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Abstract

The aim of the present study was to determine the effects of Luteolin (LUT) on semen quality, oxidative stress, apoptosis, acrosomal integrity, mitochondrial membrane potential and dead sperm ratio in rabbits. Ejaculates from six New Zealand rabbits were collected, evaluated and pooled. The pooling was divided into five groups as control (no additive) LUT 25 μM, LUT 50 μM, LUT 100 μM and LUT 200 μM and LUT added. It was then filled into a falcon tube with Tris-based extender at a final concentration of approximately 35 x 106 spermatozoa. Diluated rabbit semen samples were drawn into frozen and thawed. Frozen semen straws were thawed at 37°C in 30 seconds. According to our findings, no statistical difference was found between all doses of luteolin and the control group in the CASA (computer assisted sperm analysis) analysis performed at 4°C. However, total motility, progressive motility and rapid sperm percentage were found to be higher in the frozen and thawed rabbit semen at a dose of LUT 50 μM compared to the other groups (p<0.05). While amplitude of lateral head displacement (ALH) and beat cross-frequency (BCF) values were found at the lowest dose of LUT 200 μM, a statistically significant difference was observed between the other groups. When the flow cytometry results were examined, no statistical difference was found between the rate of dead sperm, acrosomal integrity, mitochondrial membrane potential and apoptosis rate. Morever, the H2O2 percentage was found to be lower in all experimental groups compared to the control group (p<0.001). In conclusion, the addition of LUT in long-term storage of rabbit semen provided a protective effect for spermatozoa with its antioxidative properties against damage caused by cryopreservation.
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Authors and Affiliations

S.A. Akarsu
1
T.C. Acısu
2
İ.H. Güngör
2
A. Çakır Cihangiroğlu
2
R.H. Koca
3
G. Türk
2
M. Sönmez
2
S. Gür
2
F. Fırat
2
H.E. Esmer Duruel
4

  1. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Atatürk University, 25240 Yakutiye /Erzurum, Turkey
  2. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Fırat University, 23200, Elazığ, Turkey
  3. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Bingöl University, Selahaddin-i Eyyübi Neighbourhood, University Street No: 1, 12000, Bingöl, Turkey
  4. Elbistan Vocational School, Kahramanmaraş İstiklal University, Şehit Astsubay Ömer Halisdemir Street No:5, Doğan Neighbourhooh,46340, Elbistan, Kahramanmaraş, Turkey
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Abstract

The present study was undertaken to highlight the influence of simvastatin administration on hepatocyte morphology, proliferation, and apoptosis. The study included 48 gilts aged 3 months (weighing ca. 30 kg) divided into groups I (control; n=24) and II, receiving 40 mg/animal simvas- tatin orally (simavastatin; n=24) for 29 days. The animals were euthanized on days subsequent to the experiment. The livers were sampled, fixed, and processed routinely for histopathology, histochemistry, and immunohistochemistry (for proliferating cell nuclear antigen, Bcl-2, and caspase-3). Apoptosis was visualized by terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL). Simvastatin administration caused acute hepatocyte swelling, glycogen de- pletion, hyperaemia, multifocal hepatocyte proliferation with occasional pseudoacinar formation, connective tissue hyperplasia, eosinophil infiltration, and interface hepatitis. The proliferating cell nuclear antigen index, mean diameter of argyrophilic nucleolar organizer regions, and Bcl-2 immunoexpression were lower compared to control, and mean caspase-3 immunoexpression was higher in group II compared to control. On day 25 and 29 single hepatocytes in the simvasta- tin-treated group were TUNEL-positive. Simvastatin caused morphological alteration which became more intense over time. The results from the present study suggest that simvastatin treat- ment may cause glycogen, lipid metabolism and cell membrane permeability distortion, fibrosis, interface hepatitis, reduction in hepatocyte proliferation and transcriptional activity, and enhanced vulnerability to apoptosis. Summing up the results, it can be concluded that simvastatin caused liver damage with similar morphological changes seen in autoimmune-like liver injury, which may indicate that simvastatin may induce autoimmune-like drug induced liver injury.

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Authors and Affiliations

M. Mikiewicz
I. Otrocka-Domagała
K. Paździor-Czapula
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Abstract

The aim of our study was to determine the influence of L-carnitine (L-CAR) on the cellular parameters of hen erythrocytes during a 48 hour exposure to L-CAR at concentrations of 25, 50 and 100 mg/mL in nutrient-deficient medium. Cell morphology, haemolysis, caspase 3/7 activity and glucose uptake (GU) were determined. The results showed a lower percentage of apoptotic cells and decreased haemolysis of erythrocytes treated for 48 hours at all the concentrations of L-CAR. The amino acid at 50 mg/mL inhibited the activity of proapoptotic caspase 3/7; however, it increased GU. In contrast, caspase 3/7 level was increased but GU was decreased in erythrocytes treated with 100 mg/mL of L-CAR when compared to the control. It may be hypothesized that reduction of apoptotic changes in hen erythrocytes may result from increased GU.

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Authors and Affiliations

K. Kasperek
A. Bownik
S. Knaga
A. Szabelak
B. Ślaska
M. Kwiecień
G. Jeżewska-Witkowska
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Abstract

Emerging researches in humans, pigs and mice, highlighted that estrogen plays a pivotal role in self-renewal and differentiation of bone marrow mesenchymal stem cells (BMSCs). The present study aimed at evaluating effects of 17 beta-estradiol (E2) on proliferation and apoptosis of canine-derived bone marrow mesenchymal stem cells (cBMSCs) in vitro. The results showed that E2 supplementation at the concentration of 10-11 M promoted the proliferation of cBMSCs by CCK-8 assay and RT-qPCR analysis for the proliferation-related genes, with proliferating cell nuclear antigen (PCNA), cyclin-D1 (CCND1) being up-regulated and cyclin-dependent kinase inhibitor 1B (CDKN1B) being down-regulated. Contrarily, analysis of fluorescence-activated cell sorting (FACS) and RT-qPCR demonstrated that E2 supplementation above 10-11 M had inhibitory effects on the proliferation of cBMSCs and induced apoptosis. Intriguingly, cBMSCs still possessed the capability to differentiate into osteoblasts and adipocytes with 10-11 M E2 addition. Taken together, this study determined the optimal culture condition of cBMSCs in vitro, and has important implications for further understanding the regulatory effect of E2 on the self-renewal of cBMSCs, which are helpful for the clinical application of BMSCs.

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Authors and Affiliations

Z.-H. Zhou
C.-W. Gu
J. Li
X.-Y. Huang
J.-Q. Deng
L.-H. Shen
S.-Z. Cao
J.-L. Deng
Z.-C. Zuo
Y. Wang
ORCID: ORCID
X.-P. Ma
Z.-H. Ren
S.-M. Yu

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