bw compared to the other groups. The highest percentage of external abnormalities was given by the dose of 375 mg/kg bw (12.91%), which abnormalities found were dwarf, cleft palate, hydrocephalus, short sleeve, and hematoma.&#x0D; Conclusion: Liman leaves extract was mild teratogenic on Wistar rat fetus.

Attempts to obtain fully xenogeneic fetuses in rat ↔ mouse model†,‡

The full-term development of the xenogeneic embryo in the uterus of the mother of different species is very restricted and can occur only in certain groups of closely related mammals. In the case of mouse ↔ rat chimeras, the interspecific uterine barrier is less hostile to interspecific chimeric fetuses. In current work, we tested the development of mouse and rat fetuses in uteri of females of the opposite species. We created chimeric mouse ↔ rat blastocysts by injection of mouse embryonic stem cells (ESCs) into eight-cell rat embryos and rat ESCs into eight-cell mouse embryos. Chimeras were transferred to the foster mothers of the opposite species. Despite a huge number of transferred embryos (>1000 in total for both variants), only one live fetus derived solely from the mouse ESCs was isolated at E13.5 from the rat uterus. All other fetuses and newborns were chimeric or were built only from the cells of the recipient embryo. We examined the possible reason for such an outcome and found that the xenogeneic fetuses are eliminated at the perigastrulation stage of development. Thus, we conclude that in the rat ↔ mouse combination even when extraembryonic tissues of the chimeric embryo are composed solely of the cells of the same species as the female to which embryos are transferred, the full-term development of the pure xenogeneic fetus is very unlikely.

Differential expression of circRNAs during rat lung development.

At present, thousands of circular RNAs (circRNAs) have been found in cancer and various tissues from different species. However, the expression of circRNAs during rat lung development remains largely unknown. In the present study, circRNA expression profiles were screened in three mixed rat lung tissues at 3 time-points [embryonic day (E) 19, E21 and post-natal (P) day 3] during fetal rat development with circRNA high-throughput sequencing. Preliminary results were verified by reverse transcription-PCR (RT-PCR) at 4 time-points (E16, E19, E21 and P3). A total of 375 circRNAs were differently expressed in E19 vs. E21 (fold change ≥1.5; P<0.05). At the same time, a total of 358 circRNAs were differently expressed in E21 vs. P3 (fold change ≥1.5; P<0.05). A total of 3 circRNAs (rno_circ:chr7:24777879-24784993, r n o _c i r c:c h r14:14 62 0 910 −14 62 49 33 a n d r n o _circ:chr3:1988750- 1998592) were characterized by having consistent fold changes (≥1.5) between 3 time-points (E19, E21 and P3) and were selected for RT-PCR at 4 time-points (E16, E19, E21 and P3). Subsequently, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis of parent genes of the differentially expressed circRNAs revealed that these circRNAs may serve important roles in lung development. The present results support that these new found circRNAs participate in lung development. Furthermore, these findings may help to clarify the physiopathological mechanism of normal rat lung development, and may further provide a physiopatho-logical basis of lung developmental diseases.

A multi-institutional study benchmarking the zebrafish developmental assay for prediction of embryotoxic plasma concentrations from rat embryo–fetal development studies

Predicting embryotoxicity of pharmaceutical compounds or industrial chemicals is crucial for public safety. Conventional studies which monitor embryo–fetal development in rats and rabbits are costly and time consuming. Alternative assays which are simpler and less costly are being pursued. The purpose of this research was to assess the capacity for the zebrafish development assay to predict mammalian plasma levels that are embryotoxic. Previously published data on rat plasma levels associated with embryotoxicity were used to guide concentration ranges for each of 25 chemicals dissolved in the media bathing developing zebrafish embryos. Embryotoxic media concentrations were compared to embryotoxic rat plasma concentrations. Assays were conducted in parallel at multiple sites as a consortium effort through the Health and Environmental Sciences Institute (HESI). Considering results from all sites, the zebrafish embryo development assay predicted (within 1-log) the rat maternal exposure levels associated with embryotoxicity 75% of the time.

The effect of Cynara scolymus (artichoke) on maternal reproductive outcomes and fetal development in rats

Cynara scolymus (C.scolymus) is a plant employed worldwide as an herbal medicine. However, there is a paucity of data related to the evaluation of its toxicity in commercial preparations; thus, the aim of this study was to evaluate the possible teratogenic effect of the dry extract of C.scolymus leaves in Wistar rats. Females were treated, from gestation day (GD) 6 until GD19, with 0.0, 1.0, 2.0 or 4.0 g/kg body weight of C.scolymus extract. At GD20, a cesarean section was performed for evaluation of maternal and fetal parameters. C.scolymus did not induce changes in food consumption, preimplantation or postimplantation losses, placental weight or biochemical profile. An increase in water consumption was observed in pregnant females treated with the higher doses of C.scolymus. Experimental groups showed lower body weight gain during pregnancy and lower gravid uterus weight. Maternal body weight minus the gravid uterus weight did not result in significant differences. Reductions in fetal weight and length were observed in experimental groups. The number of live pups per litter was lower in the highest dose group. No fetal skeletal or visceral malformations were detected. The results showed that the consumption of artichoke during pregnancy clearly has a negative impact on fetuses.

In utero Exposure to Atrazine Disrupts Rat Fetal Testis Development.

Atrazine (ATR) is a commonly used agricultural herbicide and a potential endocrine disruptor that may cause testicular dysgenesis. The objective of the present study was to investigate the effects of atrazine on fetal testis development after in utero exposure. Female Sprague-Dawley rats were gavaged daily with vehicle (corn oil, control) or atrazine (25, 50, and 100 mg/kg body weight/day) from gestational day 12 to 21. Atrazine dose-dependently decreased serum testosterone levels of male pups, with a significant difference from the control recorded at a dose of 100 mg/kg. In addition, atrazine significantly increased fetal Leydig cell aggregation at a dose of 100 mg/kg. Atrazine increased fetal Leydig cell number but not Sertoli cell number. However, atrazine down-regulated Scarb1 and Cyp17a1 in the fetal Leydig cell per se and Hsd17b3 and Dhh in the Sertoli cell per se. These results demonstrated that in utero exposure to atrazine disrupted rat fetal testis development.

In utero exposure to hexavalent chromium disrupts rat fetal testis development

Hexavalent chromium (Cr6+) acts as an endocrine disruptor. Herein, we investigated effects of Cr6+ on the development of rat fetal Leydig and Sertoli cells, which support differentiation of the male reproductive tract in late gestation. Female pregnant Sprague Dawley rats were gavaged with potassium dichromate (0, 3, 6, and 12 mg/kg) from gestational days (GD) 12 to GD 21. Leydig and Sertoli cell function was evaluated by investigating serum testosterone levels, cell number and distribution, and the expression levels of Leydig and Sertoli cell genes and proteins. Cr6+ increased serum testosterone level at dose of 3 mg/kg (1.170 ± 0.121 ng/ml vs. 0.720 ± 0.082 ng/ml in the control), while lowered it at dose of 12 mg/kg (0.400 ± 0.098 ng/ml). In addition, it showed that Cr6+ dose-dependently reduced Leydig cell size and cytoplasmic size and decreased the percentage of medium fetal Leydig cell cluster at dose of 12 mg/kg. Further study demonstrated that the expression of Leydig cell (Lhcgr, Scarb1, and Hsd3b1) and Sertoli cell (Fshr, Pdgfa, and Lif) genes in the testis was upregulated at dose of 3 mg/kg while the expression of Lhcgr, Hsd17b3 and Igf1 was downregulated by Cr6+ at dose of 12 mg/kg. In conclusion, Cr6+ had biphasic effects on fetal Leydig cell development with low dose to stimulate testosterone production and high dose to inhibit it, possibly via biphasically regulating growth factor gene expression in fetal Sertoli cells.

In utero exposure to triphenyltin disrupts rat fetal testis development

Triphenyltin is an organotin that is widely used as an anti-fouling agent and may have endocrine-disrupting effects. The objective of the current study was to investigate effects of triphenyltin on the development of rat fetal testis. Female pregnant Sprague Dawley dams were gavaged daily with triphenyltin (0, 0.5, 1, and 2 mg/kg body weight/day) from gestational day 12 to day 21. Triphenyltin dose-dependently decreased serum testosterone levels (0.971 ± 0.072 and 0.972 ± 0.231 ng/ml at 1 and 2 mg/kg, respectively) from control level (2.099 ± 0.351 ng/ml). Triphenyltin at 1 and 2 mg/kg doses also induced fetal Leydig cell aggregation, decreased fetal Leydig cell size and cytoplasmic size. Triphenyltin decreased the expression levels of Lhcgr, Scarb1, Star, Cyp11a1, Cyp17a1, Insl3, Fshr, Pdgfa, and Sox9 by 0.5 mg/kg dose and above. However, triphenyltin did not affect Leydig and Sertoli cell numbers. In conclusion, the current study indicated that in utero exposure of triphenyltin disrupted fetal Leydig and Sertoli cell development.

Antioxidant role of melatonin against nicotine's teratogenic effects on embryonic bone development.

Objective(s):This study investigated the possible effects of low (3 mg/kg) and high (6 mg/kg) doses of nicotine on the skeletal development of rat fetuses by the double staining method and the protective role of melatonin (10 mg/kg) against these effects.Materials and Methods:Eighteen adult female Wistar-Albino rats were divided into six groups (n=3, each) as control, low-dose nicotine, high-dose nicotine, low-dose nicotine+melatonin, high-dose nicotine + melatonin and melatonin. While nicotine was given to the experimental groups on gestation days 1–20, nicotine and melatonin were administered together to the treatment groups. The fetuses were delivered by cesarean section on the 20th day of pregnancy. The skeletal systems of the fetuses were stained using the double staining method. The forelimbs and hindlimbs of the fetuses were firstly investigated under a stereomicroscope, and then their photos were taken. The total bone length, the length of the ossified part and the ossification rate were calculated using the ImageJ program.Results:The degree of ossification in the bones of the feet and the hands was determined. When the total bone length and the length of the ossified part were evaluated, they were significantly decreased in the nicotine groups (P<0.05), but were close to each other in the treatment and the control groups (P<0.05). Conclusion: It has been found that the use of nicotine during pregnancy delays skeletal ossification and that melatonin, a powerful antioxidant, eliminates the teratogenic effects of nicotine.

Effects of Dietary Daidzein Supplementation on Reproductive Performance, Serum Hormones, and Reproductive-Related Genes in Rats.

The aim of this study was to investigate the effect of dietary daidzein supplementation on reproductive performance in rats. A total of twenty-four female Sprague–Dawley (SD) rats were randomly allocated to two groups and fed either with a basal diet (CON) or basal diet containing 50 mg/kg daidzein (DAI) from gestation until delivery stage. The results show that daidzein supplementation significantly increased the total litter weight and the total viable newborn weight (p < 0.05). Interestingly, daidzein supplementation acutely elevated the concentrations of serum estrogen, progesterone and insulin-like growth factor-1 (p < 0.01) after the maternal rats’ delivery. The concentrations of serum immunoglobulin A (IgA) and immunoglobulin G (IgG) were also significantly higher in the DAI maternal rats than in the CON maternal rats (p < 0.05). Moreover, daidzein significantly increased the total antioxidant capacity (T-AOC) in maternal rats’ sera and in newborns (p < 0.05) and elevated the concentration of superoxide dismutase (SOD) in both the maternal rats’ sera and their ovaries (p < 0.05). Importantly, daidzein supplementation significantly elevated the expression levels of estrogen receptor β (ERβ) and NR5A2 genes in maternal rats’ ovaries (p < 0.05) and downregulated the expression level of prolactin receptor (PRLR) in newborns (p < 0.05). These results suggest that dietary daidzein supplementation improves reproductive performance and fetal development in rats, which is associated with changes in serum hormones, tissue antioxidant capacity, and expression levels of reproductive-related genes, both in maternal rats and their offspring.

Effect of azithromycin on fetal development in rats

This study was designed to evaluate the potential risks of the Azithromycin administration on fetuses. Therefore pregnant rats were received Azithromycin orally (90 and 180 mg/kg b.wt, daily) from sixth day until the fifteenth day of gestation. At 20th day of gestation female sacrificed; abortion, fetal resorptions, visceral examination, skeletal examination, Oxidative stress markers and histopathological examination for fetuses were recorded. Azithromycin caused abortion, fetal resorptions, growth retardation, hematoma and paralysis of limbs. Also caused cerebral dilatation, microcephaly, perforation of hard palate and hemorrhage around internal organs. By skeletal examination; weak and incomplete ossification were observed in bones .Biochemical studies showed that Azithromycin administration resulted in decreased glutathione reduced but increased malonaldahyde compared to control groups. Fetal kidney revealed degeneration and necrosis in the tubular lining epithelium and fibrosis in between the atrophied renal tubules. This study supports and proofs the potential risks of the azithromycin administration on fetuses. Â

Boron enhances early embryonic gene expressions and improves fetal development of rats

Boron is present as several different components in nature. Besides its industrial use, it is an essential element and is playing a very important role in the metabolism. In this study, it was aimed to determine the in vivo effects of boron on mRNA expression of HEX, NANOG, and OCT-3/4 genes in embryo and histological changes during fetal development. Therefore, totally 60 female rats were allocated into 5 equal groups. Experimental groups are as the followings; positive control (fed with standart rat diet), negative control (fed with boron free diet), low boron group (fed with boron free diet and given 0.04 μg boron/ml via gastric gavage), marginal boron group (fed with boron free diet and given 0.3 μg boron/ml via gastric gavage) and normal boron group (fed with boron free diet and given 2 μg boron/ml via gastric gavage). Experimental period was performed for 14 days. Embryos were collected after 4 days of mating and the expression and protein levels of early embryonic genes namely HEX, NANOG, and OCT-3/4 were determined by using Real-Time PCR. Also, 10–20 day embryo and fetus development were histologically determined. According to the results, mRNA expression and protein levels of early embryonic genes were increased in boron groups while decreased in boron deficient group. Histopathologically, tissue and organ developments were definitely observed in the boron groups. In conclusion, mRNA expression levels of early embryonic genes decreased in boron deficient group and boron has an important role for fetal development.

Pravastatin alleviates lipopolysaccharide-induced placental TLR4 over-activation and promotes uterine arteriole remodeling without impairing rat fetal development.

Preeclampsia is associated with over-activation of the innate immune system in the placenta, in which toll-like receptor 4 (TLR4) plays an essential part. With their potent anti-inflammatory effects, statins have been suggested as potential prevention or treatment of preeclampsia, although evidence remains inadequate. Herewith, we investigated whether pravastatin could ameliorate preeclampsia-like phenotypes in a previously established lipopolysaccharide (LPS)-induced rat preeclampsia model, through targeting the TLR4/NF-κB pathway. The results showed that pravastatin reduced the blood pressure [maximum decline on gestational day (GD) 12, (101.33±2.49) mmHg vs. (118.3±1.37) mmHg, P<0.05] and urine protein level [maximum decline on GD9, (3,726.23±1,572.86)μg vs. (1,991.03±609.37)μg, P<0.05], which were elevated following LPS administration. Pravastatin also significantly reduced the rate of fetal growth restriction in LPS-treated rats (34.10% vs. 8.99%, P<0.05). Further pathological analyses suggested a restoration of normal spiral artery remodeling in preeclampsia rats by pravastatin treatment. These effects of pravastatin were associated with decreased TLR4/NF-κB protein levels in the placenta and IL-6/MCP-1 levels in serum. Additionally, no obvious abnormalities in fetal liver, brain, and kidney were found after administration of pravastatin. These results provide supportive evidence for use of pravastatin in preventing preeclampsia.

Estrous Cycle, Fertilility and Fetal Development in Rats with Hyperbilirubinemia

Objective: Decreasing in reproductive capacity has been observed in jaundiced female rats. The animals with hyperbilirubinemia present early sexual maturation, late ovulation, reduced number of corpora lutea and precocious vaginal opening. The interaction with toxic agents leads to abnormal development of the embryos. The objective is to investigate the influence of jaundice on estrous cycle, fertility, ovaries and fetus development. Methods: 66 female rats were divided into two groups (n=33): group 1 - ligature of the biliopancreatic duct, group 2 - sham operation. These animals were paired with males. Vaginal smears were collected daily to verify pregnancy and the gestational period. The morphologic aspect of the ovaries and the corpus luteum were studied. The morphology of the fetuses were assessed. Serum bilirubins were recorded. 32 rats of the control group and 11 jaundiced rats became pregnant. Results: The 17 rats with hyperbilirubinemia that did not become pregnant presented involutive corpora lutea and modifications in their estrous cycles, remaining in pro-estrus or estrus. The pregnant rats with hyperbilirubinemia had fetuses with abnormal development. Conclusions: Fertilization occurs in jaundiced rats, but reproductive capacity is reduced, with irregular estrous cycles, involutive corpora lutea and abnormal fetal development.

Zinc protection in fetal rats for maternal mercury exposure-induced growth retardation is probably associated with S100B expression.

The study was conducted to investigate the effects of maternal mercury exposure on fetal rat development and zinc protection in mercury-exposed rats. Pregnant rats were subjected to zinc sulfate pre-feeding, mercury exposure and zinc sulfate co-feeding. The control rats were administered distilled water. On day 19, the placental weight, overall weight, size and tail length of fetal rats, mercury content and S100B level in the placenta were determined using Western blot analysis. Compared with the control, mercury exposure at 2.0 mg/kg.d significantly reduced placental weight and fetal development, resulting in reduced fetal weight, size and tail length, while zinc pre-feeding increased placental weight and other fetal developmental parameters. Compared with mercury exposure, co-feeding with zinc significantly reduced mercury-induced injury in the fetal rats. S100B and mercury content levels were significantly elevated in rats maternally exposed to methylmercury chloride, compared with the unexposed control, while co-feeding with methylmercury chloride and zinc sulfate significantly reduced S100B and mercury levels in the placenta. Maternal exposure to mercury results in increased S100B in the placenta. Zinc sulfate feeding could reduce S100B and mercury levels, thereby protecting the rats from mercury damage. S100B level may be used to measure the antagonism between zinc and mercury during pregnancy.

Assessment of the Protective Role of Prenatal Zinc versus Insulin Supplementation on Fetal Cardiac Damage Induced by Maternal Diabetes in Rat Using Caspase-3 and KI67 Immunohistochemical Stains.

Maternal diabetes mellitus (DM) affects early organogenesis. Metabolic disorders of DM are associated with a depleted zinc status. This study evaluated the effect of maternal DM on cardiac development of rat fetuses and protective roles of prenatal zinc versus insulin supplementation. Pregnant rats were divided into 4 groups ((I) control, (II) STZ-induced DM, (III) STZ-induced DM treated with Zn, and (IV) STZ induced DM treated with insulin), all sacrificed on GD 20. Fetal heart weight of diabetic rats showed significant decrease compared to controls (P < 0.05). H&E stained section of controls had normal appearance of the myocardium, compared to diabetics that showed myocardial disarray with characteristic degenerative changes. Sections of zinc treated group showed restored architecture of normal myofibrils with minimal degenerative changes, while those of insulin treated group show partial restoration of the normal architecture of cardiomyocytes with focal improvement of cardiac tissue. Caspase-3 immunostained slides showed positive cytoplasmic immunoreactivity in diabetic group. But KI67 immunostained slides revealed negative nuclear immunoreaction in diabetics. We observed that gestational diabetes was associated with increased risk of fetal myocardial damage that might be caused by increased apoptotic level. Treating diabetic pregnant subjects with zinc and insulin was associated with improvement in myocardial integrity.