Guo Yuanbo, Wang Yan, Zhang Dengwen, Cui Can, Li Tao, Wang Sheng
Department of Anesthesiology of Medical Sciences, Guangzhou 510080, China.
Department of Science and Education, Guangdong Provincial People's Hospital/Guangdong Academy of Medical Sciences, Guangzhou 510080, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2019 Jul 30;39(7):850-854. doi: 10.12122/j.issn.1673-4254.2019.07.16.
To investigate the effect of ulinastatin pretreatment on isoflurane-induced mitochondria-dependent neuronal apoptosis in the hippocampus of rats.
Thirty-six male SD rats were randomly assigned into control group, isoflurane group and ulinastatin group. In the latter two groups, the rats were subjected to acute exposure to 0.75% isoflurane for 6 h and pretreated with 50 000 U/kg of ulinastatin before isoflurane exposure, respectively. After the treatments, apoptosis of the hippocampal neurons was detected using TUNEL assay, and the mitochondrial membrane potential (△ ψm) was measured using JC-1 mitochondrial membrane potential kit; cytochrome C release and caspase-3 activity were examined with Western blotting, and intracellular reactive oxygen species (ROS) was detected using the fluorescent probe H2DCFDA.
Compared with those in the control group, the rats with acute exposure to isoflurane showed markedly increased TUNEL-positive cells in the hippocampus ( < 0.05), which were obviously reduced by ulinastatin pretreatment ( < 0.05). The △ψm of the hippocampal neurons was significantly reduced after isoflurane exposure ( < 0.05), and was partly recovered by ulinastatin pretreatment ( < 0.05). Acute exposure to isoflurane resulted in obviously increased cellular ROS, cytochrome C release and caspase-3 activity in the hippocampal neurons ( < 0.05), and these changes were significantly inhibited by ulinastatin pretreatment ( < 0.05).
Ulinastatin pretreatment provides neuroprotection against isoflurane-induced apoptosis of the hippocampal neurons in rats possibly by inhibiting mitochondria-dependent apoptosis pathway.
探讨乌司他丁预处理对异氟烷诱导的大鼠海马线粒体依赖性神经元凋亡的影响。
将36只雄性SD大鼠随机分为对照组、异氟烷组和乌司他丁组。后两组大鼠分别急性暴露于0.75%异氟烷6小时，并在异氟烷暴露前用50000 U/kg乌司他丁进行预处理。处理后，采用TUNEL法检测海马神经元凋亡，用JC-1线粒体膜电位试剂盒检测线粒体膜电位（△ψm）；用蛋白质免疫印迹法检测细胞色素C释放和半胱天冬酶-3活性，用荧光探针H2DCFDA检测细胞内活性氧（ROS）。
与对照组相比，急性暴露于异氟烷的大鼠海马中TUNEL阳性细胞明显增多（<0.05），乌司他丁预处理可明显减少（<0.05）。异氟烷暴露后海马神经元的△ψm显著降低（<0.05），乌司他丁预处理可部分恢复（<0.05）。急性暴露于异氟烷导致海马神经元细胞内ROS、细胞色素C释放和半胱天冬酶-3活性明显增加（<0.05），乌司他丁预处理可显著抑制这些变化（<0.05）。
乌司他丁预处理可能通过抑制线粒体依赖性凋亡途径对异氟烷诱导的大鼠海马神经元凋亡提供神经保护作用。
