过表达硫氧还蛋白还原酶1对卵巢去势大鼠海马神经元的保护效应  

Protective effect of overexpression of thioredoxin reductase 1 on hippocampal cells of ovariectomized rats

作　　者：刘自华[1] 李晓娟[1] 赵贵玲[2] 王芳 Liu Zihua;Li Xiaojuan;Zhao Guiling;Wang Fang(Department of Transfusion Blood,Lanzhou University Second Hospital,Lanzhou 730030,China;The Department of Neurology,Lanzhou University Second Hospital,Lanzhou 730030,China;Gynecology with Integrated Traditional Chinese and Western Medicine,Gansu Provincial Maternity and Child-care Hospital,Lanzhou 730050,China)

机构地区：[1]兰州大学第二医院输血科,兰州730030 [2]兰州大学第二医院神经内科,兰州730030 [3]甘肃省妇幼保健院中西医结合妇科,兰州730050

出　　处：《中华行为医学与脑科学杂志》2022年第10期926-932,共7页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：甘肃省青年科学基金(17JR5JA227)。

摘　　要：目的探讨过表达硫氧还蛋白还原酶1(thioredoxin reductase 1,TR1)对卵巢去势大鼠海马神经元的影响。方法根据随机数字表法将54只雌性SD大鼠分为正常组、卵巢去势组和卵巢去势过表达TR1组(卵巢去势-TR1组),每组18只。用慢病毒构建过表达TR1载体,脑立体定位仪海马定位进行注射慢病毒重组体,通过qRT-PCR和Western blot检测大鼠海马组织TR1、Bcl-2,p53和p21的mRNA和蛋白水平;用Western blot检测海马Caspase-3的表达;用WST-1细胞增殖及细胞毒性检测试剂法检测海马超氧化物歧化酶(super oxide dimutase,SOD)活性、用微板法测定谷胱甘肽(glutathione,GSH)含量、用硫代巴比妥酸法测定丙二醛(malondialdehyde,MDA)含量;通过旷场实验和水迷宫实验检测大鼠行为学变化。用GraphPad Prism 9.0进行统计学处理,三组间比较采用单因素方差分析,进一步两两比较采用LSD检验;两组间比较采用独立样本t检验。结果(1)卵巢去势组大鼠海马组织TR1的mRNA及其蛋白水平均低于正常组大鼠(t=3.125,4.402,均P<0.05),卵巢去势-TR1组大鼠海马组织TR1的mRNA及其蛋白水平均高于卵巢去势组(t=4.945,4.845,均P<0.05)。(2)三组大鼠在水迷宫实验中的逃避潜伏期和旷场实验中的运动距离和中央区停留时间均差异有统计学意义(F=44.73,33.67,6.51,均P<0.05)。在旷场实验中,卵巢去势大鼠运动距离多于正常组[(4700±141)mm,(3967±163)mm],中央区停留的时间长于正常组[(87.33±3.93)s,(80.83±2.48)s],卵巢去势-TR1组的运动距离[(4267±150)mm]和中央区停留时间[(82.17±3.43)s]均低于卵巢去势组(均P<0.05)。在水迷宫实验中,卵巢去势组大鼠的逃避潜伏期高于对照组[(28.67±2.50)s,(19.50±2.59)s,P<0.05],而卵巢去势-TR1组逃避潜伏期低于卵巢去势组[(25.00±1.67)s,P<0.05]。(3)三组大鼠海马组织氧化应激损伤指标MDA、SOD和GSH的水平均差异有统计学意义(F=87.41,91.38,28.69,均P<0.01)。卵巢去势组大鼠海�Objective To investigate the effects of thioredoxin reductase 1(TR1)overexpression on hippocampus in ovariectomized SD rats.Methods Totally 54 female SD rats were divided into normal group,ovariectomized group and ovariectomized over-expressioned TR1 group(ovariectomy-TR1 group)according to the random number table method with 18 in each group.The overexpressed TR1 vector was constructed by lentivirus,and the recombinant lentivirus was injected into the hippocampus by a brain stereotactic instrument.The mRNA levels and protein levels of TR1,Bcl-2,p53 and p21 in the hippocampus of SD rats were detected by qRT-PCR and Western blot.The expression of Caspase-3 in the hippocampus of SD rats was detected by Western blot.The activity of SOD was measured by the WST-1 cell proliferation and cytotoxicity method,the content of GSH was measured by the microplate method,and the content of MDA in the hippocampus of SD rats was measured by the TBA method.The behavioral changes of SD rats were detected by the open field test and water maze test.GraphPad Prism 9.0 was used for statistical analysis.One-way analysis of variance was used for comparisons among the three groups,and the LSD test was used for further pairwise comparisons,the t-test was used to compare the mean number of samples between the two groups.Results(1)The mRNA and protein levels of TR1 in hippocampus of ovariectomized rats were lower than those of normal rats(t=3.125,4.402,both P<0.05).The mRNA and protein levels of TR1 in hippocampus of ovariectomized-TR1 rats were higher than those of ovariectomized rats(t=4.945,4.845,both P<0.05).(2)There were significant differences among the three groups in the escape latency in water maze test,the movement distance and the stay time in central area in the open field test(F=44.73,33.67,6.51,all P<0.05),the movement distance in the open field test of ovariectomized rats was more than that of the normal group((4700±141)mm,(3967±163)mm,P<0.05),the stay time in the central area was longer than that of the normal group((87.33�

关 键 词：卵巢去势 硫氧还蛋白还原酶1 学习和记忆 凋亡 大鼠 

分 类 号：R338[医药卫生—人体生理学]