Effect of prenatal administration of NSAIDs on the immune response in juvenile and adult rats

In order to investigate the effect of non-steroidal anti-inflammatory drugs (NSAIDs) on the development of rat immunity, indomethacin (IND; 0.25, 0.5, or 1.0 mg/kg/day), acetyl salicylic acid (ASA; 90, 180, or 360 mg/kg/day), or diclofenac sodium salt (DSS; 0.5, 1.0, or 2.0 mg/kg/day) suspended in 0.5% methylcellulose aqueous solution, was orally administered once daily to five pregnant Sprague-Dawley (IGS) rats per group on days 18-21 of gestation. After parturition, the serum IgM and IgG levels, the spleen weight, and the number of spleen cells were measured in 3- and 8-week-old pups. Afterwards, immunophenotyping analysis of splenocytes or peripheral blood lymphocytes and T-dependent antibody response were performed. The number of spleen cells in 3-week-olds increased when 1.0 mg/kg of IND and 180 mg/kg of ASA were administered. Immunophenotyping analysis using flow cytometry (FCM) indicated that the proportion and number of CD45RA(+) cells increased, and the proportion of CD3(-) NKR-P1A(+) cells decreased in males when dosed with IND at 1.0 mg/kg or ASA at 180 mg/kg. The serum anti-KLH IgG antibody titer decreased in the males of the IND 1.0 mg/kg dosing group, the serum levels of anti-KLH IgM, total IgM, and IgG were not changed at all. These changes disappeared in 8-week-old pups. There were no effects on any of the parameters in the 3- and 8-week-olds of the DSS treatment group. These results suggest that IND or ASA administration to dams during late gestation either causes a change in the lymphocyte subsets, or that they suppress the T-dependent antibody response in juvenile males. Both of these changes eventually recover to intact levels later on during development. These results will contribute to the development of a technique for the assessment of developmental immunotoxicity and generate data on the effect of prenatal administration of NSAIDs on the developmental immune system in pups.     

Developmental immunotoxicity of dexamethasone: comparison of fetal versus adult exposures

Dexamethasone-21 phosphate was administered (s.c.) to pregnant CD rats at days 6-21 of gestation (0, 0.0625, 0.125, 0.25, and 0.5 mg/kg/day) with identical exposure of non-pregnant adult females. Some reproductive (anogenital distance) and growth (body weight) measures of pups were altered. In the juvenile (5 weeks), the delayed type hypersensitivity response to KLH was significantly reduced at all doses examined and this pattern continued into adulthood (13 weeks). In contrast, the DTH response of adults exposed to DEX was unaltered even at the highest dose. Few DEX-induced changes were seen in offspring or adult blood parameters or in splenocytes analyzed for cell surface makers (by flow cytometry). The thymus of both exposed pups (both ages) and adults showed a marked reduction in the medulla/lobe area beginning with the 0.125 mg/kg/day DEX exposure level. Macrophage production of TNF and NO was only marginally affected as was splenocyte production of IL-4 and IFN-gamma. In contrast, pups assessed as juveniles were significantly depressed in splenic IL-2 and IL-10 production. DEX exposure altered serum antibody levels across age groups with an increase of KLH-specific IgG (beginning with the 0.0125 mg/kg/day dose) while total IgE was reduced. These results suggest that while DEX exposure produces some common alterations following in utero versus adult exposure, fetal exposure (even at the lowest doses tested) produces marked and persistent functional loss (DTH) not evident in exposed adults. Furthermore, there was no apparent advantage in delaying immune assessment until the offspring reached adulthood.     

Effect of fosfomycin-Na on reproductive performance of rats. II. Fertility test (author's transl)]

The fertility study of fosfomycin-Na (FOM-Na) was undertaken in Wistar rats. FOM-Na was administered intraperitoneally at dose levels of 125, 250, 750 and 1,500 mg/kg/day. The male rats were continuously treated with FOM-Na from 63 days before mating and the females were treated from 14 days before mating. The males and females were treated through the mating period and then the pregnant rats were treated until 7 days of gestation. All pregnant rats were sacrificed on day 20 of gestation and then their fetuses were examined for external, visceral and skeletal observation. Mean body weight of males decreased and 6 males and 3 females died in the maximum dose. However, no significant differences were found between treated groups and the control with regard to fertility rate and pregnant rate. External, visceral and skeletal anomalies related with FOM-Na treatment were not observed. In this experiment, no effects of FOM-Na for reproductive performance in rats were observed.     

Exposure to lead during critical windows of embryonic development: differential immunotoxic outcome based on stage of exposure and gender.

Previous rat studies with lead (Pb) have shown that exposure throughout the full gestational period results in persistent immunotoxicity detectable in both juvenile and adult offspring. Gender differences are also evident. However, little is known about the persistent immunotoxic effects of Pb when administered during specific stages of embryonic development. Adult Sprague-Dawley female rats were administered Pb acetate (or control acetate) in their drinking water early in gestation (days 3-9) or late in gestation (days 15-21). Significantly depressed delayed type hypersensitivity (DTH) responses as well as elevated IL-10 production, relative monocyte numbers, and increased relative thymic weights were observed in late-gestation Pb-exposed female offspring assessed as adults. In contrast, late-gestation Pb-treated male offspring had significantly increased IL-12 production and decreased IL-10 production, while the DTH response, relative monocyte numbers and thymic weights were unchanged. With early exposure, the primary alteration was decreased nitric oxide production in Pb-treated males, whereas in Pb-treated females nitrite production was unaltered. These results suggest that at the Pb dosage employed, the embryo may be more sensitive to the full range of Pb-induced immunotoxic effects with late gestational Pb exposure, and the effects of Pb on DTH function are more pronounced in females. The data also indicate that adherent splenocytes (probably macrophages) and T lymphocytes are the primary immune cells affected during fetal Pb exposure, and that gender may influence the impact of Pb exposure on these cells. Therefore, additional developmental immunotoxicity studies are needed to examine critical windows of immune development for immunotoxicity and differential susceptibility based on gender.     

Impact of high flaxseed diet on mitogen-induced proliferation, IL-2 production, cell subsets and fatty acid composition of spleen cells from pregnant and F1 generation Sprague-Dawley rats.

Flaxseed (FS) being rich in alpha-linolenic acid may alter the immune parameters. Therefore, we assessed the impact of FS and defatted flaxseed meal (FLM) on fatty acid composition, cell subsets, proliferation and IL-2 production by splenic lymphocytes. Pregnant female Sprague-Dawley rats were fed diets containing 0% FS and FLM, 20 or 40% FS, 13 or 26% FLM during gestation or gestation, lactation and 8 week post-weaning period. FS and FLM resulted in up to 8.3 fold and 4.6 fold increase in splenic ALA among pregnant rats, 4.5 fold and 1.2 fold increase in splenic ALA among F(1) generation rats. Splenic linoleic acid (LA) and arachidonic acid (AA) were 18 and 40% lower in 40% FS fed pregnant rats, and AA was 15% lower in all the other groups. Among F(1) rats, splenic LA and AA were 16 and 48% lower in 40% FS group, and AA was 18% lower in 20% FS and 26% FLM groups. Concanavalin A and phytohemagglutinin mediated proliferation of spleen cells were 60 and 52% lower in 40% FS fed pregnant and F(1) generation rats, respectively. No significant changes were observed in the cell subsets or IL-2 production by splenic cells from different groups.     

Embryotoxic evaluation of a Prudhoe Bay crude oil in rats

The embryotoxic potential of a Prudhoe Bay crude oil (PBCO) was investigated in rats. PBCO was administered orally to pregnant rats as (i) a single dose on various gestation days, (ii) a single variable dose on gestation day 6, or (iii) as daily doses from day 6 to day 17 of pregnancy. PBCO administered during the earlier stages of pregnancy (day 3, 6 or 11) but not during the later stages, affected the reproductive performance of pregnant rats by significantly increasing the number of resorptions including fetal death and by decreasing the fetal weight. A dose-dependent increase in fetomortality was also observed. Multiple exposure to low levels of crude oil also caused a significant reduction in maternal body weight besides other embryotoxic changes.     

Modulation of Th1/Th2 cytokine production by selective and nonselective phosphodiesterase inhibitors administered to mice

Phosphodiesterase (PDE) inhibitors can modulate the functions of immune cells, including T lymphocytes, due to increased intracellular levels of cyclic nucleotides. The drugs (aminophylline, milrinone and sildenafil) were administered once or five times at 24 h intervals at the following doses: 20 mg/kg, im, 1 mg/kg, im and 1 mg/kg, po, respectively.Th1 and Th2 cytokine levels (IL-2, IFN-γ, IL-4, IL-5, TNF) were determined 12, 24 or 72 h after the last administration of the drugs. A commercial BD^TM Cytometric Bead Array Mouse Th1/Th2 Cytokine Kit (CBA) was used to determine the levels of Th1/Th2 cytokines in the serum.Neither of the PDE inhibitors under investigation administered once changed IFN-γ, TNF and IL-4 production. A single dose of aminophyl-line decreased the production of IL-2 (after 12 h). A single dose of milrinone did not affect Th1/Th2 cytokine secretion. Sildenafil administered once decreased the production of IL-2 (after 72 h). A temporary enhancement in the level of IL-5 was observed 12 h after a single dose of sildenafil. No changes in Th1 and Th2 cytokine production were observed after five doses of PDE inhibitors under investigation. These results indicate that nonstimulated lymphocytes Th1 and Th2 exhibited a slight sensitivity to aminophylline and sildenafil. The drugs under investigation were ineffective inhibitors of Th1/Th2 cytokine production.     

Teratogenic potential of quercetin in the rat

A single oral dose of 2, 20, 200, or 2000 mg qucrcetin/kg was administered to pregnant rats on the morning of day 9 of gestation. Other groups of pregnant rats received similar oral doses of quercetin daily, on days 6–15 of gestation. Some quercetin-treated groups showed a significant decrease in the average weight of day-20 foetuses compared with the corresponding control weight. However, studies of the foetuses recovered on day 20 of gestation failed to reveal any reproducible dose-related syndrome of teratogenic effects attributable to quercetin treatment.     

The effects of perinatal/juvenile heptachlor exposure on adult immune and reproductive system function in rats.

This study was performed to determine if developmental exposure of rats to heptachlor (H) during the last half of gestation through puberty adversely affects adult functioning of the immune and reproductive systems. Time-bred pregnant female Sprague-Dawley rats were dosed by gavage with H (0, 30, 300, or 3000 microg/kg/day) from gestation day (GD) 12 to postnatal day (PND) 7, followed by direct dosing of the pups with H through PND 42. Separate groups of rats were evaluated with a battery of immune function tests, while other groups of rats were evaluated for reproductive development and function. Additional groups of rats were euthanized at the end of the dosing period for histological analyses of major organ systems. Some dams and PND 7 pups were euthanized; milk, plasma, fat and/or tissues were assayed for H and heptachlor epoxide B (HEB), a major metabolite of H. The amount of H and HEB found in milk, blood, fat, and tissues was proportional to the dose of H administered. There were no effects on the number or survival of pups born to H-exposed dams nor to pups exposed postnatally. There were no effects on the number of treated dams delivering litters or on litter size, nor were there any effects on any of the reproductive end points examined in the F(0) or F(1) rats. There were no effects of H exposure on lymphoid organ weights, splenic natural killer (NK) cell activity, and splenic lymphoproliferative (LP) responses to mitogens and allogeneic cells in a mixed lymphocyte response (MLR) assay at 8 weeks of age. H exposure did not alter delayed or contact hypersensitivity at 10 or 17 weeks of age, respectively. However, the primary IgM antibody response to sheep red blood cells (SRBCs) was suppressed in a dose-dependent manner in males, but not females, at 8 weeks of age. The percentage of B lymphocytes (OX12(+)OX19(-)) in spleen was also reduced in the high-dose males. The anti-SRBC IgM response was reduced only in males exposed to 30 microg H/kg/day in a separate group of rats 21 weeks of age. In these same rats, at 26 weeks of age, the secondary IgG antibody response to SRBCs was suppressed in all of the H-exposed males, but not females. These data indicate that perinatal exposure of male rats to H results in suppression of the primary IgM and secondary IgG anti-SRBC responses. Suppression of these antibody responses persisted for up to 20 weeks after the last exposure to H, at a total exposure of approximately 1500 microg H/kg/rat.     

Influence of various surface-active agents on the activity of several enzymes in the brush border of enterocytes.

Single doses of 25–70 mg/kg of lead nitrate were administered iv to pregnant rats on Days 8–17 of gestation (Day 1 = sperm found). Malformations were produced with lead when administered on Day 9 of gestation, producing a urorectocaudal syndrome of malformations. Lead nitrate was increasingly embryo- and fetotoxic when administered on later days of gestation (Days 10–15) but not teratogenic. Hydrocephalus and hemorrhage of the central nervous system were produced on Day 16 of gestation. On Day 16 and therafter, fetal toxicity (resorptions) sharply declined. Postnatal survival of newborn exposed in utero to lead on Day 9 of gestation was very poor. Significant quantities of lead were transferred into the fetus; however, the placenta appeared to greatly limit the passage of lead since large maternal-fetal concentration gradients existed.     

Potentiation of indomethacin-induced anti-inflammatory response by pioglitazone in carrageenan-induced acute inflammation in rats: Role of PPARγ receptors

This study aimed to assess the interaction between anti-inflammatory effects of pioglitazone (peroxysome proliferator activated receptor-gamma (PPARγ) agonist, PGL), and indomethacin (cyclooxygenase (COX) inhibitor, IND) and to evaluate the possible underlying mechanisms. Paw edema induced by carrageenan was used to induce inflammation. Different doses of IND (0.3–10 mg/kg) and PGL (1–20 mg/kg) alone or in combination were administered intraperitoneally to rats. Paw tissue levels of PPARγ, COX-2, and prostaglandin E2 and serum levels of TNF-α and IL-10 were also estimated.Doses of IND and PGL showed a statistically significant anti-inflammatory effect. Combination of a non-effective dose of IND (0.3 mg/kg) with increasing doses of PGL (1–10 mg/kg) resulted in potentiated anti-inflammation and vise versa. IND, PGL and the combination were able to reduce the COX-2, PGE2 contents and TNF-α level. Moreover, all these treatments caused elevation in PPARγ levels and IL-10 levels. However, when the rats were pre-treated with GW-9662 (a selective PPARγ antagonist), all the anti-inflammation and alterations in the biochemical factors were antagonized.These results showed that PGL markedly enhanced the anti-inflammatory activity of IND and this effect mediated partly at least, through PPARγ. Possible mechanisms of the interaction were that PGL stimulates the PPARγ and inhibits COX-2 by those cytokines that trigger the PPARγ and also inhibit COX-2. This study suggests that combination therapy with pioglitazone and indomethacin may provide an alternative for the clinical control of inflammation especially in patients with diabetes.     

Chromosomal aberrations induced by sodium nitrite in bone marrow of adult rats and liver cells of transplacentally exposed embryos

A commonly used food preservative, sodium nitrite, was administered in the drinking water to pregnant (d 5-18 of gestation) and nonpregnant albino rats. Sodium nitrite induced chromosomal aberrations in bone marrow of both pregnant and nonpregnant adults and liver of transplacently exposed embryos. The magnitude of the effect was greater in embryonic liver cells than in adult bone marrow cells.     

The New Salicylate Derivative UR-1505 Modulates the Th2/Humoral Response in a Dextran Sodium Sulphate Model of Colitis That Resembles Ulcerative Colitis

The aim of the present study was to evaluate the inmunomodulatory effects of UR-1505, a new salicylate derivative, on the T helper (Th)2 / humoral response produced during dextran sodium sulfate (DSS)-induced rat colitis. In the in vitro studies, UR-1505 (300 μM) inhibited both the production of interleukin (IL)-10 and IL-5 in concanavalin A (Con A)-activated splenocytes and the production of immunoglobulin (Ig) G and IgA by B-lymphocytes. However, in contrast to the in vitro results, the administration of UR-1505 (10 and 30 mg/kg per day) to rats with established DSS-colitis enhanced both IL-10 and IgA production, whereas it inhibited IgG production, thus ameliorating the intestinal inflammation.     

Prenatal treatment with clomipramine: effects on the behaviour of male and female adolescent rats

The tricyclic anti-depressant clomipramine (7.5 or 15 mg/kg/day) was administered to pregnant rats between days 8 and 21 of gestation. Between postnatal days 31 and 47, both male and female offspring received three behavioural tests. Prenatal clomipramine (15 mg/kg/day) increased baseline acoustic startle in females, but not in males; both sexes showed greater between-day response decrements if they had received clompramine. In the social interaction test of anxiety, males prenatally exposed to clomipramine (both dosages), and females prenatally exposed to 7.5 mg/kg/day, revealed a similar profile to that seen after chronic administration of benzodiazepines in the adult. The likelihood that differences in within-session habituation could underlie the changes in social interaction that have been found in this and other studies is assessed.     

商陆皂苷甲对大鼠Heymann肾炎的治疗作用及对细胞因子的影响

目的： 商陆皂苷甲(EsA)对大鼠Heymann肾炎(passive Heymann nephritis,PHN)的治疗作用及机理。方法： 大鼠sc Fx1A抗原的抗体制成自身免疫性肾小球肾炎,EsA 5,10和20 mg.kg^-1.d^-1 ip,连续14 d。结果： EsA可以显著减少肾炎大鼠尿蛋白的产生,电镜和免疫荧光检查发现EsA治疗后肾炎大鼠病理情况明显好转。EsA治疗对血清中炎性细胞因子肿瘤坏死因子（TNF）、白细胞介素1（IL-1）和白细胞介素6（IL-6）的产生具有显著的抑制作用。结论： EsA对大鼠Heymann肾炎具有显著治疗作用,抑制细胞因子产生可能参与EsA抗炎机制。     

敌枯双不同给药时间对妊娠Wistar大白鼠的致畸效应

本文主要讨论了化学药物不同给药时间的致畸效应。根据我们的实验,初步认为:对妊娠大鼠一次给药,以妊娠的第9、10或11天为宜,连续给药以9～12天为宜;如果必需进行整个妊娠期给药,可考虑在妊娠的6～15天连续给药。     

Genotoxic evaluation of pirfenidone using erythrocyte rodent micronucleus assay

Pirfenidone is a non-steroidal antifibrotic compound that has been proposed in clinical protocols and experimental studies as a pharmacological treatment for fibroproliferative diseases. The objective of this study was to determine the genotoxicity or cytotoxicity of three doses of pirfenidone using the micronuclei test in peripheral blood erythrocytes of rodent models. Pirfenidone was administered orally to Balb-C mice for 3 days, and also was administered topically to hairless Sprague Dawley rats during the final stage of gestation. Mice were sampled every 24 h over the course of 6 days; pregnant rats were sampled every 24 h during the last 6 days of gestation, and pups were sampled at birth. Blood smears were analyzed and the frequencies of micronucleated erythrocytes (MNEs), micronucleated polychromatic erythrocytes (MNPCEs), and the proportion of polychromatic erythrocytes (PCEs), were recorded in samples from mice, pregnant rats and rat neonates. Increases in MN frequencies (p < 0.03) were noted only in the positive control groups. No genotoxic effects or decreased PCE values were observed neither in newborn rats transplacentally exposed to pirfenidone, or in two adult rodent models when pirfenidone was administered orally or topically.     

Fetal development in alloxan-treated rats

The effect of alloxan on embryo and fetal development in rats was evaluated. Alloxan was injected intraperitoneally (ip) in pregnant rats at doses of 80 to 150 mg/kg at Day 0 (day of fertilization), and 110 mg/kg at Day 4 of pregnancy. Hyperglycemia was rarely produced at alloxan doses from 80 to 100 mg/kg, and the frequency of malformations observed was low. Higher doses (110 to 150 mg/kg) caused severe hyperglycemia, and maternal or embryonic death. When 110 mg/kg was administered on Day 4 of gestation (the day before embryo implantation), all rats had resorption nodules and litters with embryos with delayed growth. We recommend the induction of diabetes mellitus on Day 4 of pregnancy for studies of diabetes–gestation interaction.     

Ganoderma tsugae in vivo modulates Th1/Th2 and macrophage responses in an allergic murine model

We have reported that Ganoderma tsugae supplementation alleviates bronchoalveolar inflammation in an airway sensitization and challenge model with female BALB/c mice. However, the effects of G. tsugae supplementation in vivo on serum antibody levels, splenocyte and peritoneal microphage immune responses have not yet been determined. In this study, serum antibody levels, cytokines and splenocyte chemical mediators and peritoneal macrophage cultures from ovalbumin (OVA)-sensitized and -challenged mice were examined after continuously consuming G. tsugae supplementation diets for 5 weeks. The results showed that OVA sensitization and challenge significantly (P < 0.05) decreased the spontaneous production of IL-2 (Th1) cytokine, but significantly (P < 0.05) increased spontaneous and OVA-stimulated IL-4 (Th2) production in splenocyte cultures from experimental mice. OVA administration significantly decreased both spontaneous and LPS/IFN-γ-stimulated IL-1β and IL-6 levels in peritoneal macrophage cultures from experimental mice. However, dietary supplementation with G. tsugae significantly increased spontaneous IL-2 level, but slightly decreased spontaneous IL-4 level in cultured splenocyte supernatants in the experimental groups. G. tsugae supplementation enhanced pro-inflammatory cytokines IL-1β and IL-6 production in cultured peritoneal macrophages. However, the nitric oxide level from cultured peritoneal macrophages and serum OVA-specific IgE and IgG_2a antibody levels was not significantly affected. These results suggest that OVA sensitization and challenge induced a Th2-skewed splenocyte response and decreased peritoneal macrophage cytokine secretion. G. tsugae supplementation in vivo modulated the Th1/Th2 balance and enhanced macrophage immune responses. However, the supplementation diet could not fully reverse the Th2-skewed responses to level of Th1-skewed responses.