摘要
目的评价长期摄入ω-3多不饱和脂肪酸(ω-3 PUFAs)对术后认知功能障碍(POCD)小鼠海马小胶质细胞活化的影响。方法清洁级健康雄性C57BL/6小鼠96只,8周龄,体重18~24 g,按照体重分层,采用随机数字表法分为4组(n=24):对照饲料组(C组)、ω-3 PUFAs组(ω组)、对照饲料+POCD组(C+P组)和ω-3 PUFAs+POCD组(ω+P组)。ω组和ω+P组给予特制ω-3 PUFAs饲料[二十二碳六烯酸(DHA)0.14 g/100 g,二十碳五烯酸(EPA)0.03 g/100 g]喂养12周,C组和C+P组给予对照饲料喂养12周。12周喂养结束后,采用异氟烷麻醉下行胫骨骨折手术的方法制备小鼠POCD模型。造模后第1和3天进行条件恐惧实验和Y迷宫实验。在行为学测试结束后处死小鼠取海马组织,采用气相色谱-质谱联用法测定DHA和EPA含量,免疫荧光染色法测定Iba-1阳性小胶质细胞密度,Western blot法检测成熟脑源性神经营养因子(mBDNF)和前体脑源性神经营养因子(pro-BDNF)的表达,ELISA法测定IL-1β和IL-6含量。结果与C组比较,ω组海马DHA和EPA含量升高,场景恐惧测试僵直时间百分比增加,mBDNF/pro-BDNF比值升高(P<0.05),轮替正确率、海马Iba-1阳性小胶质细胞密度、IL-1β和IL-6含量差异无统计学意义(P>0.05),C+P组小鼠DHA与EPA含量差异无统计学意义(P>0.05),术后第1和3天场景恐惧测试僵直时间百分比和轮替正确率减少,海马Iba-1阳性小胶质细胞密度增加,IL-1β和IL-6含量升高,mBDNF/pro-BDNF比值降低(P<0.05);与C+P组比较,ω+P组海马DHA和EPA含量升高,术后第1和3天场景恐惧测试僵直时间百分比和轮替正确率升高,海马Iba-1阳性小胶质细胞密度减少,海马IL-1β和IL-6含量降低,mBDNF/pro-BDNF比值增加(P<0.05)。结论长期摄入ω-3 PUFAs可改善POCD小鼠认知功能,其机制可能与抑制海马小胶质细胞活化,减轻炎症反应,从而升高mBDNF/pro-BDNF比值有关。
Objective To evaluate the effect of long-term intake ofω-3 polyunsaturated fatty acids(ω-3 PUFAs)on the activation of hippocampal microglia in a mouse model of postoperative cognitive dysfunction(POCD).Methods Ninety-six clean-grade healthy male C57BL/6 mice,aged 8 weeks,weighing 18-24 g,were stratified according to body weight and divided into 4 groups(n=24 each)by a random number table method:control diet group(group C),ω-3 PUFAs group(groupω),control diet plus POCD group(group C+P)andω-3 PUFAs plus POCD group(groupω+P).Mice were fed a specialω-3 PUFAs diet(DHA 0.14 g/100 g,EPA 0.03 g/100 g)for 12 weeks in groupωand groupω+P,while mice were fed with a control diet for 12 weeks in group C and group C+P.Tibial fracture procedures were performed under isoflurane anesthesia to develop the POCD model after 12 weeks of feeding.The fear conditioning test and Y maze test were performed on 1st and 3rd days after developing the model.The mice were sacrificed after behavioral tests,and the hippocampal tissues were removed for determination of the contents of docosahexaenoic acid(DHA)and eicosapentaenoic acid(EPA)(by gas chromatography-mass spectroscopy),density of Iba-1 positive microglia(by immunofluorescence staining),and expression of mature brain-derived neurotrophic factor(mBDNF)and precursor brain-derived neurotrophic factor(pro-BDNF)(by Western blot),and contents of interleukin-1beta(IL-1β)and interleukin-6(IL-6)(by enzyme-linked immunosorbent assay).Results Compared with group C,the contents of DHA and EPA were significantly increased,the percentage of freezing time in the contextual test was increased,mBDNF/pro-BDNF ratio was increased(P<0.05),no significant change was found in the rotation accuracy in Y maze test,density of Iba-1 positive microglia and contents of IL-1βand IL-6 in hippocampus(P>0.05)in groupω(P<0.05),and no significant change was found in the contents of DHA and EPA(P>0.05),the percentage of freezing time in the contextual test and accuracy of rotation in Y maze test were decreased o
作者
韩玉强
董瑞
刘帅
张富俊
王子沫
夏天娇
顾小萍
Han Yuqiang;Dong Rui;Liu Shuai;Zhang Fujun;Wang Zimo;Xia Tianjiao;Gu Xiaoping(Nanjing Drum Tower Hospital Clinical College of Xuzhou Medical University,Nanjing 210009,China;Department of Anesthesiology,Affiliated Drum Tower Hospital,Medical School of Nanjing University,Nanjing 210008,China;Medical School,Nanjing University,Nanjing 210093,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2022年第1期44-49,共6页
Chinese Journal of Anesthesiology
基金
国家自然科学基金(81730033,82001132,82171193)
十三五强卫工程-重点人才项目(ZDRCA2016069)
南京大学医学院研究生科研与实践创新计划项目(2021)。
