ACR-treated Group Research Articles

Fagopyrum tataricum (L.) Gaertn interacts with Gsk-3β/Nrf-2 signalling to protect neurotoxicity in a zebrafish model

Ethnopharmacological relevanceFagopyrum tataricum (L.) Gaertn is used as a folk medicine in many Asian countries due to its anti-inflammatory, antioxidant, and several other health-promoting properties. It is also prescribed to improve neurocognitive functions and alleviate inflammatory conditions. Aim of the studyOxidative stress and neuroinflammation plays a crucial role in neurodegenerative conditions. Hence, based on the ethnomedical claims and available literature, the present study investigated neuroprotective efficacy of a seed extract (ft-ext) of Fagopyrum tataricum against acrylamide (ACR)-induced neurotoxicity. Materials and methodsThe phytochemical characterization of ft-ext was performed by a high-performance liquid chromatography method. Molecular interactions of the identified compounds of ft-ext were studied using an in-silico docking tool. An in-vitro protein denaturation assay was done to check anti-inflammatory activity. The 5 days’ post-fertilized zebrafish larvae were exposed to 1 mM and 2.5 mM ACR with or without ft-ext for 72 h to study its neuroprotective efficacy. Real-time polymerase chain reaction and western blotting studies were performed to analyse the oxidative stress-related gene and protein expressions respectively. ResultsThe extract showed the presence of chlorogenic acid, rutin, caffeic acid, vitexin, syringic acid, quercetin, p-coumaric acid, kaempferol, and ferulic acid. In-vitro protein denaturation assay of ft-ext showed a potent anti-inflammatory effect. The ft-ext improved ACR-mediated locomotor deficit and reduced overall mortality in the larvae. The brain lipid peroxidation and protein carbonylation results revealed an elevated level of oxidative stress in the ACR-treated group, which was reduced in ft-ext-treated larvae. The extract treatment increased the expression of nrf2, gpx, and hmox1a, while simultaneously downregulated trxr2 levels in the brain of larvae exposed to ACR. The treatment also showed inactivation of Gsk-3β, thus maintaining a normal pool of Nrf2 and β-catenin. Molecular docking of identified compounds of ft-ext showed possible hydrogen and hydrophobic interactions with Gsk-3β. ConclusionThe ft-ext prevents ACR-mediated neurotoxicity by suppressing Gsk-3β mediated oxidative stress.

Effect of vitamin C on histomorphometric changes of testis, epididymis, prostate, and seminal vesicle induced by acrylamide in rat.

Acrylamide (ACR) has adverse effects on the rat testis. This study aimed to assess the impact of ACR and vit C exposure on reproductive organs in rats. In this experimental study, 32 adult male rats were used. The animals were divided into 4 groups (n = 8): (1) control group, (2) ACR (10mg/kg) group, (3) vit C (200mg/kg), (4) ACR (10mg/kg) + vit C (200mg/kg) daily for 5weeks by gavage. After the administration period, testis, prostate, seminal vesicle, and epididymis of animals are removed; after preparing tissue sections, the structural changes of the tissues are examined by stereology. Data were analyzed using one-way ANOVA followed by the Tukey test in SPSS software. A value of p ≤ 0.05 was considered significant. The testis weight, volume (mm3), and the mean Johnsen score showed a significant decrease in comparison with the control group and vit C-treated group. These parameters were increased in ACR + vit C group. The number of spermatogonia, spermatocyte, spermatid, and Sertoli and Leydig cells in ACR-treated group showed a significant decrease in comparison with the control and vit C-treated groups. The number of these cells was increased in the ACR + vit C group. Epithelium height and folding of prostate and seminal vesicle in the ACR-treated group were decreased. Epithelium lost its integrity. In the ACR + vit C group, histopathological changes were decreased. Seminal vesicle of ACR + vit C-treated group showed mild degeneration and rupture in epithelium integrity. The epididymis of ACR + vit C group also showed mild degeneration and rupture in epithelium integrity.

The abrogative effect of propolis on acrylamide-induced toxicity in male albino rats: Histological study

Abstract The present study investigated the toxic action of acrylamide (ACR) and the abrogative effect of aqueous propolis extract on ACR-induced toxicity in male albino rats. Forty adult male albino rats were randomly divided into four groups and administered the treatment doses orally by gavage. Control, treated with a physiological solution (5.0 ml/rat). ACR-treated group, treated with ACR 25 mg/kg. ACR + propolis, treated with 25 mg/kg ACR + 100 mg/kg propolis. Propolis-treated group, treated with 100 mg/kg. The treatment period was 28 days, the treatment doses were administered orally using a gavage stomach tube. The results showed that the presence of ACR increased (P < 0.001) the level of liver enzymes alanine aminotransferase (167.2 vs 90.15) and aspartate aminotransferase (120.66 vs 41.52) in the liver tissue serum and lipid peroxidation products (80.11 vs 39.3); also, it decreased (P < 0.001) the total capacity of antioxidants (113.4 vs 189.41) compared to the control group. The histological analysis of the kidney revealed alterations induced by ACR, including atrophy, necrosis, renal glomeruli atrophy, tubular necrosis, enlargement of the glomeruli, hemorrhage, and edema surrounding the blood vessels. Also, the results showed that the rats treated with propolis improved liver and kidney functions because of propolis’s efficiency against the harmful effects of ACR. Moreover, the histological examination of the liver tissue revealed negative changes, with focal necrosis in hepatocytes. Microscopic examination showed tubular necrosis within the seminiferous tubules, sloughing, and desquamation of sperm cells into the lumen. Also, an infiltration of protein substances between the tubules and degenerative vacuolar changes between sperm cells was observed. The renal, hepatic, and testicular tissues appeared almost similar to the control group, except for some minor changes. This study proved that feeding the rats with propolis reduced these pathological effects and restored the tissues to a normal state. It is concluded that using propolis with ACR significantly reduced the biochemical and histological damage caused by ACR, and recommends using propolis as a cytoprotective agent against pathological toxicity of ACR.

Prenatal exposure to acrylamide differently affected the sex ratio, aromatase and apoptosis in female adult offspring of two subsequent generations.

In the present study, we investigated the effect of acrylamide (ACR) exposure during pregnancy on the ovary of female adult offspring of two subsequent generations. Sixty-day-old Wistar albino female rats were given different doses of ACR (2.5 and 10 mg/kg/day) from day 6 of pregnancy until giving birth. Females from the first generation (AF1) were fed ad libitum, and thereafter, a subgroup was euthanized at 8 weeks of age and ovary samples were obtained. The remaining females were maintained until they reached sexual maturity (50 days old) and then treated in the same way as the previous generation to obtain the second generation of females (AF2). The histopathological examination indicated a high frequency of corpora lutea along with an increased number of antral follicles that reached the selectable stage mainly at a dose of 2.5 mg/kg/day. Interestingly, ACR exposure significantly increased the mRNA levels of CYP19 gene and its corresponding CYP19 protein expression in AF1 females. The TUNEL assay showed a significantly high rate of apoptosis in stromal cells except for dose of 2.5 mg/kg/day. However, in AF2 females, ACR exposure significantly increased the number of degenerating follicles and cysts while the number of growing follicles was reduced. Moreover, in both ACR-treated groups, estradiol-producing enzyme CYP19A gene and its corresponding protein were significantly reduced, and an excessive apoptosis was produced. We concluded that the ovarian condition of AF1 females had considerable similarity to the typical early perimenopausal stage, whereas that of AF2 females was similar to the late perimenopausal stage in women.

Fetal programming: in utero exposure to acrylamide leads to intergenerational disrupted ovarian function and accelerated ovarian aging.

In this study we investigated the effects of multigenerational exposures to acrylamide (ACR) on ovarian function. Fifty-day-old Wistar albino female rats were divided into the control and ACR-treated groups (2.5, 10, and 20 mg/kg/day) from day 6 of pregnancy until delivery. The obtained females of the first (AF1) and second generation (AF2) were euthanized at 4 weeks of age, and plasma and ovary samples were collected. We found that in utero multigenerational exposure to ACR reduced fertility and ovarian function in AF1 through inducing histopathological changes as evidenced by the appearance of cysts and degenerating follicles, oocyte vacuolization, and pyknosis in granulosa cells. TMR red positive cells confirmed by TUNEL assay were mostly detected in the stroma of the treated groups. Estradiol and IGF-1 concentrations significantly decreased as a result of decreased CYP19 gene and its protein expression. However, ACR exposure in AF2 led to early ovarian aging as evidenced by high estradiol and progesterone levels among all treated groups compared to control group, corresponding to the upregulation of the CYP19 gene and protein expression. The apoptotic cells of the stroma were greatly detected compared to that in the control group, whereas no significant difference was reported in ESR1 and ESR2 gene expression. This study confirms the developmental adverse effects of ACR on ovarian function and fertility in at least two consecutive generations. It emphasizes the need for more effective strategies during pregnancy, such as eating healthy foods and avoiding consumption of ACR-rich products, including fried foods and coffee.

Reproductive Injury in Male Rats from Acrylamide Toxicity and Potential Protection by Earthworm Methanolic Extract.

This study examined the protective effect of earthworm extract (EE) on acrylamide (ACR)-induced reproductive dysfunction. Forty male rats were allocated into four groups (n = 10). The G I (control) group received distilled water (D.W.). The G II group received ACR (5 mg kg-1 B.W. in D.W.) 5 days per week, orally, for 3 weeks. The G III group was administered EE (300 mg kg-1 B.W in D.W.) 5 days per week, orally, for 3 weeks. The G IV group was pretreated with EE for 3 weeks and then co-treated with EE and ACR for an additional 3 weeks. ACR decreased the number of sperm, sperm viability, and total motility. However, it increased testosterone levels with no effect on the FSH or LH levels. Moreover, ACR increased the concentrations of malondialdehyde (MDA) and nitric oxide (NO). Meanwhile, it decreased the glutathione (GSH) concentration in testicular tissues. Notably, the expression levels of p53 and Ki-67 were increased in the degenerated spermatogenic cells and in the hyperplastic Leydig cells of the testis of the ACR-treated group, respectively. Acrylamide induced alterations in the testicular tissue architecture. Interestingly, EE restored the sperm parameters and recovered the testicular histological structures and the biochemical alterations induced by ACR. In conclusion, earthworm extract ameliorated ACR-induced reproductive toxicity via restoring the testicular antioxidant balance and suppressing p53 and Ki-67 expressions in testicular tissues.

AQP4 Attenuated TRAF6/NFκB Activation in Acrylamide-Induced Neurotoxicity.

Acrylamide (ACR) is present in high-temperature-processed high-carbohydrate foods, cigarette smoke, and industrial pollution. Chronic exposure to ACR may induce neurotoxicity from reactive oxygen species (ROS); however, the mechanisms underlying ACR-induced neurotoxicity remain unclear. We studied 28-day subacute ACR toxicity by repeatedly feeding ACR (0, 15, or 30 mg/kg) to rats. We conducted RNA sequencing and Western blot analyses to identify differences in mRNA expression in the blood and in protein expression in the brain tissues, respectively, of the rats. AQP4 transient transfection was performed to identify potential associations with protein regulation. The rats treated with 30 mg/kg ACR exhibited hind-limb muscle weakness. Matrix metalloproteinase (MMP9) expression was higher in the ACR-treated group than in the control group. ACR induced MMP-9 and AQP4 protein expression in the brain tissues of the rats, which subsequently presented with neurotoxicity. In the in vitro study, Neuro-2a cells were transiently transfected with AQP4, which inhibited MMP-9 and TNF receptor-associated factor 6 (TRAF6) expression, and inhibited ACR induced expression of TRAF6, IκBα, and nuclear factor κB (NFκB). Using a combination of in vivo and in vitro experiments, this study revealed that depressive symptoms associated with ACR-induced neurotoxicity are associated with downregulation of AQP4 and induction of the TRAF6 pathway.

Ameliorative activity of medicinal plant fraction for neuroprotection against acrylamide-induced neurotoxicity in Drosophila melanogaster\u2014a comparative study

BackgroundMedicinal plant extracts used in folk medicine seem to be crucial since those are generally accepted by people without worrying about the toxicity. In our present study, we have compared the neuroprotective role of the rhizome of Curculigo orchioides Gaertn. and leaf extracts of Olea dioica Roxb., against acrylamide-induced neurotoxicity in Drosophila melanogaster.ResultsIn-vivo neurotoxic study was carried out using 7-day-old flies (wild-type D. melanogaster). Prior to co-after exposing the flies with acrylamide (8 mM) along with or without OLE-2 and CU-3 fractions (0.2, 0.4 and 0.6% w/v), the heads of flies of both the control and treated groups were homogenized for biochemical assay. ACR-treated groups have shown higher elevation in AChE, SOD, LPO, and CAT activity when compared to control and treated (plant fraction) groups. Biochemical and histopathology studies show that both the plant fractions (OLE-2 and CU-3) have neuroprotective action against acrylamide.ConclusionThe present study has demonstrated that dietary supplementation with plant fraction of OLE-2 and CU-3 has increased antioxidant enzymes and decreased AChE activity in D. melanogaster. This activity could be part of the probable mechanisms of action by which OLE-2 and CU-3 fractions have extended the lifespan and locomotory activity in fruit flies. These fruit flies continue to provide an exciting system for metabolic studies and should be more extensively exploited. Based on these results, further studies on the identified bioactive compounds from these two plants are being planned.

Exposure to acrylamide inhibits uterine decidualization via suppression of cyclin D3/p21 and apoptosis in mice

Acrylamide (ACR), a neurotoxicity and carcinogenic chemical, has attracted considerable attention since it is present at high concentrations in thermally cooked carbohydrate-rich foods. ACR exposure significantly increased rate of fetal resorption, and decreased fetal body weights in mice. However, no detailed information is available about the effect of ACR on uterine decidualization, which is a vital process in the establishment of successful pregnancy. Thus, our aim of this study was to explore the effect and mechanism of ACR on uterine decidualization in vivo during mice pregnancy. Mice were gavaged with 0, 10, and 50 mg ACR /kg/day from gestational days (GD) 1 until GD 8, whereas pseudopregnant mice from pseudopregnant day (PPD) 4 until PPD 8. Results indicated ACR treatment dramatically reduced numbers of implanted embryos, and decreased the weights of implantation site and oil-induced uterus. Nevertheless, no significant difference was observed in the weights of no oil-induced uterus between control and ACR-treated group. Furthermore, ACR significantly reduced numbers of polyploidy and PCNA-positive decidual cells and expression of cyclin D3 and p21 proteins, and induced apoptosis of decidua, as presented by up-regulation of Bax and cleaved-caspase-3, and decreased Bcl-2 protein during normal pregnant and pseudopregnant process. In summary, ACR exposure significantly inhibited uterine endometrial decidualization via the apoptosis and suppression of cyclin D3/p21 in mice.

Redox status and fatty acid composition of Mactra corallina digestive gland following exposure to acrylamide.

Acrylamide (ACR), a ubiquitous agent, has various chemical and industrial applications, and it is found in backed or fried carbohydrate-rich food. It has been related to multiple toxicological effects, and it causes high cytotoxicity through oxidative stress. The present study aimed to investigate the potential effect of ACR toxicity administered at different concentrations (5, 10, and 20mg/L), during 5days, in order to evaluate the fatty acid (FA) composition and redox state in the digestive gland of Mactra corallina. The results showed, in ACR-treated clams, a significant increase in malondialdehyde, hydrogen peroxide, protein carbonyl, and metallothionein levels, as well as an alteration of the enzymatic (superoxide dismutase, glutathione peroxidase, and catalase) and non-enzymatic (reduced glutathione and ascorbic acid) antioxidant status. However, acetylcholinesterase activity was inhibited in a concentration-dependent manner. In our experiment, the n-3 (Omega-3) and n-6 (Omega-6) polyunsaturated fatty acid levels were significantly changed in all ACR-treated groups. A decrease in eicosapentaenoic acid (C20:5n-3, EPA) and docosahexaenoic acid (C22:6n-3, DHA) was observed in 10-mg/L and 20-mg/L ACR-treated groups. Nevertheless, arachidonic acid (C20:4n-6, ARA) and its precursor linoleic acid (C18:2n-6, LA) were increased. Besides oxidative stress parameters, FA composition may be an additional tool for assessing ACR contamination.

Evaluation of epigallocatechin gallate and epicatechin gallate effects on acrylamide-induced neurotoxicity in rats and cytotoxicity in PC 12 cells

The neurotoxicity of acrylamide (ACR) monomer occurs through different mechanisms such as oxidative stress. Epigallocatechin gallate (EGCG) and epicatechin gallate (ECG) are green tea catechins which are known as powerful antioxidants. In this study, we examined the possible protective effects of ECG and EGCG on ACR neurotoxicity in both in-vitro and in-vivo models. PC12 cells were exposed to different concentrations of ECG and EGCG. After 24 and 48 hours, ACR was added to the cells (IC50 = 4.85 mM) and cell viability was measured through MTT assay after 24 hours. Male Wistar rats were pretreated with ECG, EGCG (10, 20 and 40 mg/kg, i.p) and vitamin E (200 IU/kg i.p.) for 3 days. Afterwards they were treated with ACR (50 mg/kg, i.p.) for 11 days. After the treatment period, gait score examination was performed and molondialdehyde (MDA) and reduced glutathione (GSH) were measured in cerebral cortex. ACR reduced the cell viability in a concentration-dependent manner. Both ECG and EGCG (20 μM) showed inhibitory effects on ACR cytotoxicity. ACR significantly induced gait abnormalities, decreased GSH level and increased lipid peroxidation in cerebral cortex. ECG and EGCG (20 mg/kg) improved all ACR toxic effects. Although the food intake was increased in pretreated groups compared to the ACR-treated group, intensive weight loss was observed due to the green tea’s different weight loss mechanisms. ECG and EGCG inhibited the cytotoxicity of ACR in PC12 cells and increased GSH level and decreased lipid peroxidation in rat cerebral cortex.

Metformin Protects against Experimental Acrylamide Neuropathy in Rats.

Preclinical Research To investigate the potential neuroprotective effects of metformin against experimental acrylamide neuropathy in rats, 24 rats were distributed into four equal groups (6 each). Group 1 was kept as a control. Group 2 (MET) was orally given metformin (200 mg/kg BW/day). Group 3 (ACR) was injected IP with acrylamide (50 mg/kg BW/day). Animals in group 4 (ACR + MET) were administered both MET and ACR at the same dose and route used in groups 2 and 3. Treatments were administered three times a week for three weeks. ACR induced an increase in lipid peroxidation in brain and spinal cord. This was associated with down regulation of bcl2 and up regulation of caspase3 in cerebrum, cerebellum, spinal cord, and sciatic nerve in the ACR-treated group. ACR-treated rats revealed neuronal degeneration and glial cell reaction in brain and spinal cord with axonal degeneration and myelin sheath irregularities in sciatic nerve. MET restored lipid peroxidation in brain and spinal cord, decreased caspase3 activity and up regulated bcl2 expression in cerebrum and sciatic nerve. Histopathological findings in ACR + MET group were lesser severe than those established in ACR-group indicating that MET ameliorates the neuropathic effects of ACR in rats. Drug Dev Res 78 : 349-359, 2017. © 2017 Wiley Periodicals, Inc.

Acrylamide-induced Subacute Neurotoxic Effects on the Cerebral Cortex and Cerebellum at the Synapse Level in Rats

ObjectiveTo investigate acrylamide (ACR)-induced subacute neurotoxic effects on the central nervous system (CNS) at the synapse level in rats. MethodsThirty-six Sprague Dawley (SD) rats were randomized into three groups, (1) a 30 mg/kg ACR-treated group, (2) a 50 mg/kg ACR-treated group, and (3) a normal saline (NS)-treated control group. Body weight and neurological changes were recorded each day. At the end of the test, cerebral cortex and cerebellum tissues were harvested and viewed using light and electron microscopy. Additionally, the expression of Synapsin I and P-Synapsin I in the cerebral cortex and cerebellum were investigated. ResultsThe 50 mg/kg ACR-treated rats showed a significant reduction in body weight compared with untreated individuals (P < 0.05). Rats exposed to ACR showed a significant increase in gait scores compared with the NS control group (P < 0.05). Histological examination indicated neuronal structural damage in the 50 mg/kg ACR treatment group. The active zone distance (AZD) and the nearest neighbor distance (NND) of synaptic vesicles in the cerebral cortex and cerebellum were increased in both the 30 mg/kg and 50 mg/kg ACR treatment groups. The ratio of the distribution of synaptic vesicles in the readily releasable pool (RRP) was decreased. Furthermore, the expression levels of Synapsin I and P-Synapsin I in the cerebral cortex and cerebellum were decreased in both the 30 mg/kg and 50 mg/kg ACR treatment groups. ConclusionSubacute ACR exposure contributes to neuropathy in the rat CNS. Functional damage of synaptic proteins and vesicles may be a mechanism of ACR neurotoxicity.

In Vivo acrylamide exposure may cause severe toxicity to mouse oocytes through its metabolite glycidamide.

High acrylamide (ACR) content in heat-processed carbohydrate-rich foods, as well as roasted products such as coffee, almonds etc., has been found to be as a risk factor for carcinogenicity and genotoxicity by The World Health Organization. Glycidamide (GLY), the epoxide metabolite of ACR, is processed by the cytochrome P-450 enzyme system and has also been found to be a genotoxic agent. The aim of this study was to determine whether ACR and/or GLY have any detrimental effect on the meiotic cell division of oocytes. For this purpose, germinal vesicle-stage mouse oocytes were treated with 0, 100, 500, or 1000 μM ACR or 0, 25, or 250 μM GLY in vitro. In vivo experiments were performed after an intraperitoneal injection of 25 mg/kg/day ACR of female BALB/c mice for 7 days. The majority of in vitro ACR-treated oocytes reached the metaphase-II stage following 18 hours of incubation, which was not significantly different from the control group. Maturation of the oocytes derived from in vivo ACR-treated mice was impaired significantly. Oocytes, reaching the M-II stage in the in vivo ACR-treated group, were characterized by a decrease in meiotic spindle mass and an increase in chromosomal disruption. In vitro GLY treatment resulted in the degeneration of all oocytes, indicating that ACR toxicity on female germ cells may occur through its metabolite, GLY. Thus, ACR exposure must be considered, together with its metabolite GLY, when female fertility is concerned.

The effect of lipoic acid on acrylamide-induced neuropathy in rats with reference to biochemical, hematological, and behavioral alterations

Context: Acrylamide (ACR) is a well-known neurotoxicant and carcinogenic agent which poses a greater risk for human and animal health.Objective: The present study evaluates the beneficial effects of α-lipoic acid (LA) on ACR-induced neuropathy.Materials and methods: A total of 40 male rats were divided into four groups: a placebo group; LA-treated group, administered orally 1% (w/w) LA mixed with diet; ACR-treated group, given 0.05% (w/v) ACR dissolved in drinking water; and LA + ACR-treated group, given LA 1% 7 d before and along with ACR 0.05% for 21 d. After 28 d, blood samples were collected, the rats were decapitated, and the tissues were excised for the measurement of brain biomarkers, antioxidant status, and hematological analysis. Also, the gait score of rats was evaluated.Results: ACR-exposed rats exhibited abnormal gait deficits with significant (p < 0.05) decline in acetylcholine esterase (AChE) and creatine kinase in serum and brain tissues, respectively. However, the lactate dehydrogenase activity was increased in serum by 123%, although it decreased in brain tissues by −74%. ACR significantly (p < 0.05) increased the malondialdehyde level by 273% with subsequent depletion of glutathione S-transferase (GST), glutathione peroxidase (GPx), and glutathione reductase (GR) activities and reduced the glutathione (GSH) level in brain tissue. Interestingly, LA significantly (p < 0.05) improved brain enzymatic biomarkers, attenuated lipid peroxidation (LPO), and increased antioxidant activities compared with the ACR-treated group.Discussion and conclusion: These results suggested that LA may have a role in the management of ACR-induced oxidative stress in brain tissues through its antioxidant activity, attenuation of LPO, and improvement of brain biomarkers.

Immunotoxicity of Acrylamide in Female BALB/c Mice

ObjectiveTo investigate the immunotoxicity of acrylamide (ACR) in female BALB/c mice. MethodsA total of 200 female mice weighing 18–22 g were randomly divided into four clusters based on body weight, and each weight-based cluster included five groups (10 mice per group): negative control, positive control (cyclophosphamide), low, intermediate, and high dose ACR groups, and all the groups were administered ACR by gavage for 30 days. At the end of the study, the immunotoxicological effects of the ACR were evaluated through immunopathology, humoral immunity, cellular immunity, and non-specific immunity. ResultsThe terminal body weight, spleen and thymus weights, lymphocyte counts in the ACR-H group were decreased, pathological changes were observed in lymph glands, thymus and spleen. %T cells in blood lymphocytes were significantly increased in all ACR-treated groups, and a significant reduction of % natural killer(NK) cells and increase of %Th cells were observed in the ACR-H group. interleukin-6(IL-6), Concanavalin A(ConA)-induced splenocyte proliferation and serum half hemolysis value (HC50) were also significantly suppressed in the ACR-H group. ConclusionACR elicited an inhibitory effect on cellular and humoral immunity of mice after 30 day feeding.

Responses of acrylamide-treated rat bladders

Acrylamide (ACR) is a chemical used in many industries around the world and more recently was found to be formed naturally in foods cooked at high temperatures. ACR was shown to be a neurotoxicant, reproductive toxicant, and carcinogen in animal species. The aim of the present study is to evaluate the influence of ACR treatment on urinary bladder responses to carbachol (10-9-3x10-4 M) and potassium chloride (KCl; 5-100 mM), each of them causes receptor-dependent and receptor-independent contractions, respectively. We also examined the role of gender in these responses. Rats of both genders were divided into three groups as follows: (1) Control animals (2) ACR-I; ACR-treated (2 mg/kg-d for 90 days) (3) ACR-II; ACR-treated (5 mg/kg-d for 90 days). In rats treated with ACR, the EC50 values of carbachol and KCl, but not the maximal response, to both agents were significantly higher than in control group. Histopathological parameters such as edema, congestion, inflammatory cells, microvascular proliferation, fibrosis, eosinophils, mast cells and epithelial damage were all higher in the ACR-treated group than in the controls. These results demonstrate for the first time that ACR-treatment can induce urinary bladder injury (Tab. 4, Fig. 4, Ref. 30).

The cyclophosphamide metabolite, acrolein, induces cytoskeletal changes and oxidative stress in Sertoli cells

The aim of this study is to explore the mechanism by which acrolein (ACR), a metabolite of cyclophosphamide (CP), induces immature Sertoli cell cytoskeletal changes. Sertoli cells obtained from rats were cultivated and treated with 50 and 100μM ACR. XTT assays were performed to detect cell viability. Activities of superoxide dismutase (SOD), glutathione peroxidases (GSH-Px), and catalase (CAT), as well as total anti-oxidation competence (T-AOC) were examined. Superoxide anion levels were detected by a fluorescent probe. Cell ultrastructure changes were observed by transmission fluorescent microscope. Actin filament (F-actin) distribution was detected by immunofluorescence, and ERK and p38MAPK expression were detected by western blot analysis. ACR significantly decreased the viability of Sertoli cells in a dose- and time-dependent manner. T-AOC and the antioxidant activity of SOD, CAT and GSH-Px, were decreased in ACR-treated groups compared with the control group. The levels of reactive oxygen species (ROS) in ACR-treated Sertoli cells were increased. In addition, characteristics of cell apoptosis such as mitochondrial swelling, aggregated chromatin, condensed cytoplasm, nuclei splitting, and nuclei vacuolization were observed in ACR-treated cells. Furthermore, ACR-treatment also induced microfilament aggregation, marginalization and regionalization. The expression levels of ERK and p38MAPK were also increased in ACR-treated cells in a dose- and time-dependent manner. ACR, a major CP metabolite, impairs the cytoskeleton which is likely caused by induction of the oxidative stress response through up-regulation of ERK and p38MAPK expression.

Chemoprevention of Barley and Sage against acrylamide-Induced genotoxic, biochemical and histopathological alterations in rats

Acrylamide (ACR) has recently been found in fried and backed foods, suggesting widespread public exposure. ACR is an industrial chemical material that causes neurotoxicity in humans and was designated as a probable human carcinogen by IARC and USEPA. The aim of the present study was to evaluate the protective effects of barley and sage against ACR mutagenicity and biochemical and histopathological changes in rats. Forty mature male rats were divided into eight groups and were fed barley and/or sage-supplemented diet (5%) with or without ACR (50 mg/kg b.w). The biochemical results revealed that ACR increased Alanine amino transferase (ALT), Aspartate amino transferase (AST), triglycerides (TG), cholesterol and uric acid. Micronucleated polychromatic erythrocytes and chromosomal aberrations in somatic and germ cells were significantly increased in ACR-treated animals. Severe pathological lesions included testicular degeneration, oedema, spermatid giant cells formation and necrosis of spermatid cells were found in the testis of ACR-treated group. The kidney of this group showed degenerative changes. Cotreatment with barley and/or Sage and ACR resulted in a significant improvement in all the parameters tested. It could be concluded that these plants contain antioxidant compounds and may be useful when add as food additive to the food cooked on a higher temperature.

Comparative Study of Modification and Degradation of Neurofilament Proteins in Rats Subchronically Treated with Allyl Chloride, Acrylamide, or 2,5-Hexanedione

Allyl chloride (ALL), acrylamide (ACR), and 2,5-hexanedione (2,5-HD) are all industrial neurotoxicants and known to produce accumulation of neurofilament (NF) proteins in both the central and peripheral nervous systems. To clarify whether any common mechanisms underlie these neurofilamentous axonopathies, the ability of ALL, ACR, and 2,5-HD to cross-link the NFs and the effects on NF degradation by Ca2+-activated neutral protease were investigated in spinal cords from rats subchronically treated with these chemicals. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis followed by immunoblot analysis revealed the appearance of high-molecular-weight species of NF triplets immunoreactive to each anti-68K, anti-160K, and anti-200K NF antibody in the 2,5-HD-treated rats, whereas it was not found in those treated with ALL or ACR. A time course study on the degradation of NF proteins conducted by the co-incubation with Ca2+ showed degradation resistance in all three NF subunits from animals treated with 2,5-HD, while no significant alterations in the rate of NF degradation were observed in the ALL- or ACR-treated group. The present results suggest that neurofilament-filled axonopathy induced by ALL or ACR and axonopathy induced by 2,5-HD may not share a common mechanism, though the initial step for the pathogenesis of this chemically induced neurotoxicity is not fully understood at present.