光生物调节促进慢性低灌注大鼠海马神经元再生改善认知功能及其抗炎症作用  

Photobiomodulation Promotes Hippocampal Neurogenesis and Improves Cognitive Function and Anti-Inflammatory Injury in Rats With Chronic Cerebral Hypoperfusion

作　　者：涂静宜[1] 黄媛媛 黄莺 吴蒙[1] 王瑞敏 TU Jingyi;HUANG Yuanyuan;HUANG Ying;WU Meng;WANG Ruimin(Pathological Teaching and Research Division,Department of Basic Medicine,Tangshan Vocational and Technical College,Tangshan 063000,China;Neurobiology Institute,School of Public Health,North China University of Science and Technology,Tangshan 063210,China)

机构地区：[1]唐山职业技术学院基础医学部病理教研室,唐山063000 [2]华北理工大学公共卫生学院神经生物学研究所,唐山063210

出　　处：《四川大学学报（医学版）》2023年第5期965-971,共7页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金面上项目(No.31171354);河北省人力资源和社会保障厅择优资助基金(No.A202002028)资助。

摘　　要：目的探讨光生物调节(photobiomodulation,PBM)对脑慢性低灌注大鼠海马神经元再生及认知功能和炎症损伤的影响。方法将雌性SD大鼠去双侧卵巢,1周后随机分为假手术组(Sham)、BCCAO组、PBM干预组(BCCAO+PBM),每组8只。BCCAO组永久性结扎大鼠双侧颈总动脉(bilateral common carotid artery occlusion,BCCAO)建立脑慢性低灌注模型,但不予光照;Sham组只钝性分离左、右侧颈总动脉,但不结扎,不予光照。BCCAO+PBM组在永久性结扎BCCAO的基础上,采用808 nm激光隔日照射大脑额叶皮层1个月,每次5 min,光剂量为20 mW/cm^(2)。造模完成后第86~90天行Morris水迷宫测试观察大鼠的空间学习记忆功能,第90天处死大鼠进行免疫荧光染色和Western blot分析。免疫荧光染色检测海马齿状回(dentate gyrus,DG)区细胞增殖标记物5-溴脱氧尿嘧啶核苷(BrdU)、星形胶质细胞标记物胶质纤维酸性蛋白(GFAP)、新生神经元前体细胞的特异性标志蛋白DCX、成熟神经元标志蛋白NeuN以及小胶质细胞标志蛋白Iba1的表达情况。Western blot分析海马区Iba1,炎症小体组成NLRP3、ASC、cleaved caspase-1的蛋白水平。结果水迷宫潜伏实验中,与BCCAO组相比,PBM干预组大鼠找到水下平台的时长缩短;在探索实验中,移除平台后大鼠在原平台象限探索的时间增加(P<0.05)。与BCCAO组相比,PBM干预组GFAP、Iba1的免疫荧光强度减弱(P<0.01);与Sham组和BCCAO组相比,PBM干预组可增加海马DG区BrdU阳性细胞的数量(P<0.05);3组间NeuN阳性细胞数差异无统计学意义,而PBM干预组DCX阳性细胞数增加(P<0.001),且DCX~+/NeuN~+共定位的细胞数较BCCAO组增多(P<0.001)。Western blot结果显示,与BCCAO组相比,PBM干预组Iba1、NLRP3、cleaved caspase-1蛋白的表达量均降低(P<0.05),ASC蛋白的表达量差异无统计学意义。结论PBM能有效改善慢性低灌注大鼠的空间学习记忆功能,抑制胶质细胞的激活、降低NLRP3炎症小体介导的炎症损�Objective To investigate the effect of photobiomodulation(PBM)on hippocampal neurogenesis,cognitive function,and inflammatory injury in rats with chronic cerebral hypoperfusion.Methods Bilateral ovariectomy(OVX)was performed on female Sprague-Dawley(SD)rats.One week later,the rats were randomly assigned to three groups,Sham surgery(or Sham)group,bilateral common carotid artery occlusion(BCCAO)group,and PBM intervention(or BCCAO+PBM)group.There were 8 rats in each group.In the BCCAO group,chronic cerebral hyporeperfusion was induced by permanent ligation of bilateral common carotid arteries and no PBM was given.Rats in the Sham group underwent the same surgical procedure except for the occlusion of the two carotids arteries and no PBM was given.In addition to the BCCAO surgery,rats in the BCCAO+PBM group received 808 nm laser therapy(5 min each time at a laser dose of 20 mW/cm^(2))of the frontal cortex every other day for 1 month.Between 86 and 90 days after BCCAO,Morris water maze(MWM)was used to observe the spatial learning and memory function of the rats.The rats were sacrificed on day 90 and immunofluorescence staining and Western blot were performed thereafter.Immunofluorescence staining was used to determine the expression of 5-bromodeoxyuracil nucleoside(BrdU),a cell proliferation marker,glial fibrillary acidic protein(GFAP),an astrocyte marker,doublecortin(DCX),a specific marker of newborn neuron precursor cells,NeuN,a marker of mature neurons,and Iba1,a microglia marker,in the hippocampal dentate gyrus(DG)region.Western blot was performed to analyze the protein expressions of inflammasome components,NLRP3,ASC,cleaved caspase-1,and Iba1 in the hippocampus.Results In the latency trial of MWM test,BCCAO+PBM rats spent shorter periods of time finding the underwater platform than the BCCAO rats did.In the probe trial,after the platform that was original placed in a quadrant was removed,the BCCAO+PBM rats spent longer periods of time exploring the quadrant than the BCCAO animals did(P<0.05).Compared with BCCAO

关 键 词：脑慢性低灌注 光生物调节 神经元再生 神经炎症 学习记忆 

分 类 号：R741[医药卫生—神经病学与精神病学]