[目的]采用脂多糖(LPS)作为刺激源,建立奶牛子宫内膜上皮细胞(BEEC)氧化损伤模型.[方法]体外培养BEEC并进行细胞鉴定,用不同浓度的LPS刺激细胞,在不同时间点用CCK-8法测定细胞存活率,以确定BEEC氧化损伤模型条件.倒置显微镜下观察细胞形态学变化,流式细胞术检测细胞凋亡率,DCFH-DA检测细胞内活性氧(ROS)含量,比色法检测细胞中的丙二醛(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT).[结果]与空白对照组相比,当LPS浓度为80μg/mL,作用时间为12 h时,造成细胞损伤,其细胞凋亡率、ROS产生量和MDA含量明显高于对照组,SOD和CAT活性显著低于对照组.[结论]建立奶牛子宫内膜上皮细胞氧化损伤模型的条件为LPS作用浓度80 μg/mL,作用时间12 h.%[Objective]To establish the oxidative injury model of bovine endometrial epithelial cell (BEEC),lipopolysaccharide (LPS) was used as stress source.[Methods]BEECs were stimulated with different concentration of LPS in vitro,and the cells viability were measured by the method of CCK-8 at different time to determine the conditions of oxidative injury model.Cellular morphology were observed by inverted microscope.The apoptosis rate of cells was detected by flow cytometry.The intracellular reactive oxygen species (ROS) content was detected by DCFH-DA.The malondialdehyde (MDA),superoxide dismutase (SOD) and catalase (CAT) were detected by the colorimetric method.[Results]Compared with control group,cells were injured at the concentration of 80 μg/mL LPS for 12 h.The apoptosis rate,ROS and MDA content were significantly higher,SOD and CAT activity were significantly lower.[Conclusion]The oxidative injury model in BEEC was established using 80 μg/mL LPS for 12 h.
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