下丘脑外侧区星形胶质细胞NLRP3在小鼠失血性休克复苏后焦虑样行为中的作用  

Role of astrocytic NLRP3 in lateral hypothalamus in anxiety-like behaviors after hemorrhagic shock and resuscitation in mice

作　　者：张冬雪[1] 苗慧涛 宋荣欣 李汶广 贾世艳 Zhang Dongxue;Miao Huitao;Song Rongxin;Li Wenguang;Jia Shiyan(Department of Gerontology,Cangzhou Central Hospital,Cangzhou 061000,China;Hebei Key Laboratory of Integrated Traditional and Western Medicine in Osteoarthrosis Research,Cangzhou 061000,China;Department of Anesthesiology,Hebei Province Cangzhou Hospital of Integrated Traditional and Western Medicine,Cangzhou 061000,China;Graduate School,Hebei Medical University,Shijiazhuang 050011,China)

机构地区：[1]沧州市中心医院老年医学科,沧州061000 [2]河北省中西医结合骨关节病重点实验室,沧州061000 [3]河北省沧州中西医结合医院麻醉科,沧州061000 [4]河北医科大学研究生院,石家庄050011

出　　处：《中华麻醉学杂志》2023年第8期981-985,共5页Chinese Journal of Anesthesiology

基　　金：河北省自然科学基金青年科学基金 (H2021110004)。

摘　　要：目的:评价下丘脑外侧区星形胶质细胞NOD样受体相关蛋白3(NLRP3)在小鼠失血性休克复苏后焦虑样行为中的作用。方法:清洁级雄性C57BL/6小鼠48只,10周龄,体质量25~30 g,采用随机数字表法分为4组( n=12):假手术组(C组)、失血性休克复苏组(H组)、失血性休克复苏+腺病毒组(HI组)和失血性休克复苏+对照腺病毒组(HIV组)。H组、HI组和HIV组小鼠通过股静脉放血-回输法建立失血性休克复苏模型;建模前21 d时,HI组小鼠在立体定位仪引导下向双侧下丘脑外侧区注射AAV-GfaABC1D-EGFP-Cre腺病毒,HIV组注射AAV-GfaABC1D-EGFP对照腺病毒。复苏后14 d时,采用埋珠实验和高架十字迷宫实验评估焦虑样行为;行为学实验结束后立即处死小鼠,取含有下丘脑外侧区的脑组织,采用免疫荧光染色法,测定紫藤凝集素荧光强度反映细胞外基质表达,测定NLRP3和胶质纤维酸性蛋白(GFAP)共定位情况,计算cleaved caspase-1/GFAP、IL-18/GFAP阳性细胞数占总细胞的百分比。 结果:与C组比较,H组、HI组和HIV组埋珠数量减少,开放臂停留时间百分比降低,下丘脑外侧区细胞外基质表达下调,cleaved caspase-1/GFAP和IL-18/GFAP阳性细胞百分比增加,HI组NLRP3与GFAP共定位系数降低( P<0.01)。与H组比较,HI组埋珠数量增加,开放臂停留时间百分比降低,下丘脑外侧区细胞外基质表达上调,NLRP3与GFAP共定位系数降低,cleaved caspase-1/GFAP和IL-18/GFAP阳性细胞百分比降低( P<0.01),HIV组上述指标差异无统计学意义( P>0.05)。 结论:小鼠失血性休克复苏后焦虑样行为与下丘脑外侧区星形胶质细胞NLRP3诱发焦亡,减少细胞外基质有关。Objective To evaluate the role of astrocytic NOD-like receptor protein 3(NLRP3)in the lateral hypothalamus(LHA)in anxiety-like behaviors after hemorrhagic shock and resuscitation in mice.Methods Forty-eight clean-grade male C57BL/6 mice,aged 10 weeks,weighing 25-30 g,were divided into 4 groups(n=12 each)using a random number table method:sham operation group(group C),hemorrhagic shock and resuscitation group(group H),hemorrhagic shock and resuscitation+adeno-associated virus group(group HI),and hemorrhagic shock and resuscitation+control virus group(group HIV).The model of hemorrhagic shock and resuscitation was developed by bleeding and re-transfusion through the femoral vein in H,HI and HIV groups.At 21 days before developing the model,AAV-GfaABC1D-EGFP-Cre was injected into bilateral LHA in group HI,and AAV-GfaABC1D-EGFP was administered as a control in group HIV.Anxiety-like behaviors were evaluated by EPM-maze and bead-burying tests at 14 days after resuscitation.Mice were immediately sacrificed at the end of behavioral tests,and LHA-containing brain tissues were obtained for determination of co-localization of NLRP3 with glial fibrillary acidic protein(GFAP),the fluorescence intensity of Wisteria floribunda agglutinin was measured using immunofluorescent staining to reflect the expression of extracellular matrix in the LHA,and the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells in total cells was calculated.Results Compared with group C,the number of buried beads and percentage of time of staying at the open arm were significantly decreased,the expression of extracellular matrix in the LHA was down-regulated,and the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells was increased in H,HI and HIV groups,and the co-localization coefficient of NLRP3 and GFAP was significantly decreased in group HI(P<0.01).Compared with group H,the number of buried particles and percentage of time of staying at the open arm were significantly decreased,the expression of extracellular matrix in

关 键 词：休克 出血性 复苏术 焦虑 下丘脑区 侧 星形细胞 NLR家族 热蛋白结构域包含蛋白3 

分 类 号：R459.7[医药卫生—急诊医学]