胱天蛋白酶募集域蛋白9在中重度缺血性脑卒中发病早期阶段的作用  

The role of caspase recruitment domain-containing protein 9 in the early stage of severe ischemic brain injury

作　　者：黄坚友 黄亮[1] 丁建文 曹春水[1] 熊华威[1] 李洋[1] Huang Jian-you;Huang Liang;Ding Jian-wen;Cao Chun-shui;Xiong Hua-wei;Li Yang(Emergency Department,the First Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第一附属医院急诊科,江西南昌330006

出　　处：《中国急救医学》2021年第4期319-323,共5页Chinese Journal of Critical Care Medicine

基　　金：北京协和医院睿E急诊医学研究专项基金(2018012)。

摘　　要：目的观察胱天蛋白酶募集域蛋白9(CARD9)在急性中重度缺血性脑卒中发病早期阶段的作用,探讨脑卒中炎症的发病机制。方法入选36例美国国立卫生研究院卒中量表(NIHSS)评分> 5分的急性中度及以上缺血性脑卒中患者为病例组;入选20例年龄、高血压、糖尿病等基础病与病例组差异无统计学意义的志愿者作为对照组。采集病例组发病第1、3、7天及对照组第1天外周血单个核细胞(PBMC)。Western blot检测CARD9表达水平及核转录因子-κB(NF-κB)活性(p-p65/p65水平);RT-PCR检测炎症因子肿瘤坏死因子-ɑ(TNF-ɑ)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6) mRNA的表达水平;病例组第1、3、7天使用NIHSS评分评估脑缺血损伤程度。结果 Western blot结果表明,入院第1、3、7天时,病例组CARD9表达及NF-κB活性与对照组比较显著升高(CARD9:1.17±0.14、1.08±0.15、0.97±0.16 vs. 0.34±0.02,NF-κB:1.78±0.06、1.70±0.07、1.46±0.04 vs. 1.21±0.12,P均<0.05);RT-PCR结果表明,入院第1、3、7天时,病例组TNF-αmRNA表达与对照组比较显著升高(5.34±0.14、4.57±0.15、3.58±0.19 vs. 1.00±0.08,P均<0.01),病例组IL-1βmRNA表达与对照组比较显著升高(6.49±0.12、4.85±0.23、3.38±0.31 vs. 1.00±0.11,P均<0.01),病例组IL-6 mRNA表达与对照组比较显著升高(10.29±0.55、7.32±0.44、4.56±0.33 vs. 1.00±0.07,P均<0.01)。Pearson相关分析表明,脑卒中患者PBMC中NF-κB活化、TNF-α、IL-1β、IL-6 mRNA及NIHSS评分与CARD9表达呈正相关(r值分别为0.971、0.974、0.970、0.982、0.891,P均<0.01)。结论 CARD9在早期中重度缺血性脑卒中患者PBMC中的表达增高,可能通过NF-κB信号通路介导缺血性脑损伤炎症反应。Objective To observe the role of caspase recruitment domain-containing protein 9( CARD9) in the early stage of acute moderate to severe ischemic stroke and to explore the pathogenesis of stroke inflammation. Methods Peripheral blood monocytes( PBMC) were collected from 36 patients of acute moderate to severe ischemic stroke with NIHSS score more than 5 on 1 st,3 rd and 7 th day.Twenty volunteers with no significant difference in age,hypertension and diabetes compared with case group were selected as control group. CARD9 expression and nuclear factor-κB( NF-κB) activity( p-p65/p65) in PBMC were detected by Western blot. RT-PCR was used to detect PBMC inflammatory factor TNF-ɑ,IL-lβ,IL-6 mRNA expression level,NIHSS score was used to evaluate the degree of cerebral ischemia injury in the case group on 1 st,3 rd and 7 th day. Results Western blot results showed that on 1 st,3 rd and 7 th day after admission,CARD9 expression and NF-κB activity in the case group were significantly higher than those in the control group( CARD9: 1. 17 ± 0. 14,1. 08 ±0. 15,0. 97 ± 0. 16 vs. 0. 34 ± 0. 02,NF-κB: 1. 78 ± 0. 06,1. 70 ± 0. 07,1. 46 ± 0. 04 vs. 1. 21 ±0. 12,P< 0. 05). RT-PCR results showed that on 1 st,3 rd and 7 th day after admission,TNF-ɑmRNA expression was significantly higher in the case group than in the control group( 5. 34 ± 0. 14,4. 57 ± 0. 15,3. 58 ± 0. 19 vs. 1. 00 ± 0. 08,P< 0. 01). The expression of IL-1β mRNA was significantly higher in the case group than in the control group( 6. 49 ± 0. 12,4. 85 ± 0. 23,3. 38 ± 0. 31 vs. 1. 00 ± 0. 11,P< 0. 01). The expression of IL-6 mRNA was significantly higher in the case group than in the control group( 10. 29 ± 0. 55,7. 32 ± 0. 44,4. 56 ± 0. 33 vs. 1. 00 ± 0. 07,P< 0. 01).Pearson correlation analysis showed that NF-κB activation,TNF-ɑ,IL-1β,IL-6 mRNA in PBMC and NIHSS score were positively related to CARD9 in the patients with cerebral stroke( r = 0. 971,0. 974,0. 970,0. 982,0. 891,P< 0. 01). Conclusions The expressions of CARD9 in PBMC of t

关 键 词：胱天蛋白酶募集域蛋白9(CARD9) 核转录因子-κB(NF-κB) 炎症因子 急性缺血性脑卒中(AIS) 外周血单个核细胞(PBMC) 

分 类 号：R743.3[医药卫生—神经病学与精神病学]