Experimental and Toxicologic Pathology最新文献

Sharply increasing of motor vehicles every year contributes to amounts of ultrafine particles (UFPs) in the air. Besides, the existence of UFPs in the blood may cause erythrocyte damages that subject to shape deformation. This study was aimed to investigate the influence of UFPs in the motorcycle smoke exposed to mice in different concentrations to the erythrocyte damages. The experiments were conducted by injecting the motorcycle smoke with the varied amounts in an experimental chamber (dimension of 30 × 20 × 20 cm³) where the mice were put in advance for exposuring twice a day (100 s). Total numbers of UFPs in the smoke were calculated by measuring the total concentrations multiplied by the smoke debit. They were measured using a TSI 8525 P-Trak UPC. The effects of the smoke exposures in the mice’s erythrocytes related to the UFPs in the smoke were observed by a binocular CX-31 Computer Microscope after the 2nd, 4th, 6th, 8th, and 10th exposure days. The erythrocyte damages were calculated from the total abnormal erythrocytes divided by the total erythrocytes. Our results showed that more UFPs exposed to mice resulted in more the erythrocytes damages. Longer exposures caused more damages of the mice erythrocytes. This study found significant correlations between the numbers of UFPs exposed to mice and the erythrocyte damages. Our finding gives important evidence that motorcycle emissions especially UFPs affect on health.

T. ornata a macroalgae rich in bioactive molecules possess various biological activities. Herein, the aim of the study is to evaluate the aqueous extract and the sulphated polysaccharide isolated from T. ornata for its anti-arthritic potential in Complete Freund’s Adjuvant (CFA) induced arthritis in rats. Anti-arthritic potential of aqueous T. ornata (ATO) and T. ornata sulphated polysaccharide (TSP) was evidenced by the significant reduction in paw volume and arthritic score. Inflammatory and antioxidant markers were found to be restored in the drug treated groups which was found to be in line with dexamethasone a standard anti-inflammatory drug. The histopathological and radiological examination adds on the support to the above findings confirming the anti-arthritic potential of ATO and TSP. It is interesting to note that the sulphated polysaccharide inhibits inflammation and bone damage at very low dose itself. Hence, TSP could be considered as a better candidate in the management of chronic inflammatory diseases like rheumatoid arthritis.

In the gastrointestinal tract, the immediate healing response to mucosal damage is critical to sustain mucosal homeostasis. The migration of surrounding epithelial cells to cover the denuded area without proliferation is termed restitution, followed by early reparation of the damage. In this study, we determined the role of A-kinase anchor protein 13 (AKAP13) in mice with dextran sulphate sodium (DSS)-induced colitis upon mucosal injury and restitution, and investigated whether inhibition of Rho-associated coiled-coil containing protein kinase (ROCK), downstream effector of AKAP13, affects these mucosal responses. BALB/c mice were challenged with 4% or 2% DSS in their drinking water for up to 8 or 16 days, respectively. During this period, mice received subcutaneous injections of fasudil hydrochloride hydrate (FH, 10 mg/kg, twice per day), an inhibitor of phosphorylation of ROCK. In immunohistochemistry, AKAP13 was highly expressed in the mucosal epithelium prior to DSS-induced mucosal injury, and also expressed in ulcer-covering non-proliferative epithelium, which corresponded to restituted epithelial cells. Coadministration of FH increased serum amyloid A levels and histopathological scores for mucosal injury, as compared with the DSS group. The effects were associated with a decrease in gene expression of Akap13 in the mucosal tissue and the inhibition of restitution rata (the length of restituted epithelial cells per ulcer). These results suggested that AKAP13 and ROCK are involved in mucosal response at early injury and restitution during healing in DSS-induced colitis in mice.

The liver is the primary organ participating in the metabolism of xenobiotics and is therefore an important target in the safety assessment of drugs, chemicals and environmental toxins. Drug-induced liver injury (DILI) has recently become widely recognized in human medicine as an adverse event. The progression of DILI often involves “damage-associated molecular patterns” (DAMPs) of gene and protein expression such as high-mobility group boxes (HMGBs), S100 proteins and heat shock proteins (Hsp). DAMPs are released from injured or necrotic cells and are bound to Toll-like receptors (TLRs) and modulate inflammatory reactions. Previously, in thioacetamide (TAA; 300 mg/kg body weight, single injection)-induced rat liver, we demonstrated that the expressions of DAMPs, TLR4 and major histocompatibility complex (MHC) class II were simultaneously increased, accompanied with progression of hepatocellular injury and inflammation. Here we investigated the association of DILI and DAMPs, TLRs and MHC class II by using rat livers repeated injections with TAA (100 mg/kg body weight, once, three times). Two days after TAA single injection, centrilobular hepatocellular necrosis with infiltration of mononuclear cells was observed, being paralleled with increase in serum levels of aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP). However, two days after duplicate and triplicate injections, only mild degenerative change of hepatocytes and slight infiltration of mononuclear cells were seen in the affected centrilobular area. Serum levels of AST, ALT and ALP were also decreased to the same levels of control. mRNA expressions of DAMPs (HMGBs, S100A4 and Hsp 70-2), TLR4 and MHC class II tended to be increased only on single injection, although the number of MHC class II-positive cells in the centrilobular area was still increased on each examination point. The analysis of enzymes (CYP2E1 and Flavin monooxygenase (FMO) 3), which metabolize TAA in hepatocytes, showed a significant decrease in FMO3 on the duplicate and triplicate injections. Autophagy and regulatory T cells were not significantly changed for the attenuation of hepatocyte injury. Collectively, these results suggest that hepatocytes may adapt accumulation of the toxicant by changing their enzyme functions; furthermore, MHC class II cells, which still showed increased number in the duplicate and triplicate injections, may be related with protection from the toxicant.

Phytochemicals that have antioxidant effect play important role in protection against several diseases in humans. This study was carried out to evaluate the efficacy of hesperidin and tiger nut against the early changes that may be related to the toxicity of acrylamide in female rats. 72 Sprague Dawley female rats were divided into six groups (12 rat/group): control group (I); hesperidin (HES) treated group (II); tiger nut (TN) treated group (III); Acrylamide (ACR) treated group (IV); HES-ACR treated group (V); and TN-ACR treated group (VI). There was a significant increase in the levels of serum carcino embryonic antigen (CEA), malondialdehyde (MDA), protein carbonyls (CO), ALT, AST, LDH, urea and creatinine while no significant changes of serum total sialic acid, progesterone (prog) and estradiol (E2) levels, and significant decreases of body weights, catalase (Cat) activity, superoxide dismutase (SOD) activity, reduced glutathione (GSH) level, and glutathione peroxidase (GSH-Px) activity of ACR treated group compared with the control. Our results suggested that supplementation of a diet with hesperidin provided antioxidant defense more significant than tiger nut against the toxicity of ACR in breast, liver and kidney tissues.

Deoxynivalenol (DON) and zearalenone (ZEA) are the two most common mycotoxins in animal feed. In this study, we examined oxidative injury and apoptosis of porcine splenic lymphocytes induced by DON or ZEA and their combination in vitro. Based on IC₅₀ values, porcine splenic lymphocytes were treated with 0.06, 0.3, 1.5, and 7.5 μg/mL DON, 0.08, 0.4, 2, and 10 μg/mL ZEA, or both DON and ZEA at 0.06 and 0.08 μg/mL, 0.3 and 0.4 μg/mL, and 1.5 and 2 μg/mL, respectively. After 48 h of DON and/or ZEA exposure, the cells were analyzed for antioxidant functions, apoptosis, and mRNA and protein expression of apoptosis-related genes p53, Bcl-2, Bax, caspase-3, and caspase-8 to determine their apoptosis and oxidative damage effects and mechanisms. The results showed that, compared with the control group, SOD, CAT, GPx, GSH, and Bcl-2 mRNA and protein expression levels were significantly reduced in exposed groups (P < 0.05 or P < 0.01). Furthermore, MDA contents, apoptosis rates, and p53, Bax, caspase-3 and caspase-8 protein and mRNA expression levels were increased significantly (P < 0.01). The effects of DON and ZEA were dose dependent and synergistic in combination. These data suggest that DON and ZEA induce oxidative damage and apoptosis of porcine splenic lymphocytes.

The study was conducted to investigate the protective role of α-tocopherol against polychlorinated biphenyls (PCBs) induced effect in Sertoli cell function of F₁ prepuberal rats. Dams were grouped into six; each group consists of six animals. Group 1–control treated with corn oil as vehicle, group II- 0.5 mgPCBs/kg bw/day, group III- 0.5mgPCBs/kg bw/day with α-tocopherol (50 mg α-tocopherol/kg bw/day), group IV- 1mgPCBs/kg bwt/day, group V- 1mgPCBs/kg bw/day with α-tocopherol (50 mg α-tocopherol/kg bw/day) and group VI − α-tocopherol alone treated orally from postpartum day1-20. Male offspring rats were euthanized on post natal day 21. Testes were collected for the histological examination and Sertoli cell isolation. The protein levels of follicle-stimulating hormone receptor, androgen binding protein, androgen receptor, estrogen receptor α & β, Inhibin-β, transferrin, claudin-11, occludin, E-cadherin, connexin-43, c-fos, c-jun, SF1, USF1 & 2 were studied using western blot method. The testicular architecture was affected in the PCBs exposed rats but this effect was restored by α-tocopherol supplementation. PCBs decreased the protein levels of FSHR, AR, ABP, ERα & β, transferrin, claudin-11, occludin, E-cadherin, connexin-43, c-fos, c-jun, SF1, USF1 & 2 whereas inhibin-β protein level was found to be increased in Sertoli cells. These results suggested that α-tocopherol has ameliorative role against PCBs induced testicular Sertoli cell dysfunction in F₁ progeny.

We aimed to explore the cytotoxic and apoptotic effect of friedelin on breast cancer MCF-7 cells. Cytotoxic effect of friedelin on MCF-7 cells was analyzed using MTT, cell and nuclear morphology. The apoptosis mechanism of friedelin on MCF-7 cells was analyzed using real-time PCR. Friedelin potentially inhibit 78% of MCF-7 cell’s growth, the IC₅₀ value was 1.8 μM in 24 h and 1.2 μM in 48 h. Friedelin increased ROS significantly and DNA damage was confirmed by tunel assay. We found characteristically 52% apoptotic cells and 6% necrotic cells in PI, AO/ErBr staining after 48 h treatment with 1.2 μM of friedelin. Apoptosis was confirmed by significantly (p ≤ 0.001) increased tumor suppressor gene Cdkn1a, pRb2, p53, Nrf2, caspase-3 and decreased Bcl-2, mdm2 & PCNA expression after 48 h. In conclusion, friedelin effectively inhibit breast cancer MCF-7 cell growth, it was associated with early expression of Cdkn1a, pRb2 and activation of p53 and caspases.

The present study was aimed to investigate the effect of triiodothyronine (T3) on the improvement of articular cartilage surface architecture at in vitro level. The T3 hormone was applied to neo-tissues in the range of 50, 100, 150 and 200 ng/ml for 5 weeks. At the end of the treatment, biochemical and histological evaluation was carried out in the neo-tissues. T3 hormone application significantly increased the collagen production in neo-cartilage tissues. The properties of tensile and compressive were significantly increased compared to the controls. However, T3 hormone application also induced hypertrophy. At the higher dose concentration of T3 hormone application, tensile and compressive properties were tremendously increased 4.3 and 4.6 fold respectively. Taking all these data together, it suggested that the T3 hormone application could be a potential agent to increase the functional properties such tensile and compressive in neo-tissues.

There are two types of bronchiolo-alveolar hyperplasia (hyperplasia) in rodent lungs. The first is “inflammatory hyperplasia” that retains its ability to revert to normal epithelia upon removal of the stimulating insult. The second is “latent tumorigenic hyperplasia”, which is irreversible and causes independent preneoplastic lesions that can progress to bronchiolo-alveolar adenocarcinoma. Previously, lung samples with hyperplastic lesions were obtained from rats exposed to N-bis (2-hydroxypropyl) nitrosamine (DHPN) and fine particles (e.g. quartz), and 19 specific markers were examined immunohistochemically to identify latent tumorigenic hyperplasia. In the cytoplasm of the cells that make up the alveolar wall, we found that napsin A was weakly expressed in the inflammatory hyperplastic lesions, and was strongly expressed in the latent tumorigenic hyperplastic lesions induced by DHPN.

To validate the possibility that napsin A may serve as a tumorigenic hyperplastic marker, additional experiments were performed with rats and mice. Latent tumorigenic hyperplasia induced by various carcinogens were positive for napsin A, similar to hyperplasia induced by DHPN. Thus, high expression of napsin A in alveolar walls may serve as a useful marker for detecting the tumorigenic potential of lung hyperplasia in rodents.