Effect of maternal diabetes and hypercholesterolemia on fetal liver of albino Wistar rats.

OBJECTIVE: The aim of this study was to predict the development of hepatic lesions and impairment of function during the development of fetuses (13-, 15-, 17-, and 19-d-old embryos) of diabetic and hypercholesterolemic mothers. METHODS: Eighty virgin and fertile male rats (one male/three females) of Wistar strain with an average body weight of 150 to 180 g were used. Mating was carried out, and pregnancy was determined by examining sperm in vaginal smears. Pregnant rats were arranged into three groups; control, diabetic (single intraperitoneal injection [i.p.] of 60 mg streptozotocin/kg) and hypercholesterolemic groups (fed on a diet containing 3% cholesterol for 6 wk before conception and throughout gestation) (n = 20). Pregnant rats were sacrificed and 13-, 15-, 17-, and 19-d-old embryos and livers were incised and subjected to histological and transmission electronic microscopical (TEM) investigations, assessments of alkaline phosphatase (Al-Pase) isoenzymes electrophoresis, DNA fragmentation, and comet assay. Flow cytometric analysis of apoptosis and caspases 3 and 9 in the livers of mother rats and their 19-d-old fetuses was determined. RESULTS: Histologic findings of diabetic and hypercholesterolemic mothers revealed apparent damage of hepatocytes, accumulation of lipid-laden cells, and vascular steatosis, while the 13-, 15-, 17- or 19-d-old fetuses of either diabetic or hypercholesterolemic mothers revealed disorganized hepatic architecture and massive cell damage. TEM of diseased mothers and their fetuses possessed increased incidence of pyknotic hepatocytes with massive vesicuolation of rough endoplasmic reticulum and degeneration of mitochondria. Al-Pase isoenzymes were altered and genomic DNA of both double and single helical structures were markedly damaged, especially in fetuses of maternally diabetic and hypercholesterolemic mothers. Flow cytometry revealed an increase in apoptosis and caspases 3 and 9 in diabetic and hypercholesterolemic mothers and their 19-d-old fetuses. CONCLUSION: These results suggested that maternal diabetes and hypercholesterolemia predicted early hepatitis and increased apoptosis in mothers and their fetuses as a result of oxidative stress and elevated apoptic markers caspases 3 and 9.

Cardiomyopathy and angiogenesis defects of Wistar rat fetuses of diabetic and hypercholesterolemic mothers.

OBJECTIVE: We aimed to illustrate the histogenesis, lactic dehydrogenase isoenzymes electrophoresis, and DNA damage of cardiac muscles and blood vessels during prenatal life of maternal diabetic or hypercholesterolemic mother. METHODS: Eighty fertile male and virgin female Wistar rats (1 male/3 females), weighing approximately 130 g, were mated and zero date of gestation was determined. Diabetes was induced at the fifth day of gestation by intraperitoneal injection of a single dose of 60 mg streptozotocin/kg body weight in citrate buffer, pH 4.6. At the same time, hypercholesterolemia was carried out by feeding virgin rats a diet containing 3% cholesterol for 6 wk before the onset of conception. Pregnant rats were arranged into three groups: control, diabetic, and hypercholesterolemic (n = 20). The animals were sacrificed and embryos were separated at 7-, 13-, 15-, 17-, and 19 d old, respectively, and subjected to light and transmission electron microscopy, lactic dehydrogenases isoenzymes electrophoresis, DNA fragmentation, and comet assay. The sera of the mothers were examined for fasting glucose level, total cholesterol, triglycerides, low-density lipoprotein, high-density lipoprotein, and creatine phosphokinase levels. RESULTS: Diabetic and hypercholesterolemic mothers exhibited a significant increase of sera cholesterol level, low-density lipoprotein, and creatine phosphokinase activity. Histologic findings of embryos of diabetic and hypercholesterolemic mothers revealed cardiomyopathy and malformation of blood vessels with an apparent degeneration of their endothelium. Transmission electron microscopy possessed massive necrosis of muscle fibers, disorganization of Z and I bands, and mitochondrial damage. Lactic dehydrogenase isoenzyme electrophoresis was altered and genomic DNA fragmentation was markedly increased. CONCLUSION: Maternal diabetes or hypercholesterolemia led to marked alterations in blood vessel differentiation as well as to cardiomyopathy during prenatal growth as assessed by the disruption of fine structures, abnormal lactic dehydrogenase isoenzymes electrophoresis, and an increase of DNA damage. These may be attributed to the marked oxidative stress and liberation of free oxygen radicals, which interrupted the myocardium structure and function during organogenesis.

Protective effects of Allium sativum against defects of hypercholesterolemia on pregnant rats and their offspring.

Sixty fertile female and male albino rats of Wistar strain (I male/ 3 females) were used in the present study. The females were divided into four groups of ten rats each. Group 1 received water and standard feeds for thirty-four days. Group 2 was fed with a cholesterol-containing diet (1%) for two weeks prior to onset of gestation and maintained administration till parturition, produce atherosclerosis (34 days). Group 3 received intragastric administration of 100mg homogenate of garlic (Allium sativum)/kg body weight for three weeks prior to onset of gestation as well as throughout the gestation period. Group 4 intragastrically administered garlic for one week of group B and maintained with combined garlic-treatment for the mentioned period. At parturition, the pregnant were sacrificed and serum total cholesterol (TCL), triglycerides (TG), HDL, LDL and creatine kinase activity (CK) were determined. The total numbers of offspring were recorded and examined morphological for congenital abnormalities. Biopsies of heart and dorsal aorta of both pregnant and their offspring (1 day-age) were processed for investigation at light and transmission electron microscopy. The skeleton of the newborn of different experimental groups were stained with alizarin red s and mor-phometric assessment of mandibular and appendicular bone length. The study revealed that the myocardium of atherosclerotic mother exhibited leuhkocytic inflammatory cell infiltration associated with necrosis, eosinophilia of myocardiai fibers, and edema of blood vessels. Ultrastructural studies revealed swelling of mitochondria, disruption of cristae in the myocardiai muscle fibers. The dorsal aorta possessed accumulation of extra-cellular lipid in intima lining of endothelium. The collagenous fibrils in the tunica adventitia became fragile and loosely separated from each other. Numerous foamy lipid loaden cells were detected within the tunica intima causing deterioration of the elastic fibers, resulting in fibrinoid necrosis. Oral supplementation with Allium sativum (100 mg/ kg) ameliorated these effects in myocardium muscle of mothers and offspring; however the dorsal aorta of mothers showed partial amelioration. Hypercholesterolemic mothers exhibited marked alterations in serum TCL, TG, LDL and CK activity. Supplementation with Allium sativum ameliorated the drastic biochemical alterations. Concerning pregnancy, hypercholesterolemia increased the incidence of abortion and abnormalities of the newborn including decreased body weight, reduced ossification of axial (mandible) and appendicular bones. All these effects were markedly ameliorated by supplementation with Allium sativum. The author finally concluded that hypercholesterolemia exhibits pathological alterations of myocardiai muscles reducing its optimal capacity for pumping blood to different body organs along with atherosclerosis of dorsal aorta which intern affect the progress of gestation and development of both morphological and skeletal abnormalities. Allium sativum-supplementation leads to amelioration of both mother and their offspring investigated parameters as a result of its antioxidant activity.