钒诱导蛋鸡输卵管膨大部上皮细胞氧化应激模型的建立

doi:10.11843/j.issn.0366-6964.2017.02.018

Abstract

Abstract:

This study was conducted to establish a V⁵⁺-induced oxidative stress model by using primary oviduct magnum epithelial cells (OMECs) of laying hens. OMECs were isolated from Lohman layers (2-3 weeks before onset of laying), cultured and characterized subsequently. Cultured OMECs were then treated with 0, 25, 50, 100, 250, and 1 000 μmol•L^-1 vanadium for 12 h to measure cell viability, apoptosis ratio as well as cellular ROS, SOD, MDA, CAT and LDH released. The results showed that: 1) cell island formed after 24 h and showed classical flagstone morphology. Cultured cells were positive to anti-CK18, anti-OVA, anti-ESR1 and anti-PGR antibody, while were negative to anti-vimentin antibody. 2) Cell viability rate were （69.90±2.78）% and （63.60±1.57）% after 50 and 100 μmol•L^-1 vanadium treated for 12 h, respectively, which were lower (P<0.05) than that of 25 μmol•L^-1 and higher (P<0.05) than that of 250 and 1 000 μmol•L^-1 vanadium. 3) The apoptosis rate increased(P<0.05) as vanadium levels increased, with the 1 000 μmol•L^-1 vanadium treatment had the highest apoptosis rate. 4) The cellular ROS, FITC mean values in 100 and 250 μmol•L^-1 vanadium groups were significantly higher than in 0 μmol•L^-1 vanadium treatment (P<0.05), meanwhile FITC mean value in 250 μmol•L^-1 was significantly higher than in 100 μmol•L^-1 vanadium treatment (P<0.05). 5) Compared with 0 μmol•L^-1 vanadium, cells treated with 50 μmol•L^-1 vanadium had lower (P<0.05) SOD activity, while 100, 250 and 1 000 μmol•L^-1 vanadium groups significantly increased MDA contents(P<0.05). The activity of CAT in 50, 100, 250 and 1 000 μmol•L^-1 vanadium treatments were significantly lower than in 0 μmol•L^-1 vanadium treatment (P<0.05). The amount of released LDH increased as vanadium levels increased, with the 1 000 μmol•L^-1 vanadium treatment had the highest value (P<0.05), and 100 μmol•L^-1 vanadium group had higher value than in 0 μmol•L^-1 (P<0.05), up to 141.61±2.81 U•gprot^-1. These results indicated that the primary OMECs were cultured successfully and supplementation of 100 μmol•L^-1 vanadium in primary OMECs can be used to establish an oxidative stress model in OMECs.

CLC Number:

S831
S85

HUANG Xuan-yang，WANG Jian-ping，DING Xue-mei，BAI Shi-ping，ZENG Qiu-feng，ZHANG Ke-ying. Establishment of an Oxidative Stress Model Induced by Vanadium in Oviduct Magnum Epithelial Cell of Laying Hens[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 340-350.

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Establishment of an Oxidative Stress Model Induced by Vanadium in Oviduct Magnum Epithelial Cell of Laying Hens

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