AK-7对脑缺血大鼠杏仁核NLRP3蛋白表达的影响  

Effect of AK-7 on NLRP3 protein expression in amygdala of rats with cerebral ischemia

作　　者：罗潇潇 郭忻然 汪琴琴 赵健[2] 缪化春[2] 丁见[2] 郑兰荣[1] Luo Xiaoxiao;Guo Xinran;Wang Qinqin;Zhao Jian;Miao Huachun;Ding Jian;Zheng Lanrong(Developmental biology laboratory&Cell electrophysiology laboratory;Teaching and research room of human anatomy,Wannan Medical College,Wuhu,Anhui,241000,China)

机构地区：[1]皖南医学院发育生物学研究室、皖南医学院细胞电生理学研究室,安徽芜湖241000 [2]皖南医学院人体解剖学教研室,安徽芜湖241000

出　　处：《齐齐哈尔医学院学报》2022年第20期1906-1911,共6页Journal of Qiqihar Medical University

基　　金：皖南医学院校重点项目科研基金(WK2019Z05);皖南医学院2020年度中青年科研基金(WK202021)。

摘　　要：目的研究SIRT2(沉默信息调节因子2相关酶2)的特异性抑制剂AK-7对脑缺血大鼠杏仁核NLRP3(核苷酸结合寡聚域样受体蛋白3)蛋白表达的影响,初步探讨其作用机制。方法将36只成年雄性SPF级SD大鼠,适应性饲养1周后,随机分为假手术组、模型组和抑制剂组共三组,每组各12只。模型组给予栓塞大鼠右侧大脑中动脉法制造局灶性脑缺血的模型。造模成功后,抑制剂组大鼠分别在造模前30分钟、造模后6小时、24小时腹腔注射AK-7(20 mg/kg),假手术组及模型组大鼠在上述时间点腹腔注射同等体积的生理盐水。采用Longa评分于造模后第1、3、7天评估各组大鼠的神经功能缺损情况,通过尼氏染色法检测各组大鼠杏仁核内神经元形态变化;免疫组织化学方法和Western bolt法检测各组大鼠杏仁核SIRT2、NLRP3表达情况;ELISA法检测各组大鼠动脉血液中炎症因子IL-1β、IL-18的含量。结果与假手术组相比较,模型组与抑制剂组大鼠神经功能评分均显著增加(P<0.0001);尼氏染色结果显示模型组杏仁核内神经元细胞明显皱缩,细胞排列不规则,抑制剂组神经元形态与假手术组趋近;免疫组织化学染色和Western bolt结果显示模型组杏仁核SIRT2、NLRP3蛋白表达显著增加(P<0.05);ELISA法显示炎性因子IL-1β、IL-18含量显著增加。与模型组相比较,抑制剂组在脑缺血第7天时神经功能评分显著降低(P<0.01),免疫组织化学染色和Western bolt结果显示杏仁核SIRT2、NLRP3蛋白表达显著降低(P<0.05);ELISA法显示炎性因子IL-1β、IL-18含量显著降低。结论抑制SIRT2表达可对短期脑缺血大鼠起到神经保护作用,其机制可能与抑制NLRP3及其下游炎症因子IL-1β、IL-18的表达有关。Objective To study the effect of AK-7,a specific inhibitor of Silent information regulator 2 related enzyme 2(SIRT2),on nod-like receptor protein 3(NLRP3)protein expression in nucleotide binding oligomerization domainin the amygdala of rats with cerebral ischemia,and to explore its mechanism preliminarily.Methods Thirty-six adult male SPF SD rats were randomly divided into sham operation group,model group and inhibitor group after adaptive feeding for one week,with twelve rats in each group.The cerebral focal ischemia model was established via embolize the right middle cerebral artery.After modeling,the rats in inhibitor group were intraperitoneally injected with AK-7(20mg/kg)at 30min before modeling,6h and 24h after modeling,respectively.The rats in sham operation group and model group were intraperitoneally injected with the same volume of normal saline at the above time points.The Longa score was used to evaluate the neurological deficits of rats in each group on day 1,3,and 7 after modeling,and Nissl staining was used to detect the morphological changes of neurons in the amygdala of rats in each group.Immunohistochemistry and Western bolt were used to detect the expression of SIRT2 and NLRP3 in amygdala of each group.The levels of inflammatory cytokines IL-1βand IL-18 in arterial blood were detected by ELISA.Results Compared with the sham operation group,the neural function scores of the model group and the inhibitor group were increased(P<0.0001).The neuronal cells in the amygdala of the model group were significantly shrunk and irregularly arranged.The morphology of neurons in the inhibitor group was similar to that in the sham operation group.The protein expressions of SIRT2 and NLRP3 in amygdala of model group were significantly increased(P<0.05).The content of inflammatory factors IL-1βand IL-18 increased significantly.Compared with the model group,the inhibitor group had significantly lower neurological function scores on day 7 of cerebral ischemia,the protein expressions of SIRT2 and NLRP3 in amygdal

关 键 词：脑缺血 杏仁核 SIRT2 NLRP3 AK-7 

分 类 号：R743.3[医药卫生—神经病学与精神病学]