apelin受体及其同源二聚体促进血管性痴呆大鼠内源性神经干细胞增殖的研究  

Effects of apelin receptor and its homodimer on proliferation of endogenous neural stem cells in vascular dementia rats

作　　者：王德秀 李建设 殷月 高菁婕 王咏婧 蔡欣 王玉良[1] WANG De-xiu;LI Jian-she;YIN Yue;GAO Jing-jie;WANG Yong-jing;CAIXin;WANG Yu-liang(School of Basic Medicine,Weifang Medical University,Weifang 261053,China;School of Clinical Medicine,Weifang Medical University,Weifang 261053,China;School of Life Science and Technology,Weifang Medical University,Weifang 261053,China)

机构地区：[1]潍坊医学院基础医学院,山东潍坊261053 [2]潍坊医学院临床医学院,山东潍坊261053 [3]潍坊医学院生命科学与技术学院,山东潍坊261053

出　　处：《中国病理生理杂志》2022年第12期2147-2153,共7页Chinese Journal of Pathophysiology

基　　金：山东省中医药科技项目(No.2021Q071);山东省医药卫生科技发展计划项目(No.202102080543);山东省自然科学基金资助项目(No.ZR2022MC060);潍坊市科技发展计划项目(No.2021YX044)。

摘　　要：目的:探究apelin受体APJ及其同源二聚体对血管性痴呆(VD)大鼠内源性神经干细胞(NSCs)增殖的影响。方法:将48只8周龄清洁级雄性SD大鼠随机分为假手术组、模型组、apelin-13组(APJ同源二聚体组)和apelin-13+TM1(APJ的跨膜区1)组(APJ单体组),每组12只。采用双侧夹闭颈总动脉并腹腔注射硝普钠溶液的方法制备VD模型,apelin-13组和apelin-13+TM1组大鼠于造模后7 d通过侧脑室注射的方法给药,同时给予假手术组和模型组大鼠等体积生理盐水。模型制备后15 d,利用Morris水迷宫实验检测大鼠的学习记忆能力;HE染色观察大鼠海马CA1区锥体神经元的形态及数目;免疫荧光染色观察大鼠海马齿状回(DG)内源性NSCs的增殖情况;Western blot法测定大鼠海马组织增殖细胞核抗原(PCNA)、巢蛋白(nestin)和cleaved caspase-3的蛋白水平。结果:Morris水迷宫实验结果显示,模型组大鼠逃逸潜伏期(EL)较假手术组显著延长(P<0.01);而apelin-13组和apelin-13+TM1组大鼠的EL较模型组均显著缩短(P<0.05),且apelin-13组短于apelin-13+TM1组。HE染色结果显示,模型组锥体神经元数目较假手术组显著减少(P<0.01);与模型组相比,apelin-13和apelin-13+TM1组中锥体神经元损伤程度较轻,存活的神经元数目较多(P<0.01),且apelin-13组多于apelin-13+TM1组(P<0.01)。免疫荧光结果显示,模型组、apelin-13组和apelin-13+TM1组大鼠海马DG区均存在PCNA+nestin+DAPI+细胞。Western blot结果显示,与假手术组相比,模型组大鼠海马区nestin、PCNA和cleaved caspase-3的蛋白水平均显著升高(P<0.01),apelin-13组和apelin-13+TM1组的nestin和PCNA表达均高于模型组(P<0.01),而cleaved caspase-3的水平显著降低(P<0.01)。结论:APJ单体及其同源二聚体均能促进VD大鼠海马DG区NSCs的增殖,延缓海马CA1区锥体神经元的凋亡,并改善VD大鼠的认知能力,且APJ同源二聚体的效果优于单体。AIM:To investigate the effect of apelin receptor(APJ)and its homodimer on the proliferation of endogenous neural stem cells(NSCs)in rats with vascular dementia(VD).METHODS:Forty-eight male Sprague-Dawley rats aged 8 weeks were randomly divided into sham group,model group,apelin-13 group(APJ homodimer group)and apelin-13+transmembrane domain 1(TM1)group(APJ monomer group),with 12 rats in each group.The VD model was established by clamping the bilateral common carotid arteries and injecting sodium nitroprusside solution intraperitoneally.The rats in apelin-13 and apelin-13+TM1 groups were injected with drugs into the lateral ventricle at 7 d after modeling,while the rats in sham and model groups were given the same volume of normal saline.Morris water maze test was used to detect the learning and memory ability of rats.Hematoxylin-eosin(HE)staining was used to observe the morphology and number of pyramidal neurons in hippocampal CA1 area of rats.Immunofluorescence(IF)staining was used to detect the proliferation of endogenous NSCs in rat hippocampal dentate gyrus(DG).Western blot was used to detect the levels of proliferating cell nuclear antigen(PCNA),nestin and cleaved caspase-3 in rat hippocampal tissues at 15 d after modeling.RESULTS:Morris water maze test showed that the escape latency(EL)of the rats in model group was significantly longer than that in sham group(P<0.01),the EL in apelin-13 and apelin-13+TM1 groups was shorter than that in model group(P<0.05),and that in apelin-13 group was shorter than that in apelin-13+TM1 group.The results of HE staining showed that the number of pyramidal neurons in model group was significantly less than that in sham group(P<0.01).Compared with model group,the injury of pyramidal neurons in apelin-13 and apelin-13+TM1 groups was attenuated,and the number of surviving neurons was increased(P<0.01).Moreover,the number of pyramidal neurons in apelin-13 group was larger than that in apelin-13+TM1 group(P<0.01).The IF staining results showed that there were PCNA+nestin+DAPI+ce

关 键 词：血管性痴呆 Apelin受体 同源二聚体 神经干细胞 细胞增殖 

分 类 号：R742[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]