梭曼急性暴露对大鼠肝脏线粒体的损伤作用研究  

Damage of acute soman exposure to liver mitochondria in rats

作　　者：靳倩 崔雅岚 时梦 张毅 朱思庆 石晶晶 陈学军 李丽琴 JIN Qian;CUI Ya-lan;SHI Meng;ZHANG Yi;ZHU Si-qing;SHI Jing-jing;CHEN Xue-jun;LI Li-qin(State key Laboratory of NBC Protection for Civilians,Beijing 102205,China)

机构地区：[1]国民核生化灾害防护国家重点实验室,北京102205

出　　处：《军事医学》2023年第4期268-272,292,共6页Military Medical Sciences

摘　　要：目的研究梭曼急性暴露诱导大鼠肝脏线粒体损伤作用及其肝毒性机制,为抗毒药物的开发提供理论依据。方法20只SPF级雄性SD大鼠,随机分为对照组(6只)和梭曼染毒组(14只)。梭曼染毒组大鼠皮下注射梭曼98μg/kg(1×LD50),对照组大鼠皮下注射等体积生理盐水。梭曼染毒30 min后,取其中的6只存活大鼠和对照组6只大鼠分别断头处死,解剖肝脏进行分析;利用透射电镜检测肝脏组织线粒体形态损伤情况;采用比色法检测各组肝组织乙酰胆碱酯酶(AChE)活性和乙酰胆碱(ACh)水平;应用生化试剂盒检测肝组织氧化应激标志物,即丙二醛(MDA)、过氧化脂质(LPO)含量及谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)的水平;采用实时高分辨呼吸测定系统检测大鼠肝脏线粒体呼吸功能。结果大鼠梭曼染毒30 min后,肝组织AChE活性显著降低(P<0.01),且ACh水平显著升高(P<0.01);大鼠肝脏线粒体发生肿胀,染色质减少,嵴消失,且在细胞核周围出现糖原聚集;大鼠肝脏LPO、MDA和GSSG水平显著升高(P<0.05),且GSH水平降低(P<0.01)。线粒体呼吸功能检测结果显示,梭曼染毒组大鼠肝脏的基础呼吸水平(P<0.05)、ATP产量(P<0.01)和非线粒体耗氧量(P<0.05)显著降低。同时,梭曼还降低了线粒体复合物Ⅰ、复合物Ⅱ、复合物Ⅰ+Ⅱ活性(P<0.05)。结论大鼠急性梭曼暴露可导致肝脏线粒体在生物氧化和能量转换过程中产生大量活性氧,线粒体发生氧化应激,进而导致线粒体能量代谢失调,表现为破坏线粒体的氧化磷酸化,干扰线粒体呼吸链,抑制ATP合成,最终诱导肝细胞损伤。Objective To investigate the liver injury induced by acute exposure to soman in rats and the hepatotoxic mechanism in order to provide data for the development of antitoxic drugs.Methods Twenty SPF grade male SD rats were randomly divided into the control group(6 rats)and soman-exposed group(14 rats).Rats in the soman-exposed group were subcutaneously injected with 98μg/kg(1×LD50)while those in the control group were subcutaneously injected with an equal volume of normal saline.After 30 minutes′exposure to soman,six surviving soman-exposed rats and six control rats were decapitated and their livers were dissected for analysis.The morphological damage to liver mitochondria was detected under a transmission electron microscope(TEM).The activity of acetylcholinesterase(AChE)and the level of acetylcholine(ACh)in liver tissue of each group were detected by colorimetry.The contents of malondialdehyde(MDA),lipid peroxide(LPO)and the levels of glutathione(GSH)and oxidized glutathione(GSSG)in liver tissue were detected by biochemical assay kits.A real-time high-resolution respirometry system(Oxygraph-2k)was used to detect the oxygen consumption associated with mitochondrial respiratory function in rat livers.Results After 30 minutes'exposure to soman,AChE activity decreased significantly,while the level of ACh increased significantly in liver tissue.Mitochondria of rat livers swelled,chromatin decreased,the ridges disappeared,and glycogen was accumulated around the nucleus.The levels of LPO,MDA and GSSG in rat livers increased significantly,but the level of GSH decreased.The results of mitochondrial respiratory function showed that basal respiration,ATP production and non-mitochondrial oxygen consumption of livers of soman-exposed rats decreased significantly.Meanwhile,soman reduced the activities of mitochondrial complexesⅠ,Ⅱ,Ⅰ+Ⅱ.Conclusion Acute soman exposure in rats can cause liver mitochondria to produce a large amount of reactive oxygen species(ROS)in the process of biological oxidation and energy conver

关 键 词：梭曼 肝损伤 线粒体 氧化应激 呼吸链 

分 类 号：R827.12[医药卫生—临床医学]