RESUMO
Amniotic fluid is a rich source of multipotent mesenchymal stem cells (MSCs). Amniotic fluid stem cells (AFSCs) have become a new source of stem cells; they have low immunogenicity and are easily harvested. For this reason, they may be useful in clinical tissue engineering. Moreover, AFSCs have anti-inflammatory properties and can repair tissues. This study evaluated the utility of AFSC injection to treat bilateral ovarian dystrophy in Holstein-Friesian cows. Bovine AFSCs (BAFSCs) were collected at slaughter from Holstein-Friesian cows during the third or fourth month of pregnancy and cultured in vitro. The BAFSCs began to show a fibroblast-like morphology. They were positive for ß-integrin, CD44, CD73, CD106 and Oct4 and negative for CD34 and CD45. After induction, the cells differentiated into mesodermal lineages. Bilateral ovarian dystrophy was confirmed by ultrasonography in 16 lactating cows. The subsequent experiment lasted 15 weeks. Serum was collected weekly to analyse progesterone concentrations, and weekly ultrasonography recorded ovarian changes. Each cow was equipped with an automatic heat detection system to facilitate oestrus observation and breeding records. The progesterone concentration of two cows in the treatment group (25%) significantly increased during weeks 10-15. On ultrasonography, the treatment group demonstrated mature follicles after BAFSCs injection, and foetuses were visualized approximately 40 days after artificial insemination (AI). Oestrus rates in the control and treatment groups were 0% (0/8) and 50% (4/8), respectively; pregnancy rates were 0% (0/8) and 25% (2/8), respectively. Calves were successfully delivered in both cases of pregnancy. These results show that BAFSCs can alleviate bovine ovarian dystrophy and restore fertility.
Assuntos
Líquido Amniótico/citologia , Doenças dos Bovinos/terapia , Transplante de Células-Tronco Mesenquimais , Doenças Ovarianas/veterinária , Animais , Bovinos , Diferenciação Celular , Células Cultivadas , Clima , Feminino , Fertilidade , Inseminação Artificial/veterinária , Células-Tronco Multipotentes/transplante , Doenças Ovarianas/terapia , Gravidez , Progesterona/sangueRESUMO
OBJECTIVE: To investigate the protection of Verapamil against advanced glycation end products (AGE) induced human lens epithelial cells (HLEC) apoptosis. METHODS: Experiment study. SRA01/04 (HLEC line) was cultivated and passaged to the third generation and then divided into four groups. A group was named as control group, and B group was named as AGE group (LEC was treated by 20 µmol/L AGE). C group was AGE+SB202190 group (LEC was treated 2 hours by SB2012190 and then treated by 15 µmol/L AGE). D group was AGE+ Verapamil group (LEC was treated 2 hours by 50 µmol/L Verapamil and then treated by AGE). MTT was used to evaluate the cell viability. Flow cytometry with Annexin V-FITC apoptosis detection was used to assess cell apoptosis.The expression of p-p38 and caspase3 was detected by Western blot between groups. One way Chi-square analysis was used for data analysis. LSD-t test was used as comparison between every two groups. RESULTS: After 24 hours, LEC viability (A570) was (0.28±0.08) in B group, which was significantly lower than A group (0.97±0.05) (LSD-t test, P=0.008). LEC viability in C and D group was (0.79±0.06) and (0.62±0.07) separately, which can partly higher than it was in B group (F=34.52, P=0.001). The apoptosis cells were (19.9±1.1)% in B group, which were significantly higher than they were in A group (2.5±0.6)% (P=0.003). The apoptosis cells in C and D group were (4.23±1.20) and (5.79±1.75) separately, which were significant lower than they were in B group (F=371.61, P<0.01). In additional, expressions of p-p38, Caspase3 proteins in the cells of group B were (223.35±20.15) and (256.77±19.88) separately, which were higher than it were in A group,which were (106.44±10.74) and (100.26±18.65) separately. However, they were (139.17±19.10) and (142.75±23.36) in group C and (154.79±21.87) and (139.79±25.73) in group D (F=248.01, F=76.68; P<0.01), which were lower than they were in A group. CONCLUSION: p38 pathway is involved in the apoptotic procedure of LEC induced by AGE. Verapamil can interdict the p38 signal pathway and protect LEC apoptosis. (Chin J Ophthalmol, 2016, 52: 444-448).
Assuntos
Apoptose/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Produtos Finais de Glicação Avançada , Cristalino/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases , Verapamil/farmacologia , Caspase 3/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Distribuição de Qui-Quadrado , Citometria de Fluxo , Humanos , Cristalino/citologia , Cristalino/metabolismo , Transdução de Sinais , Fatores de TempoRESUMO
1. Mitochondria isolated from rats treated with glucagon for 60 min or lives perfused in the presence of glucagon for 10 min exhibited lower rates of 45Ca2+ exchange than did control mitochondria when this was measured under steady-state conditions in the presence of Mg2+, ATP, Pi and 0.13 microM- or 0.16 microM-free Ca2+ at pH 7.4 and at 25 degrees C or 37 degrees C. Under these conditions no significant difference in the rates of Ruthenium Red-induced 45Ca2+ efflux was observed. These results contrast with earlier work in which mitochondria isolated from glucagon-treated livers were shown to exhibit faster rates of Ca2+ uptake [Yamazaki (1975) J. Biol. Chem. 250, 7924-7930] and slower rates of spontaneous Ca2+ efflux [Hughes & Barritt (1978) Biochem. J. 176, 295-304] when these parameters were measured under different incubation conditions, including supra-physiological concentrations of free Ca2+ and the absence of added Mg2+ and ATP. 2. Perfusion of livers with glucagon before the addition of adrenaline or the Ca2+-selective ionophore A23187, to release Ca2+ from intracellular stores, decreased the amount of Ca2+ released by these agents. 3. Incubation of isolated hepatocytes in the presence of glucagon at 1.3 mM extracellular Ca2+ induced a small decrease in the plateau of the 45Ca2+-exchange curve obtained under steady-state conditions. 4. It is concluded that the actions of glucagon on liver mitochondrial Ca2+ transporters lead to a decrease, rather than an increase, in mitochondrial Ca2+ stores in the intact cell.