Modulatory effects of quercetin on histological changes, biochemical and oxidative stress of rat placenta induced by inhalation exposure to crude oil vapor.

The inhalation exposure to crude oil vapor (COV) has been shown to have adverse effects on the placenta and fetal development. The modulatory effects of quercetin (QUE) as a natural phenolic compound with antioxidant properties are promising for the protection of placental structure. This study aimed to investigate the modulatory role of QUE in mitigating histopathological damage, oxidative stress, and biochemical alteration in the placenta of COV-exposed pregnant rats. Forty-eight pregnant rats were divided into eight groups (days 15 and 20) as follows: 1-2) Control groups, 3-4) COV groups, 5-6) COV+QUE groups, and 7-8) QUE-treated groups (50 mg/kg). The inhalation method was used to expose pregnant rats to COV, and QUE was administered orally. On the 15th and 20th days of gestation, placental tissue was analyzed using PAS and H&E staining and immunohistochemistry. The expression of the caspase-3 gene and oxidative stress biomarkers including TAC, CAT, MDA, GPx, and SOD were investigated in the placental tissue. The COV significantly decreased the weight, diameter, and thickness of the placenta as well as the thickness of the junctional zone and labyrinth and the number of trophoblast giant cells in 15- and 20-day-old placentas (P<0.05). Also, COV significantly increased placental expression of caspase-3 and the oxidative stress biomarkers (P<0.05). The administration of QUE along with exposure to COV reduced morphometric and histological alteration, oxidative stress, and caspase-3 expression (P<0.05). Our findings indicated that QUE in COV-exposed pregnant rats can prevent placental histopathological alternations by increasing the activity of the antioxidant system.

Protective effect of quercetin on fetal development and congenital skeletal anomalies against exposure of pregnant Wistar rats to crude oil vapor.

BACKGROUND: Epidemiological evidence indicates a relationship between maternal exposure to crude oil vapors (COV) during pregnancy and adverse pregnancy outcomes. Quercetin (QUE) is a plant flavonoid with purported antioxidant and anti-inflammatory effects, which has been shown to prevent birth defects. This study was aimed to investigate the protective role of QUE on fetal development and congenital skeletal anomalies caused by exposure of pregnant rats to COV. METHODS: Twenty-four pregnant Wistar rats were randomly categorized into four groups of control, COV, COV + QUE, and QUE (50 mg/kg). The inhalation method was used to expose pregnant rats to COV from day 0 to 20 of pregnancy, and QUE was administered orally during this period. On day 20 of gestation, the animals were anesthetized and a laparotomy was performed, and then the weight and crown rump length (CRL) of the fetuses were determined. Skeletal stereomicroscopic evaluations of fetuses were performed using Alcian blue/Alizarin red staining method, and the expression of osteogenesis-related genes (Runx2 and BMP-4) was evaluated using qPCR. RESULTS: This study showed that prenatal exposure to COV significantly reduced fetal weight and CRL, and expression of Runx2 and BMP-4 genes. Moreover, COV significantly increased the incidence of congenital skeletal anomalies such as cleft palate, spina bifida and non-ossification of the fetal bones. However, administration of QUE with exposure to COV improved fetal bone development and reduced congenital skeletal anomalies. CONCLUSION: QUE can ameliorate the teratogenic effects of prenatal exposure to COV by increasing the expression of osteogenesis-related genes.

Prenatal exposure to crude oil vapor reduces differentiation potential of rat fetal mesenchymal stem cells by regulating ERK1/2 and PI3K signaling pathways: Protective effect of quercetin.

It has been indicated that crude oil vapor (COV) induces tissue damage by several molecular mechanisms. Quercetin (QT) as an important component of food with antioxidant properties has a protective role against cell toxicity caused by many pollutants. However, data related to the adverse effects of crude oil vapor (COV) on stem cell fate and differentiation and the role of quercetin (QT) in protecting stem cells against the toxicity caused by these pollutants is very limited. This study aimed to explore the protective effect of QT against the adverse effects of COV on fetal mesenchymal stem cells (fMSCs) differentiation. Twenty-four pregnant Wistar rats were categorized into 4 groups including the control, COV, COV+QT, and QT. Rats were exposed to COV from gestational day (GD) 0-15 and received QT by gavage. The fMSCs were isolated from fetuses, and cell proliferation, differentiation potential, expression of osteogenesis and adipogenesis-related genes, and phosphorylation of PI3K and ERK1/2 signaling proteins were evaluated. The results showed that COV reduced the proliferation and differentiation of fMSCs through the activation of PI3K and ERK1/2 signaling pathways. Also, COV significantly decreased the expression of osteonectin, ALP, BMP-6, Runx-2, PPARγ, and CREBBP genes in differentiated cells. QT treatment increased the proliferation and differentiation of fMSCs in COV-exposed rats. In conclusion, our findings suggest that prenatal exposure to COV impaired fMSCs differentiation and QT reduced the adverse effects of COV by regulating ERK1/2 and PI3K signaling pathways.

Inhalation exposure to crude oil vapor induces behavioural deficits by increasing oxidative stress and histopathological changes in rat hippocampus: Quercetin therapeutic approach.

OBJECTIVE: Inhalation exposure to crude oil vapor (COV) and petroleum products is considered responsible for neurobehavioral toxicity in human and animal models. The antioxidant activity of quercetin (Que) and its derivatives are promising for protecting the hippocampus. This study aimed to evaluate the neuroprotective role of Que against COV-induced behavioral alterations and hippocampus damage. METHODS: Eighteen adult male Wistar rats were randomly divided into the following three groups (n = 6): the control, the COV, and the COV + Que group. The inhalation method was used to expose the rats to crude oil vapors for 5 h daily, and Que (50 mg/kg) was administered orally. After 30 days of treatment, the spatial working memory and anxiety levels were evaluated using the cross-arm maze and elevated plus maze (EPM), respectively. TUNEL assay and hematoxylin-eosin (H&E) staining were used to identify the necrosis, normal and apoptotic cells in the hippocampus. Moreover, the levels of oxidative stress biomarkers including malondialdehyde (MDA), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), and total antioxidant capacity (TAC) were investigated in the hippocampus tissue. RESULTS: The results indicated that exposure to COV was associated with a significant decrease in spatial working memory and activity of CAT, TAC, SOD, and GPx enzymes compared to the control (P < 0.05). Moreover, COV significantly increased the level of anxiety, MDA, and hippocampal apoptosis (P < 0.05). The simultaneous administration of quercetin along with exposure to COV improved the behavioral alterations, activity of antioxidant enzymes, and hippocampal apoptosis. CONCLUSIONS: These findings suggest that quercetin prevents COV-induced hippocampal damage by enhancing the antioxidant system and preventing cell apoptosis.

Paravertebral brachial plexus block in sheep: a cadaveric and in vivo study.

The Objective of the present study was to evaluate paravertebral brachial plexus block in sheep. A group of 13 healthy sheep with 20.10 ± 2.20 kg weight and five months of age were used. In phase I, in five sheep, an insulated needle attached to a nerve stimulator was directed to the location of C6, C7, C8 and T1 nerves and a 1.50 mL of a solution containing 1:1 methylene blue 1.00% and lidocaine 1.00% was injected at each site. Then, the cervical and thoracic areas were dissected and assessed in the cadavers. In phase II, cervical paravertebral block with 2.00% lidocaine and subsequent assessments were done in eight live sheep. Cadaveric evaluations revealed dye spread in C6 to T1 nerves: 61.75 ± 5.50, 72.75 ± 9.18, 40.75 ± 2.99 and 18.75 ± 3.30 mm, respectively. In three sheep, dye distribution in the anterior mediastinum was observed. In phase II, the onsets of anesthesia were determined within 10 and 15 min for sensory and motor blocks, respectively. Anesthesia at axillary, musculocutaneous, radial and ulnar skin sites and motor block lasted for 67.50 ± 15.80, 63.70 ± 16.00, 55.00 ± 21.70, 56.70 ± 19.70 and 76.40 ± 24.30 min, respectively. In three sheep, no anesthesia was observed for radial and ulnar skin sites. In conclusion, paravertebral brachial plexus block in sheep provided an acceptable block for the upper parts of the elbow joint, however, it was not effective and reliable for more distal structures.

Effect of bromelain on sperm quality, testicular oxidative stress and expression of oestrogen receptors in bisphenol-A treated male mice.

Bisphenol A (BPA) as an endocrine-disrupting chemical (EDC) with low estrogenic activity increases oxidative stress and testicular damage. Bromelain is a mixture of different thiol endopeptidases and other components with many uses as a natural anti-inflammatory enzyme. The present study aimed to evaluate the effect of bromelain on male reproductive failure induced by BPA. A total of 60 healthy adult male mice were randomly divided into six groups (n = 6), including control, bromelain (70 mg/kg), BPA (5 and 600 mg/kg), and BPA (5 and 600 mg/kg) + bromelain. BPA and bromelain were administrated orally for 35 days. Then, the epididymis and testes were removed to evaluate sperm parameters, oxidative stress markers, serum levels of testosterone concentrations, and oestrogen receptors expression. The BPA significantly (P < 0.05) decreased sperm count, motility, viability, and normal sperm morphology, as well as testosterone levels, oestrogen receptors alpha (ERα) and beta (ERß), GPx, CAT, and SOD activity than control. Also, BPA significantly (P < 0.05) increased the sperm anomalies, and MDA concentration. Co-administration of bromelain + BPA caused a significantly (P < 0.05) increase sperm count, normal sperm morphology, testosterone levels, expression of ERα and ERß, and GPx, CAT, and SOD activity than the BPA group (P < 0.05). Also, Bromelain significantly (P < 0.05) decreased sperm anomalies and MDA concentration than control. Based on the results of this study, it appears that BPA causes side effects on male reproduction. While, bromelain has the potential to reduce the side effects of BPA on the male reproductive system.

Potential protective effect of quercetin on the male reproductive system against exposure of Wistar rats to crude oil vapor: Genetic, biochemical, and histopathological evidence.

Toxic compounds in crude oil vapor (COV), including polycyclic aromatic hydrocarbons (PAHs), are associated with adverse effects on reproduction in living organisms. Quercetin (QT) is the most plentiful flavonoid in vegetables and fruits, with antioxidant activities. This study aimed to evaluate the protective role of QT on testicular toxicity induced by COV. Twenty-four adult male Wistar rats were randomly divided into four groups (n = 6) including control, quercetin (QT) (50 mg/kg), crude oil vapors (COV), and COV + QT. The inhalation method was used to expose the rats to crude oil vapors for 5 h daily, and QT was administered orally. After 30 days, the rats were euthanized, then, the testes were removed for gonadosomatic index (GSI), sperm parameters, H&E staining, the activity of the antioxidant enzymes, and apoptotic gene expression assessments. The COV statistically significantly (P < 0.05) reduced GSI, sperm count, motility, viability, and sperm normal morphology, histological indexes, and antioxidant enzyme activities than control. Also, COV statistically significantly (P < 0.05) increased the expression of caspase-3, p-53, and Bax genes and decreased Bcl-2 gene expression. Co-administration of QT + COV caused a statistically significant (P < 0.05) decrease in Bax gene expression and increased antioxidant enzyme activities, Bcl-2 gene expression, and reproductive parameters than the COV group. Based on the results of this study, it appears that crude oil vapor causes side effects on male reproduction. Yet, quercetin has the potential to reduce the side effects of crude oil vapor on the male reproductive system.

Dimorphic evaluation of hippocampal changes in rat model of demyelination: A comparative functional, morphometric, and histological study.

BACKGROUND: Multiple sclerosis (MS) is the most common autoimmune disease. Progressive depletion of the brain and spinal cord tissue appears at the onset of the disease. Several studies have shown the increased size of the ventricles of the brain and decreases in the area of the corpus callosum and the width of the brain. Other important symptoms of this disease are cognitive, learning, and memory disorders. AIM: The aim of this study was to compare morphometric, histological, and functional changes in the demyelination model in both sexes. MATERIALS AND METHODS: In this experimental study, male and female Wistar rats were studied in four experimental groups. Demyelination was induced by the injection of ethidium bromide in the ventricular region. The chronic effect of demyelination on spatial memory, movement, and coordination was investigated using the Morris Water Maze (MWM), and clinical and balance beam tests, respectively. Myelin degradation, cell death and neurogenesis were estimated using Luxol Fast Blue staining and immunohistochemistry (Caspase-3 and Nestin markers). In addition, morphometric findings were recorded for the brain and hippocampus (weight, volume, length, width). RESULT: Demyelination increased the time and distance index and decreased the residence time in the target quarter in the water maze test (p < .001). It also increases the neuromuscular and modified neurological severity score (p < .01). Demyelination increases caspase-3 (p < .05) expression and decreases Nestin expression (p < .001), which are directly related to the extent of damage. CONCLUSION: This study showed an interaction between hippocampal structural and functional networks in explaining spatial learning and memory in the early stages of MS.

Effects of omega-3 fatty acid supplementation during chronic maternal hypoxia on behavioral disorders in male rat offspring: the role of Trk family and oxidative stress.

Maternal hypoxia due to a lack of blood flow and insufficient oxygen supply in the brain leads to behavioral disorders in adult offspring. Fish oil includes docosahexaenoic acid (DHA), a significant component of membrane phospholipids of nerve cells, which improved cognition, and memory. Trk family receptors are activated by hypoxic induction factor (HIF), and are involved in the neurotrophin's protective effects at the cellular level. Here we studied the biochemical, and molecular mechanisms of the protective effect of fish oil during the chronic maternal hypoxia model on behavioral responses in male rat offspring. Pregnant female rats were randomly divided into 4 experimental groups: 1) ctr; Control rats were pregnant 2) Hyp; Pregnant female rats received hypoxia from 6 to 15th day of pregnancy, with 10% oxygen intensity, and 90% nitrogen; 3) FO; Pregnant female rats received fish oil (F8020 1 ml / day, for ten consecutive days Orally), and 4) FO / Hyp; Pregnant female rats received hypoxia plus fish oil in the same manner. Behavioral parameters were evaluated in 28-day-old male offspring. HIF-1α, TrkB, and P75 gene expression were measured in the offspring's brain. Maternal hypoxia impaired memory performance, and locomotor activity in offspring. Besides, Trk family gene expression, and oxidative stress indicators showed a significant increase in the offspring's brain exposed to maternal hypoxia compared to the control group. Overall, fish oil improved behavioral parameters by inhibiting oxidative stress, and the expression of Trk family receptors.

Age estimation of Arabian mares by incisors morphometry and dentition changes.

Accurate estimation of a horse's age based on the condition of the tooth status is necessary as a scientific and artistic technique, which has not been performed so far in pure Arabian horses of Khuzestan (southern west of Iran). This study aimed to investigate the age-dependent changes in the morphology and morphometry of the incisors in Arabian mares of Khuzestan to estimate age and to compare the estimated dental age and actual age. In this study, 78 Arabian mares of Khuzestan were examined in several equestrian clubs. Then, images were taken with a digital camera from the vestibular and occlusal surfaces of the lower incisors. Parameters of deciduous and permanent teeth eruption and their number, occlusal surface changes in the lower incisors including appearance and disappearance of the cup, enamel spot, dental star and appearance, and changes of Galvayne's groove in the upper corner incisor were investigated. Comparison of the clinical crown length of incisors in each group showed that first, second and third incisors (I1, I2, and I3) had the maximum to minimum crown length, respectively. The correlation between actual age and clinical crown length was strong in I1 (r = 0.73, p ≤ 0.001), I2 (r = 0.8, p ≤ 0.001), and I3 (r = 0.81, p ≤ 0.001) in the Arabian mares of Khuzestan. The correlation of estimated dental age with actual age in the Arabian mares of Khuzestan was very strong (r = 0.992, p ≤ 0.001). Therefore, the dental age estimation in the Arabian mares of Khuzestan appears to be very close to the actual age of the animal.

Quercetin improved histological structure and upregulated adiponectin and adiponectin receptors in the placenta of rats with gestational diabetes mellitus.

INTRODUCTION: Gestational diabetes mellitus (GDM) is a metabolic syndrome among pregnant mothers that increases the risk of developing growth disorders in the fetus and the placenta. Adiponectin is an adipokine, which plays a central role in the regulation of glucose and lipid metabolism, energy homeostasis, and insulin resistance in various tissues. Quercetin is a natural flavonoid with beneficial effects in the diabetic animal model, but data related to its effect on histological change and adiponectin system in the placenta of GDM are limited. In the current study, some histological changes and expression of adiponectin and its two receptors in the placenta of rats with GDM were investigated. METHODS: This study was carried out on placentas from the rodent model. To induce GDM, female rats were treated with a single dose of STZ. Placenta tissue was harvested and stained by PAS method. Protein and mRNA levels of adiponectin and its two receptors were assessed by immunohistochemistry and Real time PCR analysis, respectively. RESULTS: The results showed the increased number of glycogen cells and thickness of the labyrinth interhemal membrane (LIM) in the embryonic part of the placenta in diabetic rats, while the use of quercetin significantly prevented their increase in diabetic rats. Treatment of the diabetic group with quercetin caused significantly increased adiponectin expression and decreased its receptors.The immunohistochemical study revealed the expression of AdipoR2 in the cytoplasm of syncytiotrophoblast and cytotrophoblast cells. DISCUSSION: The results indicated that quercetin in pregnant diabetic rats could attenuate the histological abnormalities and improved adiponectin system dysregulation in the placenta.

Morphometric development of sheep (Ovis aries) lymph nodes in fetal period.

Immune system plays crucial role in body and lymph nodes are essential parts of this system for combating pathogens. However, no study has ever been conducted on morphometric development of sheep lymph nodes in fetal period. Thus, this study attempted to examine the morphometric characteristics of a number of important lymph nodes of some lymphocenters of sheep during fetal period. To this end, 60 pieces of sheep fetuses collected from Ahvaz slaughterhouse were fixated in 10% formalin and then divided into four categories based on crown-rump length (CRL) following gender and weight determinations. Mandibular, caudal superficial cervical (prescapular), caudal mediastinum, jejunal mesenteric and popliteal lymph nodes were evaluated in five lymphocenters of head, neck, thoracic cavity, abdominal viscera and pelvic limbs, respectively. In each sample, nodes formation was visually checked and in cases of nodes formation, they were measured in terms of weight, length, width and thickness and collected data were statistically analyzed. The longest and shortest fetal CRLs were found to be 48.50 cm and 3.50 cm, respectively. Gender had no effect on study parameters in 32 male and 28 female fetuses. Study of sheep fetuses' lymph nodes revealed no macroscopic lymph node development by day 45, while all nodes were observable after the day 59. The shortest lymph node was mandibular node and the longest one was caudal mediastinum. Based on the results, it seemed that although the size of lymph nodes grows by age, this increase is not the same for all nodes and groups.

Morphometric development of the vertebral segments of the vertebral column in the goat (Capra hircus) during the fetal period / Desarrollo morfométrico de los segmentos vertebrales de la columna vertebral en cabra (Capra hircus) durante el periodo fetal

Understanding normal fetal growth rates of the vertebral column, and between species, provide a basis to establish reference values for evaluation of body development and estimation of fetal age by ultrasonography. Goats are also widely used in biomedical perinatal research and are still considered a key to understanding the skeletal development. This study was carried out to clarify growth of length of vertebral column at segmental, regional, and total level of vertebral column in the fetal goats during different gestational age. The length of each vertebral segment of 25 goat fetuses, aged between 6 and 20 weeks were measured for each vertebra using a digital caliper. Our study demonstrated differences among various fetal ages in terms of regional, segmental, and total growth rate of the length of vertebral segments. With increase of fetal age, the relative length of vertebral segments of cervical, thoracic and lumbar regions diminished, whereas sacral and caudal regions increased in relative length. The thoracic vertebrae were the longest followed by cervical, lumbar, caudal, and sacral regions except at the oldest fetuses where caudal region became longer than lumbar region. Although the longest and shortest vertebral segments in cervical and lumbar regions were consistent among age groups, the trend of segmental growth of the vertebral regions was variable. Based on these detailed findings, the relative regional lengths of vertebral column were essentially different among fetal goats, humans, and neonatal rats. There is also a general trend of increasing segmental and regional initial growth and there is a relatively significant increase in growth rate caudally along the column during fetal period. This research yield important results that may be also useful for future orthopedic studies that contemplate the use of goat as a new model for the human spine.

Entender el crecimiento fetal normal de la columna vertebral entre las especies, proporciona una base para establecer valores de referencia en la evaluación del desarrollo corporal y la estimación de la edad fetal por ultrasonografía. Las cabras se utilizan frecuentemente en investigaciones biomédicas perinatales y son consideradas clave en el estudio del desarrollo esquelético. Este estudio se realizó con el objetivo de determiner el crecimiento de longitud de la columna vertebral a nivel segmentario, regional y total de la columna vertebral en cabras fetales durante diferentes etapas gestacionales. La longitud de cada segmento vertebral de 25 fetos de cabra, con edades comprendidas entre las 6 y 20 semanas se midió utilizando un calibre digital. Nuestro estudio demostró diferencias entre varias edades fetales en términos de tasa de crecimiento regional, segmentario y total de longitud de los segmentos vertebrales Con el aumento de la edad fetal, la longitud relativa de los segmentos vertebrales de las regiones cervical, torácica y lumbar disminuyó, mientras que las regiones sacras y caudales aumentaron en longitud relativa. Las vértebras torácicas fueron las más largas seguidas por las regiones cervical, lumbar, caudal y sacral excepto en los fetos más antiguos donde la región caudal se hizo más larga que la región lumbar. Aunque los segmentos vertebrales más largos y más cortos en las regiones cervical y lumbar fueron consistentes entre los grupos de edad, la tendencia de crecimiento segmentario de las regiones vertebrales fue variable. En base a estos resultados, las longitudes relativas de columna vertebral fueron esencialmente diferentes entre cabras fetales, humanos y ratas neonatas. También existe una tendencia general de aumento del crecimiento inicial segmentario y regional, como tambien un aumento relativamente significativo en la tasa de crecimiento a lo largo de la columna durante el período fetal. Esta investigación arroja importantes resultados que también pueden ser útiles para futuros estudios ortopédicos que contemplan el uso de la cabra como un nuevo modelo para la columna vertebral humana.

Effect of Enrofloxacin on Histochemistry, Immunohistochemistry and Molecular Changes in Lamb Articular Cartilage.

Enrofloxacin is a synthetic chemotherapeutic agent from the class of the fluoroquinolones that is widely used to treat bacterial infections. It is metabolized to ciprofloxacin in the body as active metabolite. Fluoroquinolones change in the articular cartilage, especially with high doses and more than two weeks use. So, due to relatively excessive use of enrofloxacin in mammals and similarity of lambs to human subjects with respect to skeletal activity cycles, this study was done to investigate the effects of enrofloxacin on some cellular and molecular changes in growing lamb articular cartilage to evaluate some possible mechanisms involved these changes. Twelve, 2-month-old male lambs divided into three groups: control group received only normal saline; therapeutic group received 5mg/kg enrofloxacin subcutaneously, daily, for 15 days and toxic group received 35 mg/kg enrofloxacin in the same manner as therapeutic group. Twenty four hours after the last dose, the animals were sacrificed, and their stifle joints were dissected. Sampling from distal femoral and proximal tibial extremities was done quickly for further histological and molecular studies. Collagen-п content was studied with avidin-biotin immunohistochemistry method in different groups. Expression of Sox9 and caspase-3 was evaluated by Real-time PCR. Immunohistochemical changes were included decreases of matrix proteoglycans, carbohydrates, and Collagen-п in the toxic group. Some of these changes were observed in the therapeutic group with less intensity in comparison to the toxic group. Enrofloxacin were significantly decreased (P≤0.05). Sox9 expression in therapeutic and toxic groups compared to control group. But caspase -3 expressions in the toxic group significantly increased (P≤0.0001) with a comparison to other groups, while, between control and therapeutic groups, there were no significant differences. So, it can be concluded that enrofloxacin increases apoptosis in chondrocytes and decreases their numbers. Enrofloxacin use in growing lambs even at recommended therapeutic dose is not completely safe on articular cartilage. Moreover, higher doses of enrofloxacin induce severe changes in lamb articular cartilage.