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Evaluation of sperm quality in mice exposed to camphor and protective role of Vitamin E
2016
adibmoradi, masood | kalantari hesari, ali | morovvati, Hassan | asadi, farzad | Moradi, Hamid Reza
BACKGROUND: In traditional medicine in some Asian countries, including Iran, there is a belief that camphor is a suppressor of sexual activity. Not only has the validity of this hypothesis not been established, but also studies in this field are very limited. OBJECTIVES: In this study, we investigated the effects of camphor on sperm quality in mice, and to protect sperm damage vitamin E as an antioxidant was used. METHODS: This study was conducted on 30 adult male mice (balb/c) with weight range 20-25 gr in 5 groups. First group was control (CO) and treated with normal saline, groups 2 and 3 were sham groups treated respectively with Olive oil (OL) and the combination of olive oil and vitamin E (OL+E), and finally, two experimental groups were treated using camphor (CA) and camphor with vitamin E (CA+E). Camphor at doses of 30 mg/kg/day and vitamin E at doses of 100 mg/kg/day were prepared. All materials were administered orally (gavage). After 35 days semen were collected from tail of epididymis, and then total count, motility, viability, nuclear maturity, and DNA damage were examined. RESULTS: Results showed significant reduction in sperm total count, percentage of viability, increase in the number of immature sperms and no significant difference in rate of motile sperms and sperms with damaged DNA in groups that received Camphor was observed. Vitamin E as a strong antioxidant administered lightly was able to reduce the effects of Camphor on viable and mature sperms (p<0.05). CONCLUSIONS: It can be concluded that Camphor could affect on mice sperm quality and vitamin E as an antioxidant, was able to slightly reduce Camphor effects in sperm quality.
Show more [+] Less [-]Hematological and biochemical reference values for C57BL/6, Swiss Webster and BALB/c mice
2016
Ed Wilson Santos | Dalila Cunha de Oliveira | Araceli Hastreiter | Graziela Batista da Silva | Jackeline Soares de Oliveira Beltran | Maristela Tsujita | Amanda Rabelo Crisma | Silvânia Meiry Peris Neves | Ricardo Ambrósio Fock | Primavera Borelli
The use of animals in scientific research has contributed significantly to the development of science, promoting various advances in understanding the metabolic machinery and the discovery of treatments and preventive measures applied to human and veterinary medicine. The development and use of alternative methods is encouraged; however, in some situations, the use of animals in accordance with ethical policies is still required. Established hematological and clinical chemistry reference values in laboratory animals are essential to evaluate functional changes; however, there are few data in the literature on these values, being fundamentally a comparative basis. The aim of this investigation was the establishment of hematological and clinical chemistry reference values in common strains/stocks of mice used in animal experimentation. Blood profile (hemogram, reticulocytes and myelogram) and clinical chemistry serum determination of total protein, albumin, glucose, cholesterol, triglycerides, calcium and phosphorus were evaluated using C57BL/6, BALB/c and Swiss Webster mice, male, 2-3 months old. The results standardize reference intervals in animals reared in Laboratory Animal Facility, reflecting the expected condition in rodents subjected to scientific research.
Show more [+] Less [-]Chinese herbal Jin-Ying-Tang attenuates the inflammatory response by inhibiting the activation of TLR4/MyD88/TRAF-6/NIK pathway at the mRNA level in LPS-stimulated mouse mammary epithelial cells
2016
Yi, Qiong | Li, Xin | Li, Yuan-Fang | Yang, Hang | Zhang, Xiao-Yi | Ma, Zhe | Wang, Lu
Introduction: The effects of Jin-Ying-Tang (JYT) on Toll-like Receptor 4 (TLR4) signalling transduction of lipopolysaccharide (LPS)-stimulated mouse mammary epithelial cells (MECs) in vitro were examined. Material and Methods: The cytotoxicity of JYT (0.06-62.50 mg/mL) on mouse MECs was determined by MTT assay. The MECs were co-cultured with LPS in the presence or absence of JYT (39.10 μg/mL, 391 μg/mL, 3910 μg/mL). The concentrations of interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) in the culture supernatants were detected by ELISA. The mRNA expression of TLR4 and downstream TLR4 signalling molecules such as myeloid differentiation factor 88 (MyD88), tumour necrosis factor receptor associated factor 6 (TRAF-6), inhibitor κB (IκB), and nuclear factor κB inducing kinase (NIK) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Results: The results showed that the IC₅₀ of JYT on MECs was 12.25 mg/mL and JYT could significantly decrease the concentrations of IL-6 and TNF-α in LPS-stimulated MECs (P < 0.05). The mRNA expression of TLR4, MyD88, TRAF-6, IκB, and NIK was also significantly decreased when the LPS-stimulated MECs were cocultured at appropriate concentrations of JYT (P < 0.05, P < 0.01). Conclusion: These observations indicate a potential mechanism through which JYT attenuates the systemic inflammatory response to LPS-stimulated mouse mammary epithelial cells by inhibiting the activation of TLR4/MyD88/ TRAF-6/NIK pathway at the mRNA level.
Show more [+] Less [-]Mouse duodenum as a model of inflammation induced by enterotoxigenic Escherichia coli K88
2016
Introduction: The aim of the experiment was to establish the enterotoxigenic Escherichia coli K88 (ETEC K88)-induced BALB/c mouse duodenum inflammation model. Material and Methods: Mice were administered different concentrations of E. coli K88 (1.0 × 10⁷-10⁹ CFU/mL) for 3 d by means of an esophageal catheter. Results: The results showed that the treated group expressed several significant clinical symptoms, such as reduced dietary demands and weight loss, an increased presence of IL-1α, TNF-α, and MPO in the peripheral blood, and some pathological changes in the duodenum. On the 6ᵗʰ-8ᵗʰ days, the body weight of the mice was the lowest. On the 8ᵗʰ day, there were significant differences in IL-1α, TNF-α, and MPO levels compared to the control group (P < 0.05), the gap between the duodenum mucous layer and the muscular layer had widened, the number of goblet cells was increased, and the inflammatory infiltrate and inflammation changes in the lamina propria and the mucous layer were the most obvious. Conclusion: The duodenum inflammation was the most severe on day 8; thus, the model was successfully established. In addition, varying concentrations of ETEC K88 did not significantly influence the duodenum inflammation (P > 0.05).
Show more [+] Less [-]Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model
2016
Kim, Chan Song | Hur, Jin | Eo, Seong Kug | Park, Sang-Youel | Lee, John Hwa
Salmonella Typhimurium ghost cells expressing K88ab, K88ac, K99, and FasA fimbriae of enterotoxigenic Escherichia coli (ETEC) in their envelopes were constructed. The genes encoding the fimbriae were individually cloned into an expression plasmid, pMMP81, carrying the asd gene, which was subsequently electroporated into the Δasd S. Typhimurium mutant. Plasmid pJHLP99, carrying the phiX174 lysis gene E, was also subsequently electroporated into the Salmonella mutant. The presence of the individual fimbriae on the ghost cells was examined by Western blot analysis. Forty BALB/c mice were equally divided into 2 groups of 20 mice each. Group A mice were intramuscularly vaccinated with a mixture of the 4 ghost cells expressing the individual fimbriae. The group B mice were inoculated with sterile phosphate-buffered saline as a control. The antigen-specific serum IgG concentrations were significantly higher in group A than in group B from week 2 until week 6 after inoculation. In addition, the antigen-specific IgA concentrations in fecal samples were significantly higher in group A than in group B at week 2 after inoculation. A large difference between the groups in the number of antigen-specific IgA-secreting cells in the small intestine was observed by immunohistochemical study. Also, the splenic lymphocyte proliferative responses were significantly greater in group A than in the control mice. These results suggest that vaccination with our Salmonella ghost cells can induce both humoral and cell-mediated immune responses and that the increased number of antigen-specific IgA-secreting cells in the small intestine may be correlated with the elevated fecal IgA immune response.
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