鱼藤酮对大鼠认知功能及其海马Warburg效应和小胶质细胞极化的影响  

Effects of Rotenone on Cognitive Function as well as Hippocampal Warburg Effect and Microglia Polarization in Rats

作　　者：胡宇 唐慧玲 田清 张平[4] 邹伟[4] 唐小卿 HU Yu;TANG Hui-ling;TIAN Qing;ZHANG Ping;ZOU Wei;TANG Xiao-qing(Institute of Clinical Laboratory,The First Affiliated Hospital,University of South China,Hengyang 421001,China;Institute of Neurology,The First Affiliated Hospital,University of South China,Hengyang 421001,China;Institute of Neuroscience,Hengyang Medical College,University of South China,Hengyang 421001,China;Department of Neurology,The Affiliated Nanhua Hospital,University of South China,Hengyang 421001,China)

机构地区：[1]南华大学附属第一医院检验科,湖南衡阳421001 [2]南华大学附属第一医院神经疾病研究所,湖南衡阳421001 [3]南华大学衡阳医学院神经科学研究所,湖南衡阳421001 [4]南华大学附属南华医院神经病学教研室,湖南衡阳421001

出　　处：《中山大学学报（医学科学版）》2021年第6期801-807,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81771178);湖南省自然科学基金(2020JJ4553);湖南省临床医疗技术创新引导项目(2020SK51908)。

摘　　要：【目的】探究鱼藤酮(ROT)对大鼠认知功能及其海马Warburg效应和小胶质细胞极化的影响。【方法】雄性SD大鼠随机分为对照组和ROT处理组。ROT处理组大鼠颈背部皮下注射ROT(2 mg/kg),对照组注射等量的溶剂。Y迷宫实验和新物体识别实验评估大鼠的认知功能,Western blot检测海马Warburg效应相关蛋白己糖激酶2(HK2)、丙酮酸激酶同工酶2(PKM2)、丙酮酸脱氢酶激酶1(PDK-1)、丙酮酸脱氢酶(PDH)、乳酸脱氢酶A(LDHA)和小胶质细胞极化标志性蛋白诱导型一氧化氮合成酶(iNOS)、精氨酸酶1(Arg-1)、几丁质酶3样分子3(Ym-1)的表达水平,乳酸试剂盒检测海马组织乳酸含量,Griess法测定海马组织一氧化氮(NO)含量,酶联免疫吸附测定法检测海马组织IL-1β、IL-4和TGF-β1含量。【结果】ROT处理组大鼠在Y迷宫实验中的正确交替率(P<0.05)和在新物体识别实验中的识别指数(P<0.001)较对照组降低,表明ROT损害大鼠的认知功能。ROT处理组大鼠海马组织HK2(P<0.05)、PKM2(P<0.01)、PDK-1(P<0.05)和LDHA(P<0.05)的表达下降,PDH的表达上升(P<0.05),乳酸的含量减少(P<0.05),表明ROT抑制大鼠海马Warburg效应。ROT处理组大鼠海马组织iNOS的表达增加(P<0.05)、NO的含量增加(P<0.05)、IL-1β的分泌增加(P<0.01)、Arg-1(P<0.01)和Ym-1(P<0.01)的表达降低、IL-4(P<0.05)和TGF-β1(P<0.05)的分泌减少,表明ROT促进大鼠海马小胶质细胞M1极化。【结论】ROT可破坏大鼠的认知功能、抑制海马Warburg效应、促进海马小胶质细胞M1极化。【Objective】To investigate the effects of rotenone(ROT)on cognitive function of rats,as well as on Warburg effect and microglia polarization in the hippocampus of rats.【Methods】Male Sprague Dawley(SD)rats were randomly divided into ROT-treated group and control group which were subcutaneously injected with ROT(2 mg/kg)and same volume of saline respectively.The cognitive function of rats was evaluated by Y-maze test and novel object recognition(NOR)test.Western blot(WB)was conducted to detect the expression levels of Warburg effect related proteins including hexokinase 2(HK2),pyruvate kinase isozyme M2(PKM2),pyruvate dehydrogenase kinase 1(PDK-1),pyruvate dehydrogenase(PDH),and lactate dehydrogenase A(LDHA)in the hippocampus,as well as the expression levels of the microglia polarization marker proteins including inducible nitric oxide synthase(iNOS),arginase-1(Arg-1),and chitinase 3-like protein 3(Ym-1)in the hippocampus.The lactate production and concentration of nitric oxide(NO)in the hippocampus were determined by lactate assay kit and Griess method,respectively.Enzyme-linked immunosorbent assay was used to measure the levels of interleukin-1β(IL-1β),interleukin-4(IL-4),and transforming growth factorβ1(TGF-β1)in the hippocampus.【Results】The ROT-treated rats showed impaired cognitive function,presented as a significantly lower alternation rate(P<0.05)in Y-maze test and recognition index(P<0.001)in NOR test compared with control rats.In the hippocampus of ROT-treated rats,the expressions of HK2(P<0.05),PKM2(P<0.01),PDK-1(P<0.05)and LDHA(P<0.05)were downregulated,The PDH expression was upregulated(P<0.05),and the lactate production(P<0.05)was reduced,indicating that ROT inhibited Warburg effect in the hippocampus of rats.The hippocampus of ROT-treated rats also exhibited increased iNOS expression(P<0.05),NO concentration(P<0.05),and IL-1βlevel(P<0.01),decreased expression of Arg-1(P<0.01)and Ym-1(P<0.01),and reduced levels of IL-4(P<0.05)and TGF-β1(P<0.05),indicating that ROT promoted M1 polarizat

关 键 词：鱼藤酮 Warburg效应 小胶质细胞极化 认知障碍 帕金森病 

分 类 号：R741.02[医药卫生—神经病学与精神病学]