摘要
目的研究NLRP3炎症小体信号通路相关分子在缺氧缺血性脑损伤(hypoxic-ischemic brain damage,HIBD)新生大鼠海马中的表达情况,初步探讨NLRP3炎症小体信号通路在HIBD新生大鼠脑损伤中的作用。方法50只7日龄新生SD大鼠随机分为假手术组(Sham组,20只)和模型组(HIBD组,30只)。参照改良Rice法建立HIBD动物模型。于术后不同时间点取脑组织,Western Blot检测左侧海马组织中NLRP3、ASC、pro-Caspase-1、Caspase-1、GSDMD、IL-1β、IL-18蛋白表达情况;HE染色和尼氏染色观察左侧海马DG区脑组织病理变化及神经元损伤情况;免疫组织化学染色检测左侧海马DG区NLRP3、ASC、Caspase-1蛋白表达情况;RT-qPCR检测左侧海马组织中NLRP3、ASC、Caspase-1、GSDMD、IL-β、IL-18 mRNA表达情况。结果与Sham组相比,HIBD组术后24 h、48 h NLRP3蛋白表达水平升高;术后72 h ASC及Caspase-1蛋白表达水平升高;术后各时间点pro-Caspase-1蛋白表达水平无明显变化。术后72 h,与Sham组相比,HIBD组HE染色、尼式染色、免疫组织化学染色、Western Blot及RT-qPCR结果分别显示,左侧海马DG区细胞层数相对减少、细胞排列相对不规则;神经元数目丢失增多;NLRP3、ASC、Caspase-1蛋白表达增强;NLRP3、ASC、Caspase-1、GSDMD、IL-1β、IL-18 mRNA及蛋白表达水平升高,差异具有统计学意义(P<0.05)。结论NLRP3炎症小体信号通路在HIBD新生大鼠海马组织中表达增高,可能导致HIBD新生大鼠神经细胞死亡,未来可能成为新生儿HIBD的潜在治疗靶点。
Objective To investigate the expression of NLRP3 inflammatory signaling pathway-related molecules in hippocampus of neonatal rats with hypoxic-ischemic brain damage(HIBD),and to explore the role of NLRP3 inflammatory signaling pathway in hippocampal injury of neonatal rats with HIBD. Methods Fifty 7-dayold neonatal SD rats were randomly divided into sham group(20 rats)and model group(HIBD group,30 rats).The animal model of HIBD was established according to the modified Rice method. The expression of NLRP3,ASC,pro-Caspase-1,Caspase-1,GSDMD,IL-1β and IL-18 in the left hippocampus was detected by Western Blot. The pathological changes and neuronal damage in the left hippocampus DG were observed by HE staining and Nissl staining. The expression of NLRP3,ASC and Caspase-1 in the left hippocampus DG was detected by immunohistochemical staining. RT-qPCR was used to detect the expression of NLRP3,ASC,Caspase-1,GSDMD,IL-1βand IL-18 in left hippocampus. Results Compared with Sham group,the expression of NLRP3 protein were increased in 24 hours and 48 hours after surgery in HIBD group. ASC and Caspase-1 protein were increased in 72 hours after surgery in HIBD group. The expression of pro-Caspase-1 protein did not change significantly at all time points after operation. The number of cell layers in DG region of left hippocampus was relatively reduced and the cell arrangement was relatively irregular 72 hours after operation in HIBD group than Sham group. Moreover,the number of neurons,the expression of NLRP3,ASC,Caspase-1 protein as well as the expression of NLRP3,ASC,Caspase-1,GSDMD,IL-1β,IL-18 mRNA and protein surface were significantly increased in HIBD group than Sham group72 hours after operation(P < 0.05). Conclusion The increased expression of NLRP3 inflammatory signaling pathway in hippocampus of neonatal HIBD rats may lead to the death of neurons in neonatal HIBD rats,which may become a potential therapeutic target for neonatal HIBD in the future.
作者
胡兴
苟知贤
王幽梦
黄林
周月
鲁利群
HU Xing;GOU Zhixian;WANG Youmeng;HUANG Lin;ZHOU Yue;LU Liqun(The First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China)
出处
《实用医学杂志》
CAS
北大核心
2019年第24期3746-3751,共6页
The Journal of Practical Medicine
基金
四川省科技厅应用基础研究项目(编号：2018JY 0301)
