组蛋白去乙酰化酶3特异性抑制剂RGFP966通过抑制神经细胞焦亡对创伤性脑损伤产生神经保护作用  

Neuroprotective effect of histone deacetylase 3 specific inhibitor RGFP966 on traumatic brain injury by inhibiting pyroptosis

作　　者：徐兰娟 贾宝辉 安婷婷 吴琼 李向阳 马轶凡 丁慧慧 宋涛 李丽青 李成建 Xu Lanjuan;Jia Baohui;An Tingting;Wu Qiong;Li Xiangyang;Ma Yifan;Ding Huihui;Song Tao;Li Liqing;Li Chengjian(Department of Critical Care Medicine,Zhengzhou Central Hospital,Zhengzhou University,Zhengzhou 450001,China)

机构地区：[1]郑州大学附属郑州中心医院重症医学科,郑州450001

出　　处：《中华神经医学杂志》2023年第9期875-883,共9页Chinese Journal of Neuromedicine

基　　金：2021年度河南省科技攻关项目(212102310673);2021年度河南省高等学校重点科研项目计划(21A320056)。

摘　　要：目的探讨组蛋白去乙酰化酶3(HDAC3)特异性抑制剂RGFP966对创伤性脑损伤(TBI)大鼠早期脑损伤的神经保护作用及其机制。方法将48只SD大鼠按随机数字表法分为假手术组、TBI组、TBI+溶媒对照组、TBI+RGFP966组,每组12只。后3组大鼠采用液压冲击脑损伤法建立TBI模型,其中TBI+RGFP966组于造模后30 min经腹腔注射RGFP966(溶于1%DMSO中,剂量为10 mg/kg,2次/d,共给药3 d),TBI+溶媒对照组同期注射等量DMSO。于造模后第3天,分别应用改良神经功能缺损评分(mNSS)评估各组大鼠的神经功能,干湿重法检测各组大鼠损伤侧大脑皮层含水量,尼氏染色检测各组大鼠损伤侧额叶皮层组织中损伤神经元比例,免疫组化染色及Western blotting检测各组大鼠损伤侧额叶皮层组织中HDAC3及焦亡相关蛋白表达,ELISA法检测各组大鼠血清中炎性因子含量。结果造模后第3天,与TBI+溶媒对照组比较,TBI+RGFP966组大鼠mNSS评分[(9.83±0.75)分vs.(6.67±0.82)分]、脑组织含水量[(82.73±0.36)%vs.(80.92±0.66)%]明显下降,损伤神经元比例[(75.60±7.44)%vs.(55.87±4.10)%]明显下降,HDAC3蛋白表达明显下降(0.67±0.09 vs.0.51±0.07),乙酰化H3、乙酰化H4蛋白表达明显上升(0.81±0.02 vs.1.22±0.02;0.74±0.01 vs.1.07±0.02),核因子-κB(NF-κB)(1.20±0.05 vs.0.94±0.04)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)(0.72±0.02 vs.0.40±0.03)、活性含半胱氨酸的天冬氨酸蛋白水解酶-1、消皮素D N-末端片段(GSDMD-N)(0.71±0.03 vs.0.52±0.01)蛋白表达明显下降,NF-κB、NLRP3免疫组化染色评分明显下降,肿瘤坏死因子-α、白细胞介素-1β、白细胞介素-18含量明显下降,差异均有统计学意义(P<0.05)。结论TBI后早期应用RGFP966干预可以减轻神经细胞焦亡和炎症反应,发挥神经保护作用,其机制可能与抑制NF-κB/NLRP3/GSDMD通路激活有关。Objective To investigate the neuroprotective effect of histone deacetylase 3(HDAC3)specific inhibitor RGFP966 on traumatic brain injury(TBI)and its mechanism in rats.Methods Forty-eight SD rats were randomly divided into sham-operated group,TBI group,TBI+vehicle group and TBI+RGFP966 group(n=12).Rats in the later 3 groups accepted hydraulic impact brain injury to establish TBI models;and then,RGFP966(dissolved in 1%DMSO at a dose of 10 mg/kg)was injected intraperitoneally 30 min after modeling,twice a day for 3 d,in TBI+RGFP966 group;same amount of DMSO was injected into TBI+vehicle group at the same time.Three d after modeling,neurological function was tested by modified neurological severity score(mNSS),water content of brain tissues was detected by dry-wet weight method,proportion of injured neurons at the frontal cortical tissues on the affected side was detected by Nissl staining,expressions of HDAC3 and pyroptosis related proteins were detected by immunohistochemical staining and Western blotting,and serum content of inflammatory factors was detected by ELISA.Results Three d after modeling,compared with the TBI+vehicle group,the TBI+RGFP966 group had significantly decreased mNSS scores(9.83±0.75 vs.6.67±0.82),water content of the injured cerebral cortex(82.73%±0.36%vs.80.92%±0.66%),proportion of damaged neurons(75.60%±7.44%vs.55.87%±4.10%),and HDAC3 protein expression(0.67±0.09 vs.0.51±0.07),and significantly increased acetylated H3(Ace-H3)and acetylated H4(Ace-H4)protein expressions(0.81±0.02 vs.1.22±0.02;0.74±0.01 vs.1.07±0.02),and statistically decreased protein expressions of nuclear factor-κB(NF-κB,1.20±0.05 vs.0.94±0.04),NOD-like receptor thermal protein domain associated protein 3(NLRP3,0.72±0.02 vs.0.40±0.03),Caspase-1 containing cysteine(Caspase-1),dermatin D N-terminal fragment(GSDMD-N,0.71±0.03 vs.0.52±0.01),significantly decreased NF-κB and NLRP3 immunohistochemical staining scores,and significantly decreased serum contents of tumor necrosis factor-α,interleukin(IL)-1βand

关 键 词：创伤性脑损伤 RGFP966 细胞焦亡 炎症反应 

分 类 号：R651.15[医药卫生—外科学]