20th Day Of Gestation Research Articles

Teratogenic Effect of Crude Ethanolic Root Bark and Leaf Extracts of Rauwolfia vomitoria (Apocynaceae) on the Liver of Albino Wistar Rat Fetuses

Objectives: Rauwolfia vomitoria is a medicinal plant which is used locally for the treatment of a variety of disorders including snake and insect bites and stings, insomnia and insanity. This study was to investigate the teratogenic effect of the ethanolic root bark extract of R. vomitoria on the fetal liver. Material and methods: Twenty-five virgin female Wistar rats weighing between 180 – 200g were used. The animals were divided into 5 groups, labeled A, B, C, D and E, each consisting of 5 rats. Group A rats were the control, while Groups B, C, D and E were the experimental. The female rats at proestrous were mated with sexually matured male rats. The day after was designated as day zero of pregnancy. Groups B and C animals received oral doses of 150mg/kg and 250mg/kg body weight of the root bark extract while groups D and E animals received 150mg/kg and 250mg/kg body weight of the leaf extract respectively from 7th to 11th day of gestation. On the 20th day of gestation, the rats were sacrificed and the fetuses examined for gross anomalies. Each liver was routinely processed using Haematoxylin and Eosin method. Results: Histological observations of the fetal liver showed marked distortion of normal liver architecture in the treated groups when compared with the control, the effect being more pronounced in the groups that received the root bark extract. Conclusions: Our result suggests that ethanolic root bark and leaf extracts of Rauwolfia vomitoria maybe hepatotoxic to the developing rats liver. DOI: http://dx.doi.org/10.3126/ajms.v4i1.6848 Asian Journal of Medical Sciences 4(2013) 30-34

English

  Cadmium is a potent teratogen in laboratory animals, causing exencephaly when administered at early stages of development. Due to its heterogenicity with respect to molecular targets, the mechanisms behind cadmium toxicity are not well understood. In the present study, 40 pregnant rats (Sprague-Dawley) were divided into four groups (10 each); first group served as the control (G1), the second group (G2) received 61.3 mg/kg cadmium chloride daily from 7th to 16th day of gestation (organogenesis period) by oral tube. Group 3 (G3) was administrated a solution of 25 mg/kg zinc chloride orally from the 1st day to 20th day of pregnancy. Group 4 were administrated a solution of cadmium chloride (61.3 mg/kg) and zinc chloride (25 mg /kg) daily from the 7th to16th day of gestation. Maternal body weights were measured on gestational day 0, 6, 9, 12, 15 and 20. At the 20th day of gestation, blood samples were collected from the eye, using orbital sinus technique. Serum aspartate transaminase (AST) and alanine transaminase (ALT) were determined calorimetrically and serum, urea and creatinine were determined. All of the pregnant rats were sacrificed by ether anaesthesia at the 20th day of gestation and foetuses were removed from the uterus. The implantation sites, corpora lutea, living, dead and reabsorbed foetuses were counted and recorded. Liver of pregnant rats and their fetuses were used to isolate a total RNA for quantification of Msx1, Cx43, Bcl2 and Bax genes. The results show the toxic effect of Cd on the pregnant rats and their fetuses, at morphological, physiological and molecular level but, zinc has a very effective protection against cadmium-induced developmental toxicity.   Key words: Cadmium, zinc, rat, organogenesis, gene expression.  

Intra-amniotic LPS amplifies hyperoxia-induced airway hyperreactivity in neonatal rats.

Background:We previously showed that intra-amniotic lipopolysaccharide (LPS) amplifies alveolar hypoplasia induced by postnatal hyperoxia. We determined whether the priming effect of intra-amniotic LPS amplifies hyperoxia-induced airway hyperreactivity (AHR).Methods:LPS or normal saline was injected into the amniotic cavities of pregnant rats at the 20th day of gestation. After birth, rat pups were exposed to 60% O2 or air for 14 d. On postnatal day 14, rat pups underwent forced oscillometry, which included a challenge with nebulized methacholine, and the lungs were harvested for morphological studies.Results:Hyperoxia significantly increased airway reactivity and decreased compliance. Intra-amniotic LPS further increased hyperoxia-induced AHR but did not further impair respiratory system compliance. Hyperoxia-induced changes in lung parenchymal and small airway morphology were not further altered by intra-amniotic LPS. However, combined exposure to intra-amniotic LPS and hyperoxia increased the proportion of degranulating mast cells in the hilar airways.Conclusion:Intra-amniotic LPS amplified postnatal hyperoxia-induced AHR. This was associated with increased airway mast cell degranulation, which has previously been linked with hyperoxia-induced AHR. There were no morphologic changes of parenchyma or airways that would account for the LPS augmentation of hyperoxia-induced AHR.

Antenatal taurine supplementation reduces cerebral cell apoptosis in fetal rats with growth restriction

Objective To explore the effect of antenatal taurine supplementation on cerebral apoptosis and the expression of glial cell line-derived neurotrophic factor (GDNF) and caspase-3 in fetal rats with fetal growth restriction (FGR).Methods Fifteen pregnant Sprague-Dawley rats were randomly divided into three groups:control group,FGR model group (model group) and FGR with antenatal taurine supplementation group (taurine group).Taurine was added into the diet of taurine group at a dose of 300 mg/(kg · d) from the 12th day of gestation until natural delivery.Two appropriate for gestational age (AGA) newborn rats were randomly selected from each mother in control group and two FGR fetal rats were randomly selected from each mother both in model and taurine groups.Apoptosis of neural cells in the brain was detected by terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL).Changes in protein expression of GDNF and caspase-3 were detected by immunohistochemistry.Levene method,one-way ANOVA and SNK-test,or Kruskal Wallis rank sum test and Tamhane’ s test were applied for statistical analysis.Results (1)The total amounts of fetal rats in control group,model group and taurine group were 65,60 and 59.The mean body weight of fetal rats were (6.36±0.44) g,(4.55 ± 0.45) g and (5.11±0.67) g,respectively.All fetal rats developed FGR in model group,while 76.3％(45/59) of fetal rats were FGR in taurine group.Therefore,FGR model was successfully established.(2) In control group,there were few expression of TUNEL positive cells in cerebral cortex.A large amount of TUNEL positive cells were found in the cortex,hippocampal and white matter area in model group,but less positive cells were identified in taurine group than in model group.The amount of apoptotic brain cells in the three groups were (0.46 ± 0.11),(14.76 ± 3.42) and (6.78 ± 1.93),respectively(H=429.80,P=0.000).(3)There were only small amount of GDNF positive cells in cerebral cortex in control group and more in model group.The amount of GDNF positive cells was further increased in taurine group.The amount of GDNF positive cells in cerebral cortex in the three groups were (93.56± 6.73),(120.36± 6.23)and(139.56± 5.28),respectively (H=715.17,P=0.000).(4) Few caspase-3 positive cells were found in cerebral cortex in control group.A large amount of positive cells were found in model group and less positive cells were found in taurine group,which was still more than those in control group.The amounts of caspase-3 positive cells in the three groups were (7.50±2.31),(151.32±24.43)and(37.28±11.21),respectively (F=132.54,P=0.000).Conclusions The number of apoptotic neural cells in brain tissue of baby rats with FGR were significantly increased,which can be significantly alleviated by maternal antenatal taurine supplementation through upregulation of GDNF and downregulation of caspase-3 expression. Key words: Fetal growth retardation; Taurine; Apoptosis; Glial cell line-derived neurotrophic factor; Caspase 3

Expression of transient receptor potential channels in the ependymal cells of the developing rat brain

Cerebrospinal fluid (CSF) plays an important role in providing brain tissue with a stable internal environment as well as in absorbing mechanical and thermal stresses. From its initial composition, derived from the amniotic fluid trapped by the closure of neuropores, CSF is modified by developing and differentiating ependymal cells lining the ventricular surface or forming the choroid plexus. Its osmolarity and ionic composition brings about a change through the action of many channels expressed on the ependymal cells. Some newly discovered transient receptor potential (TRP) channels are known to be expressed in the choroid plexus ependyma. To detect additional TRP channel expression, immunohistochemical screening was performed at the choroid plexus of 13-, 15-, 17-, and 19-day embryos, using antibodies against TRPV1, TRPV3, and TRPA1, and the expression was compared with those in the adult TRP channels. The level of TRP channel expression was higher in the choroid plexus which suggests more active functioning of TRP channels in the developing choroid plexus than the ventricular lining ependyma in the 15- and 17-day embryos. All the expression of TRP channels decreased at the 19th day of gestation. TRPA1 was expressed at a higher level than TRPV1 and TRPV3 in almost all stages in both the choroid plexus and ventricular lining epithelium. The highest level of TRPV1 and TRPV3 expression was observed in association with the glycogen deposits in the cytoplasm of the choroid plexus ependymal cells of the 15- and 17-day embryos.

Effect of allograft amniotic membrane use on adhesion formation after Cesarean section in pregnant rat

We aimed to investigate the effect of allograft amniotic membrane utilization to prevent the post-operative adhesion formation. In 24 pregnant inbreed Wistar-Albino rats, pregnancy was terminated by forming bilateral uterine horn defect via cesarean section at 20th gestational day. Rats were assigned in three groups randomly. In the first group, abdomen closure was achieved without administration of any intra-peritoneal material following standard surgical intervention. In the second group Seprafilm was used to cover the defect at anterior horn of uterus; whereas amniotic membrane of the rat itself was used in the third group. After 3 weeks, all rats were sacrificed and re-laparotomy was performed to determine adhesions scores. No significant difference was found in adhesion scores between the group 1 and group 3 and also between group 2 and group 3, supporting the previous findings in the literature. We observed that direct application of allograft amniotic membrane, which is an adhesion barrier used after cesarean section, to injured surface had no effect in the prevention of adhesions (Tab. 3, Fig. 5, Ref. 28).

Hepatic damage in newborns from female rats exposed to the pesticide derivative ethylenethiourea

To evaluate hepatic morphological-histological abnormalities in newborns from female rats exposed to ethylenethiourea. A randomized study was conducted on fifty-five newborn Wistar rats were studied: 34 in the experimental group, whose mothers had been exposed to 1% ethylenethiourea; and 21 in the control group, whose mothers had received 0.9% physiological solution. The solution was administered via gavage on the 11(th) day of gestation. Cesarean section was performed on the 20(th) day of gestation. The newborns' livers were examined and any morphological-histological abnormalities were registered. The presence of megakaryocytes was quantified in 50 microscope fields, as the total number of these cells per mm(2). The entire experimental group presented abnormalities of embryonic formation, with musculoskeletal anomalies, digestive system anomalies, hepatic congestion and friability, hydrops and delayed intrauterine growth. The histopathological analysis showed that morphological-histological hepatic destructuring had occurred in all entire experimental with removal of the hepatic trabeculae and severe hepatic megakaryocytosis. The mean megakaryocyte density ranged from 107.9 to 114.2 per mm(2), and it was eight times greater than in the control group, thus characterizing a situation of extramedullary hematopoiesis. The fetal exposure to ethylenethiourea caused hepatic damage characterized by severe extramedullary hematopoiesis.

Distribution of Zonula Occludens‐1 and Occludin and alterations of testicular morphology after in utero radiation and postnatal hyperthermia in rats

In utero irradiation (IR) and postnatal hyperthermia (HT) exposure cause infertility by decreasing spermatogenic colony growth and the number of sperm in rats. Four groups were used: (i) Control group, (ii) HT group (rats exposed to hyperthermia on the 10th postnatal day), (iii) IR group (rats exposed to IR on the 17th gestational day) and (iv) IR+HT group. Three and six months after the procedures testes were examined by light and electron microscopy. Some degenerated tubules in the HT group, many vacuoles in spermatogenic cells and degenerated tight junctions in the IR group, atrophic tubules and severe degeneration of tight junctions in the IR+HT group were observed. ZO-1 and occludin immunoreactivity were decreased and disorganized in the HT and IR groups and absent in the IR+HT group. The increase in the number of apoptotic cells was accompanied by a time-dependent decrease in haploid, diploid and tetraploid cells in all groups. Degenerative findings were severe after 6months in all groups. The double-hit model may represent a Sertoli cell only model of infertility due to a decrease in spermatogenic cell and alterated blood-testis barrier proteins in rat.

Protection of rat fetuses by quercetin against caffeine- induced cleft palate

Caffeine at high doses is a known rodent teratogen and it induces limb malformations along with cleft palate in various strains of rats and mice. The teratogenic effects of some drugs can be prevented by the application of antioxidant drugs and stimulation of the maternal immune system. Also, there are some evidence that quercetin is an antioxidant. Therefore, in this study, the prophylactic effect of quercetin on teratogenic effects of caffeine was evaluated. This study was performed on 26 pregnant rats that were divided into four groups. Control group received normal saline and test groups received caffeine (80 mg/kg), caffeine (80 mg/kg) plus quercetin (75 mg/kg), and quercetin (10 mg/kg), intraperitoneally at 9 to 11th days of gestation, respectively. Fetuses were collected at the 20th day of gestation and after determination of weight and length; they were stained by Alizarin red - Alcian blue method. Cleft palate incidence was 33.33% in fetuses of rats that received only caffeine and incidence was 1.49% in the group that received caffeine plus quercetin (75 mg/kg). The means of weight and length of fetuses from rat that received quercetin were significantly greater than those that received only caffeine. It is concluded that quercetin decreased cleft palate induced by caffeine, but this subject needs more detailed evaluation. Key words: Caffeine, quercetin, pregnancy, cleft palate, teratogenicity, fetus, rat.

The Antioxidant Capacity and Anti-diabetic Effect of Boswellia serrata Triana and Planch Aqueous Extract in Fertile Female Diabetic Rats and the Possible Effects on Reproduction and Histological Changes in the Liver and Kidneys

Boswellia serrata has been used in a wide variety of diseases, including diabetes mellitus and inflammatory diseases. This study focused on the effects of Boswellia serrata aqueous extract on blood glucose and the complications of diabetes in the liver and kidneys and examined the impact of plant on reproduction in diabetic rats. The antioxidant capacity of plant extract was performed using FRAP assay. Diabetic and control rats were administered 200, 400, and 600 mg/kg Boswellia serrata extract. Vaginal plaque was mentioned as a positive sign of pregnancy ;and treatment started with extract or vehicle from 1th to 17th day of gestation by gastric gavage. Blood glucose was measured during 17 days. The Administration of Boswellia serrata in diabetic rats significantly decreased the level of blood glucose and HbA1c after 17th days (P ≤ 0.01). In diabetic group that received no treatment, the abortion of fetus spontaneous was 19.14%. The percentage of absorptions significantly was elevated in vehicle-treated diabetic rats, in comparison with vehicle- treated healthy rats. In the diabetic group, separated necrosis of hepatocytes, anarchism of liver plates, and lymphocytic inflammation were improved. Diabetic complications were not seen and the severity of damage was reduced. These damages include: lymphocytic inflammation in the port areas, irregularities, apoptosis of liver cells, and dilatation of the sinusoids. The results suggest that Boswellia serrata extract has the antidiabetic effects and can prevent the complications of diabetes in the kidneys and liver.

402 Effect of Antenatal Corticosteroids in Activity and Expression of Secretory Phospholipase A2 and TNF Alfa in Lung of Newborn Rats

IntroductionThe sPLA2 plays an important role in the development of acute respiratory distress syndrome. It is regulated by many factors including steroids and TNFa. Antenatal corticosteroids are recommended for preventing...

The effects of caffeine and carvedilol on skeletal system of rat embryos in prenatal period

Caffeine at high doses is a known rodent teratogen and induces limb malformations along with cleft palate in various strains of rats and mice. The teratogenic effects of some drugs can be prevented by the application of antioxidant drugs and stimulation of the maternal immune system. Also, there are some evidence that carvedilol is antioxidant. Therefore, in this study, the prophylactic effect of carvedilol on teratogenic effects of caffeine was evaluated. This study was performed on 24 pregnant rats that were divided into four groups. Control group received normal saline and test groups received caffeine (80 mg/kg), caffeine (80 mg/kg) plus carvedilol (5 mg/kg) and carvedilol (5 mg/kg), intraperitoneally at 9 to 11th days of gestation, respectively. Fetuses were collected at 20th day of gestation and after determination of weight and length, they were stained by Alizarin red - Alcian blue method. Cleft palate incidence was 33.33% in fetuses of rats that received only caffeine, while it was 2.85% in group which received caffeine plus carvedilol (5 mg/kg). The means of weight and length of fetuses from rats that received carvedilol were significantly greater than those that received only caffeine. It is concluded that carvedilol decreased cleft palate induced by caffeine, but this subject needs more detailed evaluation. Key words: Caffeine, carvedilol, gestation, cleft palate, teratogenicity, fetus, rat.

PP074. Characteristic changes in endothelial function in uterine artery by administration of no synthase inhibitor during period of placental formation

Preeclampsia is characterized as an increased peripheral vascular resistance. It is found that the administration of the NOS inhibitor l-NAME at early pregnancy developed hypertension in rats similar to preeclampsia. Furthermore, we found action of endothelial NO was reduced due to possibly reactive oxygen species (ROS) in preeclamptic women. We investigated whether this treatment modulates NO and ROS production in uterine artery endothelial cells at late pregnancy in rats. l-NAME was continuously administered during the 8-14th day of gestation in Wistar rats. Blood pressure was meassured by use of tail-cuff method. The uterine artery was obtained at the 20th day of gestation. Acetylcholine (ACh)-induced endothelial NO production was estimated from DAF-2 fluorescence. Furthermore, ACh-induced endothelial cell ROS production was estimated from CM-H2 DCFDA by fluorescence imaging system. Blood pressure was higher and birth body weight was smaller in l-NAME-treated rats than in saline-treated rats. Acetylcholine (ACh)-induced endothelial NO production was higher, while ACh-induced endothelial cell ROS production was similar, in l-NAME-treated rats when compared with those in control rats. It is suggested that the increased endothelial NO production in the uterine artery seen by a short term administration of l-NAME to rats during early pregnancy may function to protect the development of hypertension.

Efeito da administração pré-natal da dexametasona em ratas sobre os perfis glicídicos e hematológicos materno e da prole

Avaliou-se o efeito da administração de dexametasona no início e no meio da gestação de ratas, sobre os perfis glicídicos e hematológicos materno e da prole. Os animais foram submetidos aos seguintes tratamentos: dexametasona do primeiro ao sétimo dia e placebo do oitavo ao 14º dia; placebo do primeiro ao sétimo dia e dexametasona do oitavo ao 14º dia; dexametasona do primeiro ao 14º dia e placebo do primeiro ao 14º dia de gestação. A dexametasona foi administrada por via intraperitoneal, na dose de 0,8mg/kg. Foram coletadas amostras de sangue no sétimo, 14º e 21º dias de gestação, e de sangue e tecido hepático da prole no quinto, 10º e 15º dias pós-natal. Para a verificação das reservas de glicogênio hepático da prole, cortes histológicos foram corados pelo ácido periódico de Schiff. Os resultados apontam para um efeito tempo-dependente da administração de dexametasona durante a gestação, levando a alterações temporais distintas na hematologia e na concentração plasmática de carboidratos nas matrizes e na prole.

The protective effect of Chrysanthemum fantanesii extract, vitamin E and C on sodium valproate-induced embryotoxicity in pregnant mice

Valproate (VPA) has been shown to induce neural tube defect in human and mice. In this study, we investigated the ability of butanolic extract from leaves of Chrysanthemum fantanesii, vitamin E and C to modulate VPA-induced toxicity and oxidation damage in maternal and fetal tissues in mice. Plant extract, VPA and vitamins were administered orally to pregnant mice from the 6 to 17th day of gestation. Animals received plant extract (200 mg/kg per day), vitamin E (100 mg/kg per day) and vitamin C (8.3 mg/kg per day) with an oral administration of VPA (400 mg /kg per day) under the same conditions. On day 18 of gestation pregnant mice were sacrificed, fetuses, placenta and maternal tissues were removed, homogenized and used for the determination of lipid peroxidation rates (LPO) using thiobarbituric acid reactive substance (TBARS) method. Embryotoxicity wasassessed by counting the number of implants, live and dead fetuses, and resumptions. Thefetuses were observed for malformation including neural tube defect (Exencephaly), open eyes and skeletal malformation. The results show clearly that there is a positive correlation between the increase in LPO and congenital malformation. Plant extract, vitamin E and C caused partial decrease of embryo toxicity and congenital malformation induced by VPA in mice. Key words: Valproic acid, congenital malformation, embryotoxicity, oxidative stress, lipid peroxidation, Chrysanthemum fantanesii, vitamin E and C.

730 THE ROLE OF JARID1A IN SPERMATOGENESIS EVALUATION OF TRANSCRIPTIONAL REGULATION BY HISTONE H3K4 METHYLATION

You have accessJournal of UrologyPediatrics: Basic Research1 Apr 2012730 THE ROLE OF JARID1A IN SPERMATOGENESIS EVALUATION OF TRANSCRIPTIONAL REGULATION BY HISTONE H3K4 METHYLATION Hidenori Nishio, Yoshiyuki Kojima, Yoshinobu Moritoki, Makoto Imura, Hideyuki Kamisawa, Atsushi Okada, Kentaro Mizuno, Yukihiro Umemoto, Keiichi Tozawa, Yutaro Hayashi, and Kenjiro Kohri Hidenori NishioHidenori Nishio Nagoya, Japan More articles by this author , Yoshiyuki KojimaYoshiyuki Kojima Nagoya, Japan More articles by this author , Yoshinobu MoritokiYoshinobu Moritoki Nagoya, Japan More articles by this author , Makoto ImuraMakoto Imura Nagoya, Japan More articles by this author , Hideyuki KamisawaHideyuki Kamisawa Nagoya, Japan More articles by this author , Atsushi OkadaAtsushi Okada Nagoya, Japan More articles by this author , Kentaro MizunoKentaro Mizuno Nagoya, Japan More articles by this author , Yukihiro UmemotoYukihiro Umemoto Nagoya, Japan More articles by this author , Keiichi TozawaKeiichi Tozawa Nagoya, Japan More articles by this author , Yutaro HayashiYutaro Hayashi Nagoya, Japan More articles by this author , and Kenjiro KohriKenjiro Kohri Nagoya, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.815AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Spermatogenesis, a fundamental process in the male reproductive system, requires a series of tightly controlled epigenetic and genetic events in germ cells ranging from spermatogonia to spermatozoa. We recently reported that important differentiation occurs in the testes of rats at approximately 9 days. We observed through microarray gene expression analysis that Jumonji, AT rich interactive domain 1A (Jarid1a) was highly expressed in the undescended testes compared with normal testes at 9 days. The purpose of this study was to examine the effects of Jarid1a, a key epigenetic regulator expressed in the testis, on spermatogenesis. METHODS We injected flutamide (7.5 mg/day per body) into the abdomen of pregnant rats for 7 days from the 14th to 20th day of gestation to obtain offspring with undescended testes. Nine-day-old male rats were divided into 3 groups: (1) f-UDT, offspring with undescended testes, (2) f-DT, offspring with descended testes, and (3) nD9, normal testes of normal rats. Jarid1a gene expression in these 3 groups was assessed by real-time polymerase chain reaction (RT-PCR) and western blot. The localization of expression was assessed by immunohistochemistry. Moreover, these groups were assessed by western blot using the anti-histone H3 lysine 4 (H3K4) antibody and methylated antibody to evaluate the state of H3K4 modification. RESULTS RT-PCR study showed that Jarid1a was significantly expressed in the f-UDT compared to f-DT or nD9. Similar findings were observed by western blot study analysis. Immunohistochemical analysis showed that Jarid1a was expressed in the nuclei from Ad spermatogonia to spermatids. Furthermore, western blot analysis of H3K4 methylation showed that levels of dimethylated H3K4 (H3K4me2) and tri-methylated H3K4 (H3K4me3) were higher in f-UDT than in f-DT or nD9. CONCLUSIONS Jarid1a belongs to the Jumonji family, whose members share the Jumonji C (JmjC) domain and suppress transcription by means of demethylating tri- and di-methylated H3K4. Because Jarid1a was expressed in rat testes in widespread spermatogenesis and was especially high in early spermatogenesis, the state of methylation of H3K4 highly correlates with spermatogenesis. Jarid1a demethylase-regulated histone modification is likely to inhibit spermatogenesis via a low level of methylated H3K4 in undescended testes. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e300 Peer Review Report Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hidenori Nishio Nagoya, Japan More articles by this author Yoshiyuki Kojima Nagoya, Japan More articles by this author Yoshinobu Moritoki Nagoya, Japan More articles by this author Makoto Imura Nagoya, Japan More articles by this author Hideyuki Kamisawa Nagoya, Japan More articles by this author Atsushi Okada Nagoya, Japan More articles by this author Kentaro Mizuno Nagoya, Japan More articles by this author Yukihiro Umemoto Nagoya, Japan More articles by this author Keiichi Tozawa Nagoya, Japan More articles by this author Yutaro Hayashi Nagoya, Japan More articles by this author Kenjiro Kohri Nagoya, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Developmental toxicity evaluation of methanol extracts of Carthamus tinctorius L. in the Balb/C pregnant mice during organogenesis period

The plant Carthamus tinctorius L. (Safflower), a member of the compositae is used as a food additive but also has medicinal applications. To assay its effects during the developmental phase, 80 pregnant mice were randomly divided into one control and three experimental groups (n= 20). The control group received only distilled water, whereas experimental groups were administered 0.7, 1.4, and 2.8 mg/kg/day of methanol extract of C. tinctorius L. from the 6th to 16th day of gestation intraperitoneally (i.p.), respectively. On the 17th day of pregnancy, all of the animals were euthanized and the fetuses were removed and developmental study was performed. C. tinctorius caused decreased fetal body weight, crown rump and tail lengths (p < 0.05) at 1.4 and 2.8 mg/kg/day significantly compare to control group. Skeletal malformations and anomalies included hind limb flexure, exencephally and rhinencephaly were also seen. It was concluded that methanolic extract of C. tinctorius L. has toxic effects during developmental phase on the mice embryo that should be considered when used as a treatment for gynecological disease in pregnant. Key words: Carthamus tinctorius L., Methanolic extract, developmental toxicity, mice, organogenesis.

Comparison of effects of prenatal administration of dexamethasone and ambroxol on expression of Toll-like receptor 4 of fetal and neonatal rats

Objective To investigate the role of prenatal single-dose administration of dexamethasone and ambroxol on the expression of Toll-like receptor 4 (TLR4) of fetal and neonatal rats. Methods Fifty-four pregnant rats were randomly divided into three groups with eighteen rats in each group:rats treated with 0.2 mg/kg dexamethasone (group 1),0.2 mg/kg dexamethasone and 100 mg/kg ambroxol (group 2),or saline(controls) on the 17th day of gestation.The lung tissues of the offsprings were harvest independently on the 19th day of gestation,the postnatal 3 days and 7 days.The expressions of TLR4 in fetal/neonatal rat lungs of each pregnant rat were analyzed by reverse transcription-polymerase chain reaction(RT-PCR),immunohistochemistry stain,and Western blot. ANOVA and two independent samples t-test were applied. Results On the 19th day of pregnancy,TLR4 mRNA expression was up-regulated in lungs of the two treatment groups compared with controls(controls:0.26 ± 0.18,group 1:0.39 ± 0.21,t =5.866,P＜ 0.05 ; control:0.27 ± 0.22,group 2:0.46 ± 0.13,t =9.572,P＜ 0.01 ).TLR4 mRNA expression was up-regulated in group 2 compared with controls on the postnatal 3 days and 7 days(postnatal 3 d:0.59 ± 0.23 and 0.47 ±0.24,t=2.295,P＜0.05;postnatal 7 d:0.52±0.12 and 0.35±0.17,t=4.219,P＜0.05),while no significant difference was found in group 1 compared with the controls(postnatal 3 d:0.45±0.22 and 0.44±0.14,t=0.128,P＞0.05; postnatal 7 d:0.40±0.16 and 0.36 ±0.12,t=1.365,P＞0.05).Results of the immunohistochemistry demonstrated that on the 19th day of pregnancy,the protein expression of TLR4 was significantly increased in the two treatment groups (controls:0.20 ± 0.29,group 1:0.35±0.32,t=7.179,P＜0.05 ;controls:0.20±0.29,group 2:0.39±0.25,t=10.764,P＜0.01).The protein expression of TLR4 was significantly increased in group 2 on the postnatal 3 days and 7 days(postnatal 3 d:0.55±0.32 and 0.37±0.18,t=7.121,P＜0.05;postnatal 7 d:0.41±0.29and 0.25±0.24,t=6.355,P＜0.05),while no notable difference was found between group 1 and the control (postnatal 3 d:0.40±0.21 and 0.37±0.18,t=0.683,P＞0.05 ;postnatal 7 d:0.28±0.31 and 0.25±0.24,t=0.462,P＞0.05).Results of the Western blot demonstrated that on the 19th day of pregnancy,the protein expression of TLR4 was significantly increased in the two treatment groups (controls:0.15 ± 0.12,group 1:0.27± 0.20,t =7.835,P＜0.05; controls:0.16 ± 0.18,group 2:0.34±0.16,t=10.470,P＜0.01).The protein expression of TLR4 was significantly increased in lungs of the combination administration group on the postnatal 3 days and 7 days(postnatal 3 d:group 2:0.37±0.20 and 0.25±0.22,t=6.379,P＜0.05; postnatal 7 d:0.35±0.15 and 0.24±0.13,t=5.152,P＜0.05),while no notable difference could be found between group 1 and the control (postnatal3 d:0.32±0.26 and 0.25±0.16,t=1.167,P＞0.05; postnatal 7 d:0.29±0.19 and 0.24±0.10,t =1.248,P ＞ 0.05 ). Conclusions Prenatal single-dose administration of dexamethasone may up-regulate the expression of TLR4 in the rat fetal lung.The up-regulation of TLR4 might be one of the critical factors for glucocorticoid-induced maturity of fetal lung.Prenatal single-dose administration of dexamethasone and ambroxol may have effects on the regulation of TLR4 not only in fetal rats,but also in neonatal rats. Key words: Dexamethasone; Ambroxol; Lung; Toll-like receptor 4; Fetal organ maturity; Rats

The evaluation of teratogenicity of nanosilver on skeletal system and placenta of rat fetuses in prenatal period

Nanosilver, comprising silver nanoparticles, is attracting interest for a range of biomedical applications owing to its potent antibacterial activity. It has recently been demonstrated that nanosilver has useful anti-inflammatory effects and improves wound healing, which could be exploited in developing better dressings for wounds and burns. The teratogenicity potential for intraperitoneally dose of nanosilver had not previously been evaluated. Despite the widespread use of nanosilver products, relatively few studies have been undertaken to determine the biological effects of nanosilver exposure. The purpose of this study was to evaluate the teratogenicity of nanosilver in rat fetuses. This study was performed on 30 pregnant rats that were divided into five groups. Control group received normal saline and test groups received nanosilver (0.4 and 0.8 mg/kg) intraperitoneally at 8 and 9th day of gestation, respectively. Fetuses were collected at 20th day of gestation. After determination of weight and length; the fetuses were stained by Alizarin red-Alcian blue method. Also, placenta were weighed, and width and volume were measured and examined macroscopically. The mean of weight of animals' fetuses that received nanosilver (0.4 and 0.8 mg/kg) in 8th day and weight and length (0.8 mg/kg) in 9th day was significantly decreased in comparison with normal saline group. The weight, volume and width of placentas in treated animals were lesser than in control group. No macroscopic anomalies were seen in all the groups. Thus, nanosilver had no effect on skeletal system of rat fetuses and it is necessary to determine the association between the period of exposure and histopathologic changes with different doses over different time periods. Key words: Nanosilver, pregnancy, placenta, teratogenicity, rat.

Early undernutrition induces glucagon resistance and insulin hypersensitivity in the liver of suckling rats

Developing brains are vulnerable to nutritional insults. Early undernutrition alters their structure and neurochemistry, inducing long-term pathological effects whose causal pathways are not well defined. During suckling, the brain uses glucose and ketone bodies as substrates. Milk is a high-fat low-carbohydrate diet, and the liver must maintain high rates of gluconeogenesis and ketogenesis to address the needs of these substrates. Insulin and glucagon play major roles in this adaptation: throughout suckling, their blood concentrations are low and high, respectively, and the liver maintains low insulin sensitivity and increased glucagon responsiveness. We propose that disturbances in the endocrine profile and available plasma substrates along with undernutrition-related changes in brain cortex capacity for ketone utilization may cause further alterations in some brain functions. We explored this hypothesis in 10-day-old suckling rats whose mothers were severely food restricted from the 14th day of gestation. We measured the plasma/serum concentrations of glucose, ketone body, insulin and glucagon, and hepatic insulin and glucagon responses. Undernutrition led to hypoglycemia and hyperketonemia to 84% (P < 0.001) and 144% (P < 0.001) of control values, respectively. Liver responsiveness to insulin and glucagon became increased and reduced, respectively; intraperitoneal glucagon reduced liver glycogen by 90% (P < 0.01) in control and by 35% (P < 0.05) in restricted. Cortical enzymes of ketone utilization remained unchanged, but their carrier proteins were altered: monocarboxylate transporter (MCT) 1 increased: 73 ± 14, controls; 169 ± 20, undernourished (P < 0.01; densitometric units); MCT2 decreased: 103 ± 3, controls; 37 ± 4, undernourished (P < 0.001; densitometric units). All of these changes, coinciding with the brain growth spurt, may cause some harmful effects associated with early undernutrition.