Histopathological Changes in Liver and Heart Tissue Associated with Experimental Ultraviolet Radiation A and B Exposure on Wistar Albino Rats.

This study aims to evaluate the influences of ultraviolet radiation A and B (UVA + B) exposure on the liver and heart organs of albino rats. Female Wistar Albino rats, whose hair of the dorsal skin was shaved, were exposed to a combined UVA + B radiation for 2 h/day, for 4 weeks in order to be compared with the control group. Histopathological findings in vital organs (liver and heart) were evaluated. Tissues were fixed in 10% buffered formalin (pH = 7.2) and embedded in paraffin. The histopathological findings were examined on the H&E stained sections with light microscopy. The results show that the liver and the heart were injured in the UVA + B group. Liver tissue in the UVA + B group showed minimal vacuolation, enlargement of hepatocytes and bile duct proliferation, and the heart tissue showed hibernomas; uniform large cells resembling brown fat with coarsely granular to multivacuolated cytoplasm that is eosinophilic or pale with a small central nucleus. The number of hibernoma cases was significantly higher in the UVA + B group compared with the control group (P = 0.021). The control group showed normal liver and heart histology with normal adipose tissue in the pericardium. As a result, UVA + B exposure has toxic effects, especially on the liver and the heart of Wistar albino rats. UV radiation may cause such adverse effects in humans. Therefore, protection against the harmful effects of UV radiation is of significant importance for skin and organs.

Combined treatment of sinapic acid and ellagic acid attenuates hyperglycemia in streptozotocin-induced diabetic rats.

In the present study, we aimed to investigate the effect of individual and combined treatment of sinapic acid (SA) and ellagic acid (EA) in streptozotocin (STZ)-induced diabetic rats. Rats were divided into eight groups (n = 7): Normal Control, Diabetic Control, Diabetic + Sinapic Acid, Diabetic + Ellagic Acid, Diabetic + Sinapic Acid + Ellagic Acid, Sinapic Acid, Ellagic Acid and Sinapic Acid + Ellagic Acid. Diabetic groups were injected with a single dose of 50 mg/kg STZ intraperitoneally. Rats received 20 mg/kg/day SA and 50 mg/kg/day EA intragastrically for 28 days. The numerical density of immunopositive ß-cells and volume density of pancreatic islets were calculated. Additionally, glucose and insulin levels in serum, MDA, GSH, and CAT levels of pancreatic tissue were measured. While serum glucose levels increased, serum insulin levels decreased in STZ-induced diabetic rats. But these changes in glucose and insulin were restored by individual and combined treatments of SA and EA. Also, individual and combined treatments of SA and EA increased insulin expression of ß-cells in STZ-induced diabetic rats. Moreover, these compounds improved deteriorating oxidative stress parameters in STZ-induced diabetic rats. Our study indicates that SA and EA, especially their combined treatments, can be used as an antihyperglycemic agent in diabetes.

The effect of maternal treatment with diclofenac sodium and thymoquinone on testicular parameters in rat offspring.

INTRODUCTION AND OBJECTIVE: Diclofenac sodium (DS) can have toxic effects on various tissues and organs, as well as causing foetal and new-born malformations. Thymoquinone (TQ), the basic bioactive compound of black seed oil, is an antioxidant and antineoplastic substance. The aim of our study was to explore the effects of DS and TQ exposure during gestation on offspring rat testicular histology. MATERIALS AND METHODS: Mother pregnant rats were divided into five groups: control, saline, DS, TQ and DS plus TQ (DS+TQ) four animals for each group. They were then treated as follows between day of 5 and 15 of gestation: the control group received no treatment. The saline group received physiological saline (1mg/kg/d) via the intraperitoneal (IP) route; the DS group received an intramuscular (IM) injection of DS (6.1mg/kg/d); the TQ group received TQ (5mg/kg/d) dissolved in drinking water; and the DS+TQ group received DS (6.1mg/kg/d) and TQ (5mg/kg/d) dissolved in water. After birth, the male rats were fed for four weeks, and at the end of this period offspring were sacrificed. Stereological methods, physical disector and Cavalieri principle were used for particle counting and volume estimation respectively. RESULTS: The results revealed a significant decrease in the total number of Sertoli and Leydig cells in 4-week-old rats in the DS group (p<0.05), and TQ not have provide protection against this adverse effect of DS. CONCLUSIONS: In this study, DS at a dose of 6.1mg/kg, equivalent to a dose of 1mg/kg in humans, decreased the number of Sertoli and Leydig cells, and TQ did not have a protective effect against the adverse effect of DS during the gestation period. These results show that new dose depend studies on TQ and DS interaction are requested to see protective effect of TQ.

A morphometric and stereological study on cervical spinal cord segment of goose.

In this study, volume density of white matter and grey matter areas of cervical segment of spinal cord in adult geese weighing 3-4 kg was examined using stereological methods. 10 geese were used as material without sex discrimination. All animals underwent perfusion with 10% buffered formaldehyde. Following the perfusion, animals were kept in 10% formaldehyde for 1 week. Geese were then dissected. Cervical area of spinal cord was revealed removing cervical spine. Tissue samples were obtained from each segment of cervical area. 5 µm thick cross-sections were taken from these tissue samples via microtome. Series of cross-sections were obtained by sampling in the ratio of 1/250 including 12 cross-sections from each cervical segment of every animal. Cross-sections were stained by haematoxylin eosin. They were photographed under microscope. Volume density (volume fractions) of both whole tissue and white matter and grey matter parts in each cervical segment of spinal cord were calculated using Cavalieri's Principle. In the study, total volume of cervical segment, volume of white matter and grey matter, and ratios of these volumes one another were assessed in goose.

Effects of spermine and the passive avoidance learning (PAL) following cerebral ischemia in chicks: Association with neuroprotection of pyramidal cells.

The aim of this study is to investigate the effects of spermine and the passive avoidance learning on hippocampus following transient cerebral ischemia in the chicks. The study is composed of the pure control (CG), sham (SG) and experimental groups (n=20). Experimental groups (ischemia group, IG and ischemia-spermine group, ISG) were exposed to ischemia for 20min whereas the SG was exposed to sham operation and CG group was not exposed to any operation. Passive avoidance learning (PAL) was applied to the half number of the subjects in each group. Both before and after 7days from the ischemia, operated animals were taken to PAL and then they were sacrificed. Total numbers of neurons in the hippocampus were stereologically estimated using Cresyl violet stained sections. We detected that number of neurons was increased following PAL and especially spermine treatment. According to our results, we suggested that spermine may reduce the deleterious effects of the ischemia by causing to increase in the neuronal number and so, it may be slightly supportive to the PAL.

Prenatal exposure to low-dose diclofenac sodium does not affect total neuron numbers in spinal segment T13 in rats.

The main purpose of the present study was to investigate the effects of diclofenac sodium (DS) on the total number of neurons in segment T13 of the spinal cord of offspring of pregnant rats using stereological methods. Eighteen adult female Wistar albino rats weighing 150-200g were used. Pregnant female rats were divided into three groups; a control group, a sham group and a DS (1mg/kg, intramuscular) exposed group. The DS and sham groups received injection from the 5th day of gestation to the 19th. Twenty eight days after birth, the offspring rats were perfused with 4% buffered formalin. T13, which is one of transverse spinal cord segments, were isolated and processed for routine paraffin histology. 5µm sections were obtained using a rotary microtome according to systematic random sampling strategies. Every 40th section was taken and sections were stained with modified Giemsa. All types of motor neuron cell were identified according to their morphology. In this study, the "disector-Cavalieri combination" method was used in the stereological examination of neurons. The motor neurons were counted in the right gray matter of the ventral horn in the spinal cord segment. The Kruskal-Wallis test was used for comparison the groups. In terms of motoneuron number, no significant difference among the groups was found (p>0.05). In conclusion, our results indicated that prenatal exposure to DS has no effect on the total number of motor neuron of the offspring rats.

Investigation of role of vascular endothelial growth factor, Annexin A5 and Apelin by immunohistochemistry method in placenta of preeclampsia patients.

Preeclampsia is a disease characterized by hypertension and proteinuria occurred after 20 weeks of gestation. Preeclampsia is a major cause of maternal and fetal morbidity and mortality.  The pathophysiological mechanism of preeclampsia is not known exactly yet. Preeclampsia endothelial cell dysfunction, associated with inadequate trophoblastic invasion is characterized by abnormal placentation. Vascular endothelial growth factor (VEGF) according to is an angiogenic cytokine, Annexin A5 is among endogenous peptides are both expressed from placental trophoblasts and Apelin is a multifunctional peptide and expressed by placental trophoblasts and endothelial cells. It was aimed to investigate roles of these parameters occurring in preeclampsia and to compare immunoreactivity of them in normal and preeclamptic placenta. In this study, placentas were collected from 20 normotensive pregnant women as controls, 16 mild-preeclamptic pregnant women, and 16 severe preeclamptic women. VEGF, Annexin A5 and Apelin were examined in samples of placenta tissues by streptavidin-biotin-peroxidase complex immunohistochemical methods. Immunoreactivity scores (IRS) were obtained for each parameter.  VEGF and Apelin IRS were increased significantly in preeclamptic groups compared with control group (p <0.026, p<0.002 respectively). But Annexin A5 IRS was decreased significantly in preeclamptic groups compared with control group (p<0.04). In correlation with the intensity of disease, increase in VEGF and Apelin, and decrease in Annexin A5 supports roles of hemo-dynamic alterations in fetoplacental circulation and structural alterations in uteroplacental bed occurring in preeclampsia.

Resveratrol Treatment Prevents Hippocampal Neurodegeneration in a Rodent Model of Traumatic Brain Injury.

AIM: Traumatic brain injury (TBI) is a complex process. Increasing evidence has demonstrated that reactive oxygen species contribute to brain injury. Resveratrol (RVT) which exhibits significant antioxidant properties, is neuroprotective against excitotoxicity, ischemia, and hypoxia. The aim of this study was to evaluate the neuroprotective effects of RVT on the hippocampus of a rat model of TBI. MATERIAL AND METHODS: Twenty eight rats were divided into four groups. A moderate degree of head trauma was induced using Feeney"s falling weight technique. Group 1 (control) underwent no intervention or treatment. Head trauma was induced in Group 2 (trauma) and no drug was administered. Head trauma was induced in Group 3 and low-dose RVT (50 mg/kg per day) was injected. In Group 4, high-dose RVT (100 mg/kg per day) was used after head trauma. Brain tissues were extracted immediately after perfusion without damaging the tissues. Histopathological and biochemistry parameters were studied. RESULTS: Brain tissue malondialdehyde (MDA) levels in the trauma group were significantly higher than those in the control, lowdose RVT-treated, and high-dose-RVT-treated groups. The superoxide dismutase (SOD) levels in the control group were significantly higher than those in the trauma, low-dose RVT-treated, and high-dose RVT-treated groups. Glutathione peroxidase (GSH-Px) levels in the control group were significantly higher than those in the trauma and low-dose RVT-treated groups. The level of oxidative deoxyribonucleic acid (DNA) damage (8-OHdG/106 dG) in the trauma group was higher than that in the control group, low-dose RVT-treated, and high-dose RVT-treated groups. CONCLUSION: Resveratrol has a healing effect on neurons after TBI.

The Effect of the Active Ingredient Thymoquinone on Flap Viability in Random Pattern Flaps in Rats.

Thymoquinone (TQ) is a plant extract that has been shown to have antioxidant, anti-inflammatory, angiogenic, antimicrobial, and anticarcinogenic effects. The aim of this study is to research how the use of TQ affects flap viability. 42 rats were placed into 6 groups, with 7 rats in each. A 3 × 10 cm McFarlane flap model was used on the test animals. The sham group had used neither surgical nor TQ treatment. The control group had surgery but no treatment afterwards. The preoperative TQ group was given oral doses of 2 mg/kg. TQ for 10 days preoperatively with no treatment after the surgical procedure. The postoperative TQ group received oral doses of 2 mg/kg TQ for 10 days after the surgical process. The preoperative + postoperative (pre + postoperative) TQ group was given oral doses of 2 mg/kg TQ for 10 days both preoperatively and postoperatively. Finally, the dimethylsulfoxide group received 10 mg/kg dimethylsulfoxide (DMSO) for 10 days both preoperatively and postoperatively. Ten days after surgery the findings were evaluated. The average rates of necrosis were found to be 29.7 % in the control group, 19.18 % in the preoperative TQ group, 13.05 % in the postoperative TQ group, 8.42 % in the pre + postoperative TQ group, and 29.03 % in the DMSO group. The experimental groups had better area measurement, histopathological, and electron microscopic results than the control group (All; p < 0.05). We believe that, because of its antioxidant, anti-inflammatory, and angiogenic properties, thymoquinone is an agent that can prevent ischemia-reperfusion damage and, therefore, prevent necrosis.

The acute effects of thymoquinone on acute peripheral nerve injury: an experimental study.

BACKGROUND: The purpose of this study was to evaluate the acute effects of thymoquinone (TQ) on acute nerve injury. METHODS: A rat model of crush injury of the sciatic nerve was used. Animals were divided into 3 groups: control, trauma, and TQ treatment groups (n=6 per group). Seven days after injury, sciatic nerve specimens were obtained from the site of the injury and analyzed histologically and stereologically. Axon diameter, myelin thickness, and axon density were measured. RESULTS: There were no significant differences in axon diameter, myelin thickness, or axon density among groups. CONCLUSION: TQ has no acute therapeutic effect on acute nerve injury.

Effects of ozone pretreatment on viability of random pattern skin flaps in rats.

BACKGROUND: Medical ozone is a chemical agent that consists of three oxygen atoms and has antioxidant, angiogenic and vasodilator effects. This study evaluated the effects of medical ozone pre-treatment on flap survival. MATERIALS AND METHODS: Rats were divided into four groups of 10 rats each and a 9 × 3 cm McFarlane flap was used. Sham group: Neither surgical nor ozone pretreatment was used. CONTROL GROUP: No pretreatment was used after surgery. Preoperative ozone group: Preoperative 1 mg/kg ozone was given intraperitoneally for 7 days. No pretreatment was used after surgery. Postoperative ozone Group: Postoperative 1 mg/kg ozone was given intraperitoneally for 7 days. After postoperative 1 week, all groups were evaluated by surface area measurement, histopathology and electron microscopy. RESULTS: With the experimental McFarlane flap model, the experimental groups had better surface area measurements, along with histopathological and electron microscopic results when compared with the control group. CONCLUSION: Medical ozone had positive effects on flap survival due to its antioxidant, angiogenic and vasodilator qualities.

Determination of micronutrients and oxidative stress status in the blood of STZ-induced experimental diabetic rat models.

This study aims to research the effect of streptozotocin (STZ) at different doses on the serum micronutrients and oxidative stress status in diabetic rat models. Twenty male rats averaged 250 g and 3-4 months old were used as experimental models. They were put in four groups composed of five rats each. Diabetic was induced by administering STZ 55 and 65 mg/kg intraperitonally. The serum micronutrients including minerals and vitamins (Cu, Zn, Mg, Fe, vitamins D, E, and C) and oxidative stress (malondialdehyde, MDA) were determined. Cu, Zn, and Vitamin D3 levels were found to increase significantly in STZ groups (p < 0.005). Retinol levels decreased significantly in STZ groups (p < 0.005). In the groups administered 55 mg/kg STZ ferrum and vitamin C levels were found significantly lower than the other groups (p < 0.005). In the group given 65 mg/kg STZ α-tocopherol levels were highest (p < 0.005) among other groups. There was not any difference between the groups for MDA, Cu/Zn, and Mg. For both doses, oxidative stress status was not significantly affected within 48 h of the application, however, some micronutrients were affected significantly.

The effect of prenatal exposure of a non-steroidal anti-inflammatory drug on the optic nerve of female rats: a stereological, histological, and electron microscopic study.

OBJECTIVE: Non-steroidal anti-inflammatory drugs (NSAIDs) can have adverse effects for in both mother and fetus following administration during the prenatal period. If given during pregnancy, diclofenac sodium (DS), an NSAID, is given during the pregnancy, may also affect the development of the central nervous system (CNS) or related structures. METHODS: Pregnant rats were separated into pure control (PG), saline (SG) and diclofenac groups (DG). A daily dose of 1 mg/kg of DS and 1 mL/kg saline was injected intraperitoneally to the DG and SG groups, respectively, from the 5th gestation day for a 15 day of period; the PG group received no treatment. After spontaneous delivery, female offspring were obtained from all groups. After the 20th week of postnatal life, the animals (n = 6 for each group) were perfused and the right optic nerves were resected. Sections were subjected to stereological and histological analysis. RESULTS: There were no significant differences (p > 0.05) between PG, SG and DG groups with respect to myelin thickness, axonal cross-sectional area, axon numerical density, total section area of optic nerve and axon number. CONCLUSIONS: Histological and stereological results indicated that treatment with DS or saline produced undesirable effects on female rat optic nerve development and myelinization with respect to morphology.

Effect of prenatal exposure to diclofenac sodium on the male rat arteries: a stereological and histopathological study.

In this study, we investigated the morphometric and histological alterations of the aorta, brachial, and femoral arteries in 4- and 20-week-old rats that were prenatally exposed to diclofenac sodium (DS). For this purpose, pregnant rats were divided into three groups: control, saline injected, and drug treated. Beginning from day 5 after mating through day 15 of pregnancy, saline or DS (1 mg/kg daily) was intraperitoneally injected into groups 2 and 3. No injection was given to the rats in the control group. After spontaneous delivery, male offspring were obtained. At the end of weeks 4 and 20, vessel samples were removed. After dissection and routine histological preparation, histopathological and stereological investigations were made. Our results indicate that both saline and DS application lead to a decrease in the mean volume fraction of tunica media in all vessel walls, but result in an increase of the same fraction of lumen to the whole vessel wall, especially in 4-week-old rats. Elastic fibers of the vessel wall were affected by DS treatment, because a decrease of the elastic fiber was observed in this group. Finally, in light of our findings, we suggest that DS or saline may lead to vascular changes (i.e., vasodilatation or vasoconstriction) in rats that are prenatally subjected to increased volume of maternal blood resulting from injection.

Total neuron numbers in CA1-4 sectors of the dog hippocampus.

BACKGROUND & OBJECTIVES: Early reports addressed morphological asymmetry in the cross-sectional width of the rat hippocampus. The present study was aimed at counting total number of neurons in CA1-4 sectors and the subiculum of the dog hippocampus as well as investigating possible left /right and male/female asymmetry. METHODS: Adult mongrel dogs (8 female and 5 male) were assessed by the right and left pawedness and sacrificed by exsanguinations. In each hippocampus dissected, the total neuron numbers of CAs and subiculum were estimated by the physical fractioning method. RESULTS: Significant hemispheric asymmetries were found in the number of pyramidal cells of CA1, CA3/2, CA4 and the subiculum. Sex difference was also found in the subiculum, in favour of the males. INTERPRETATION & CONCLUSION: Our study indicated a left dominant asymmetry in males and right dominancy in females as well as no functional asymmetry in specific regions of the dog hippocampus. Further investigations are necessary to verify the hypothesis that hippocampal morphological asymmetries in normal subjects are functionally related in memory or in cognitive skills.

Prenatal diclofenac sodium administration increases the number of Purkinje cells in female rats: a stereological study.

Diclofenac sodium (DS) may affect the number of Purkinje cells in the developing cerebellum since DS can easily be transported from the maternal to the fetal physiological system during the pregnancy. In the present study, the effects of prenatal exposure to DS on the number of Purkinje cells in the cerebellum of 4-week-old (4W-old) and 20-week-old (20W-old) female rats were investigated. There were two main groups: the drug-treated group (DTG) and the control group (CG). Beginning from the 5th day after mating for a period of 15 days, a daily dose of 1 mg/kg of DS (Voltaren, 75 mg/3 ml ampul, Novartis, Mefar Ilaç Sanayi A.S., Kartal, Istanbul, Turkey) was intraperitoneally injected in the DTG of pregnant rats. In contrast, a daily dose of 1 ml/kg of isotonic saline was intraperitoneally administered to the CG of pregnant rats during the same period. After spontaneous delivery, female offspring were obtained, and the main groups' offspring were divided into two subgroups as a 4W-old group and a 20W-old group. Therefore, there were four groups at the end of the experiment: the 4W-old DTG and the CG, and the 20W-old DTG and the CG. At the end of 4W and 20W, offspring were perfused, their brains were dissected, and the number of cells estimated via the optical fractionator technique. Our results showed that while the total number of Purkinje cells in the cerebellum of offspring of DT 20W-old female rats was significantly higher than that of the CG, there was no significant difference between the 4W-old DTG and the control groups. Therefore, it could be suggested that DS administration during the prenatal period increases the number of Purkinje cells in the cerebellum of a developing female rat throughout postnatal 20W.

Effect of prenatal exposure to mobile phone on pyramidal cell numbers in the mouse hippocampus: a stereological study.

Because of the possible risk factor for the health, World Health Organization (WHO) recommended the study with animals on the developing nervous system concerning the exposure to radiofrequency (RF) field. A few studies related to hippocampal exposure are available, which indicate the impact of RF field in some parameters. The present study investigated the effect of exposure to mobile phone on developing hippocampus. Male and female Swiss albino mice were housed as control and mobile phone exposed groups. The pregnant animals in tested group were exposed to the effects of mobile phone in a room possessing the exposure system. The left hemispheres of the brains were processed by frozen microtome. The sections obtained were stained with Hematoxylin & Eosin. For cell counting by the optical fractionator method, a pilot study was first performed. Hippocampal areas were analyzed using Axiovision software running on a personal computer. The optical dissector, systematically and randomly spaced, was focused to the widest profile of the pyramidal cell nucleus. No significant difference in pyramidal cell number of total Cornu Ammonis (CA) sectors of hippocampus was found between the control and the mobile phone exposed groups (p > .05). It was concluded that further study is needed in this field due to popular use of mobile telephones and relatively high exposure to the developing brain.

Prenatal exposure to a non-steroidal anti-inflammatory drug or saline solution impairs sciatic nerve morphology: a stereological and histological study.

The toxic effect of non-steroidal anti-inflammatory drugs (NSAIDs) during development has been widely investigated. While it has been shown that these drugs impair central nervous development and compromise the neural activity, the effects of these substances on the development of peripheral nerves are still not clarified. In the present study, sciatic nerves withdrawn from three experimental groups of 4-week-old rats, prenatally exposed to either saline solution, or diclofenac sodium, and controls not exposed to any substance, were evaluated in terms of axon number, cross-sectional area of axon and myelin sheet thickness as well as of the ultrastructure of nerve fibers. Comparisons of stereological estimations among these three groups showed that axon number and mean axon cross-sectional area, but not average myelin sheet thickness, were significantly decreased in rats that were exposed to both diclofenac sodium and also to the saline solution, in comparison of the control group. Electron microscope analysis revealed, in both treated groups, deterioration of myelin sheaths that was more pronounced in rats that were exposed to diclofenac sodium. Altogether, these findings show that the prenatal administration of both diclofenac sodium and saline solution impairs peripheral nervous system development, thus suggesting that this potential teratogenic effect should be also taken into consideration in the clinical use of these substances in pregnant patients.

Effect of prenatal exposure to diclofenac sodium on Purkinje cell numbers in rat cerebellum: a stereological study.

Diclofenac sodium (DS) is commonly used as a non-steroidal anti-inflammatory drug. Although several adverse effects are clearly established, it is still unknown whether prenatal exposure to DS has an effect on the development of the cerebellum. In this study, we investigated the total number of Purkinje cells of the cerebellum in a control group and in a DS-treated group of male rats using a stereological method. The DS in a dose of 1 mg/kg daily was intraperitoneally injected to the drug-treated group of pregnant rats beginning from the 5th day after mating for a period of 15 days during pregnancy. Physiological serum at 1 ml dose was intraperitoneally injected to the control group of pregnant rats at the same period. After delivery, male offspring were obtained and each main group was divided into two subgroups that were 4-week-old (4W-old) and 20-week-old (20W-old). Our results showed that the total number of Purkinje cells in offspring of drug-treated rats was significantly lower than in the offspring of control animals. These results suggest that the Purkinje cells of a developing cerebellum may be affected by administration of DS during the prenatal period.

Effect of prenatal exposure to an anti-inflammatory drug on neuron number in cornu ammonis and dentate gyrus of the rat hippocampus: a stereological study.

Prenatal exposed to an anti-inflammatory drug is a major problem for the developing central nervous system. It is not well known the effect of prenatal exposed to a non-steroidal anti-inflammatory drug on the hippocampus. Total neuron number in one side of the cornu ammonis (CA) and gyrus dentatus (GD) of the hippocampal formation in control and drug-treated (diclofenac sodium, DS) groups of male rats was estimated using the optical fractionator technique. Each main group has also two subgroups that are 4 weeks old (4W-old) and 20 weeks old (20W-old). In CA, no significant difference between 4W-old DS-treated and their control was found, but a significant difference was observed between 20W-old DS-treated and their controls. A decreasing of neuron number was 12% for 20W-old DS-treated group. In GD, a decreasing of the granule cell number in 4W-old of DS-treated group was seen but an increasing of granule cell number was found in the 20W-old drug-treated rats in comparison to its control group, 7% and 9%, respectively. Although an increasing of neuron number in CA at the control group was seen with age, from 4th week to 20th week (10%), age-dependent substantial granule cell decline (17%) was observed in GD. No age effect on the total cell numbers of CA and GD of the drug-treated groups was seen in comparison to 4W-old week and 20W-old. A pronounced neuron loss observed in the drug-treated group may be attributed to the neurotoxicity of diclofenac sodium (DS) on the developing hippocampal formation. Age-dependent neuron increase in the CA of 20W-old and neuron decline in GD of 20W-old control groups may be a result of a dual effect of saline injection during the fetal life, since these animals were exposed to a stress of 15-day-period of saline injection, prenatal stress. The reason of no age effect on CA and GD cell number in the drug-treated groups may be attributed to the depletion of the progenitor cells due to neurotoxicity of DS in the fetal life of these animals.