海马注射脑源性神经营养因子对大鼠脑卒中后抑郁的作用及机制  

Effect and mechanism of brain-derived neurotrophic factor injected into hippocampus on post-stroke depression in rats

作　　者：张珍珍 位慧芳 张勇 张辉 ZHANG Zhenzhen;WEI Huifang;ZHANG Yong;ZHANG Hui(Department of Neurology,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou,Henan 450000,China;Department of Neurosurgery,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou,Henan 450000,China)

机构地区：[1]郑州大学附属郑州中心医院神经内科,河南郑州450000 [2]郑州大学附属郑州中心医院神经外科,河南郑州450000

出　　处：《安徽医药》2023年第8期1507-1511,I0002,共6页Anhui Medical and Pharmaceutical Journal

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20191045)。

摘　　要：目的观察海马内注射脑源性神经营养因子(BDNF)对大鼠脑卒中后抑郁的改善作用,并探究其对环磷酸腺苷/环磷酸腺苷反应元件结合蛋白/脑源性神经营养因子(cAMP/CREB/BDNF)信号通路的影响。方法2022年1―3月选择SPF级SD雄性大鼠95只,采用随机数字表法选择80只制备脑卒中后抑郁(PSD)模型,余15只为假手术组。将PSD造模成功的64只大鼠采用随机数字表法分为PSD组、无序干扰组、BDNF干预组、激活剂组和抑制剂组。造模成功2 h后,给予无序干扰组和BDNF组大鼠分别注射含无序干扰序列的慢病毒载体和含BDNF的慢病毒载体,激活剂组和抑制剂组分别给予皮下注射forskolin和SQ22536,假手术组和PSD组注射等量含5%二甲基亚砜的生理盐水。采用糖水实验和敞箱实验评估大鼠抑郁情况,采用实时荧光定量逆转录PCR(RT-qPCR)检测各组海马组织BDNF相对表达水平,苏木精-伊红(HE)染色观察海马组织病理学变化,测定各组大鼠海马组织去甲肾上腺素(NE)和5-羟色胺(5-HT)含量,蛋白质印迹法(Western blotting)检测海马区cAMP、pCREB、BDNF蛋白表达水平。结果与假手术组比较,PSD组、无序干扰组、BDNF组糖水消耗量[(42.69±2.18)%、(42.84±2.29)%、(71.66±3.04)%比(90.34±3.28)%]、方格穿行次数[(13.72±0.97)次、(13.98±0.75)次、(23.17±1.47)次比(39.66±1.45)次]和竖起修饰次数[(9.14±0.45)次、(9.52±0.53)次、(12.96±0.75)次比(18.65±1.12)次]减少,BDNF相对表达量(0.62±0.13、0.69±0.09、1.68±0.16比2.62±0.32)降低,且BDNF组高于PSD组和无序干扰组(P<0.05);HE染色结果显示,PSD组和无序干扰组海马神经元细胞形态相似,数目减少,排列紊乱;BDNF组海马神经元细胞数目增加,排列基本紧密,形态基本规则;与假手术组比较,PSD组、无序干扰组、BDNF组NE、5-HT水平降低,且BDNF组高于PSD组和无序干扰组(P<0.05)。与假手术组比较,PSD组、无序干扰组、BDNF组、抑制剂组和激活剂�Objective To observe the effect of intra-hippocampal injection of BDNF on post-stroke depression in rats,and explore its effect on cAMP/CREB/BDNF signaling pathway.Methods From January to March 2022,95 male SD rats with SPF grade were selected and 80 rats were randomly selected to prepare PSD model,and the remaining 15 rats were sham operated.The 64 rats that successfully established the post-stroke depression(PSD)rat model were randomly divided into PSD group,disordered interference group,BDNF group,activator group,and inhibitor group.Two hours after successful modeling,disordered interference group and BDNF group were injected with disordered interference sequence-containing lentiviral vector and BDNF-containing lentiviral vector,activator group and inhibitor group were injected subcutaneously with forskolin and SQ22536,sham operation group and PSD group were injected with equal amounts of 5%dimethyl sulfoxide in saline.Sugar water test and open box test were used to evaluate the depression of rats,and the relative expression levels of BDNF in hippocampus of each group were detected by RT-qPCR.Hippocampal histopathological changes were observed by HE staining.NE And 5-HT content,Western blot was used to detect the expression levels of cAMP,pCREB,and BDNF proteins in the hippocampus.Results Compared with sham operation group,the consumption of sugar water[(42.69±2.18)%,(42.84±2.29)%,(71.66±3.04)%vs.(90.34±3.28)%],the number of square passes[(13.72±0.97),(13.98±0.75),(23.17±1.47)vs.(39.66±1.45)]and erect modification[(9.14±0.45),(9.52±0.53),(12.96±0.75)vs.(18.65±1.12)],the relative expression of BDNF is reduced in the PSD group(0.62±0.13,0.69±0.09,1.68±0.16 vs.2.62±0.32),disorder interference group,BDNF group,and BDNF group>PSD group and disorder interference group(P<0.05).The results of HE staining showed that the hippocampal neuron cells of the PSD group and the disordered interference group had similar morphology,decreased in number and disordered arrangement;the number of hippocampal neurons i

关 键 词：脑源性神经营养因子 卒中 抑郁症 脑梗死 环磷酸腺苷/环磷酸腺苷反应元件结合蛋白/脑源性神经营养因子信号通路 海马 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学] R749.4[医药卫生—临床医学]