母体肢体缺血预处理对宫内窘迫胎鼠复氧后海马神经元线粒体结构和功能的影响

发布时间：2018-04-01 11:08

本文选题：肢体缺血预处理　切入点：胎儿缺氧/复氧损伤　出处：《福建医科大学》2014年硕士论文

【摘要】：目的探索母体肢体缺血预处理（LIP）对宫内窘迫胎鼠复氧后海马神经元线粒体结构和功能的影响。方法孕20天的SD大鼠麻醉后开腹采用微动脉夹钳夹母鼠通向子宫和卵巢的动静脉15min制备胎鼠宫内窘迫模型；同时将孕鼠右下肢股动脉阻断5min/再灌注5min，共3次循环进行母体的LIP。将24只孕鼠随机分为4组，每组6只：空白对照组（S组），LIP对照组，胎儿宫内窘迫组（FD组），LIP+胎儿宫内窘迫组。孕21天时剖宫取活胎鼠断头取脑，每只孕鼠取6只活胎鼠。通过电镜下观察胎鼠海马CA1区线粒体超微结构的变化；流式细胞仪检测海马线粒体跨膜电位、海马组织活性氧（ROS）的变化；定磷法检测海马组织ATP含量变化；比色法测定Mn-SOD活性及MDA含量。结果(1)与S组比较，LIP组电镜下观察胎鼠海马CA1区线粒体结构基本正常，无明显变化。线粒体膜电位、活性氧含量、ATP含量、Mn-SOD活性及MDA含量无明显变化（均P0.05）。（2）与S组比较，FD组电镜下观察胎鼠海马CA1区线粒体呈高度肿胀、部分脊断裂、消失，甚至呈空泡状，线粒体数目减少。线粒体膜电位下降、活性氧含量增加、ATP含量下降、Mn-SOD活性降低、MDA含量增加（均P0.05）。（3）与S组相比，LIP+FD组胎鼠海马CA1区线粒体呈轻度肿胀，部分脊断裂，线粒体外膜完整，线粒体膜电位下降、活性氧含量增加、ATP含量下降、Mn-SOD活性降低、MDA含量增加（均P0.05）。（4）与FD组比较，LIP+FD组胎鼠海马CA1区线粒体的超微结构保持较好的完整性，肿胀度减轻，抑制了线粒体膜电位的降低，减少海马ATP含量下降，Mn-SOD活性显著增高，，MDA含量减少（均P0.05）。结论母体肢体缺血预处理可抑制宫内窘迫胎鼠复氧后海马神经元线粒体超微结构的改变，减轻线粒体膜电位的下降，减少海马组织ATP含量降幅，减少ROS生成，增强线粒体抗氧化能力，改善线粒体功能。
[Abstract]:Objective to investigate the effects of maternal limb ischemic preconditioning (LIP) on the mitochondrial structure and function of hippocampal neurons in fetal rats with intrauterine distress after reoxygenation. Methods Sprague-Dawley rats were anesthetized on the 20th day of gestation and intrauterine distress model was established by using arteriovenous 15min (15min) which was clamped by arteriovenous clamp to the uterus and ovary. At the same time, the femoral artery of the right lower extremity of pregnant rats was blocked by 5min/ reperfusion for 5 minutes, and the maternal lip was carried out 3 times. 24 pregnant rats were randomly divided into 4 groups, 6 rats in each group: the blank control group (S group) and the control group (group S). Fetal intrauterine distress group (FD group) and lip fetal distress group (FID group). At 21 days of gestation, the fetal rat head was cut off and the brain was taken from each pregnant rat. The ultrastructure of mitochondria in CA1 area of fetal rat hippocampus was observed by electron microscope. The changes of mitochondrial transmembrane potential and reactive oxygen species (Ros) in hippocampal tissue were detected by flow cytometry, ATP content in hippocampal tissue was detected by phosphorus determination method, Mn-SOD activity and MDA content in hippocampus were measured by colorimetric method. Results 1) compared with S group, the mitochondrial structure in the hippocampal CA1 area of fetal rats was basically normal, and the mitochondrial membrane potential was not changed under electron microscope in lip group. There was no significant change in the activity of Mn-SOD and the content of MDA (P0.05. 0. 0. 0. 0. 0. 0. 5%) compared with group S, the mitochondria in CA1 area of fetal rat hippocampus were highly swollen, some ridges were broken, disappeared, or even vacuolated under electron microscope in FD group. The number of mitochondria decreased, the mitochondrial membrane potential decreased, the content of reactive oxygen species increased, the content of ATP decreased, the activity of Mn-SOD decreased, the activity of Mn-SOD decreased and the content of MDA increased. Compared with group S, the mitochondria of CA1 area of fetal rats in lip FD group were slightly swollen and some ridges were broken. The mitochondrial outer membrane was intact, the mitochondrial membrane potential was decreased, the content of reactive oxygen species was increased, the activity of Mn-SOD was decreased, and the content of MDA was increased (P0.05, P < 0.05). Compared with the FD group, the ultrastructure of mitochondria in the CA1 area of fetal rats in the lip FD group maintained a better integrity than that in the FD group. The swelling degree was reduced, the decrease of mitochondrial membrane potential was inhibited, the content of ATP in hippocampus was decreased, the activity of Mn-SOD was increased significantly and the content of MDA was decreased (all P 0.05). Conclusion the maternal limb ischemic preconditioning can inhibit the changes of mitochondrial ultrastructure of hippocampal neurons after reoxygenation in fetal rats with intrauterine distress, reduce the decrease of mitochondrial membrane potential, decrease the content of ATP and decrease the production of ROS in hippocampus. Enhance the antioxidant capacity of mitochondria and improve the function of mitochondria.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R614

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