摘要
目的探讨冷诱导RNA结合蛋白(CIRP)表达在颅脑损伤(TBI)大鼠亚低温(MH)治疗过程中的作用,并分析其机制。方法将大鼠随机分为A、B、C组,分别一次性鞘内注射0.1 m L生理盐水、空白AD5-GFP、AD5-GFP-CIRP-siRNA;每组再分为4个亚组,即假手术组(Sham组)、TBI组、MH组、TBI+MH组。TBI、TBI+MH组采用液压打击装置进行TBI造模,Sham组及MH组不予打击;TBI造模成功后,MH组、TBI+MH组行MH治疗48 h。于MH开始治疗后30 min及6、12、24、48、72 h,采用Western blot法检测下丘脑部位CIRP、Ras、Raf、ERK-1/2、pERK-1/2蛋白;MH开始治疗后96 h取大鼠脑皮层、海马、下丘脑部位脑组织,采用TUNEL法检测凋亡脑细胞、RTPCR法检测CIRP mRNA。结果 A、B、C组中,TBI、TBI+MH组皮层、海马、下丘脑部位神经细胞凋亡指数均较Sham、MH组升高(P均<0.05);A、B组中,TBI+MH组各部位神经细胞凋亡指数较TBI组降低(P均<0.05),且该组下丘脑部位较皮层、海马区神经细胞凋亡指数下降明显(P均<0.05);在C组中,TBI+MH组各部位神经细胞凋亡指数较TBI组无差异(P均﹥0.05)。在A、B组中,MH、TBI+MH组各部位CIRP mRNA较Sham组升高(P均<0.05),下丘脑部位较皮层、海马区CIRP mRNA表达升高(P均<0.05);而在C组中,TBI、MH、TBI+MH组各部位CIRP mRNA较Sham组均无差异(P均﹥0.05),各部位CIRP mRNA表达无差异(P均﹥0.05)。在A、B组中,TBI组CIRP蛋白自创伤后12、24 h较创伤后30 min升高(P均<0.05),随后开始下降,在48、72 h无差异(P均>0.05);MH、TBI+MH组CIRP蛋白表达自MH开始后6 h升高(P均<0.05),于48 h达高峰,72 h呈下降趋势;在C组中,各组CIRP蛋白表达均无差异(P均>0.05)。TBI、MH、TBI+MH组Ras激活度(Ras/Raf值)均进行性升高,于24 h达到峰值,随后进行性下降;MH、TBI+MH组可导致ERK-1/2激活度(p-ERK-1/2/ERK-1/2值)峰值前移,并迅速降低其激活度,而C组中MH、TBI+MH组此作用消失。结论在MH治疗TBI过程中,CIRP因低温作用而在皮层、海马
Objective To investigate the effects of cold-inducible RNA-binding protein （CIRP）expression in the mild hypothermia （MH）therapeutic process for rats with traumatic brain injury （TBI）and to analyze its mechanism.Methods The rats were randomly divided into three groups：groups A,B and C,which were respectively injected with 0.1 mL normal saline, blank AD5-GFP and AD5-GFP-CIRP-SiRNA.Then,each group was subdivided into four groups：sham group ,TBI group,MH group,and TBI ＋MH groups.TBI models were made in the TBI and TBI ＋MH groups.After successfully modeling,the rats in MH group and TBI ＋MH group received MH treatment for 48 h.After MH of 30 min,6,12,24,48 and 72 h,we detected CIRP,Ras,Raf,ERK-1 /2 and p-ERK-1 /2 protein in the hypothalamus.After 96 hours of MH treatment,we took the cerebral cortex,hippocampus,and hypothalamus brain tissues.The brain apoptosis was detected by TUNEL,and the expression of CIRP mRNA was detected by RT-PCR.Results In the groups A,B and C,the apoptotic indexes of nerve cells in the cortex,hippo-campus and hypothalamus of TBI and TBI ＋MH group were higher than those of Sham and MH groups （all P 〈0.05）.In the groups A and B,the apoptotic index of neurons in all parts was significantly lower than that of the TBI group （all P 〈0.05）,the apoptotic index of neurons in hypothalamus was significantly decreased as compared with that in the hippocampus and hippocam-pus parts （all P 〈0.05）.In the groups A and B,CIRP mRNA in the MH,TBI ＋MH groups was higher than that in Sham group （all P 〈0.05）,CIRP mRNA expression in hypothalamus was higher than that in cortex and hippocampus （all P 〈0.05）.There was no significant difference in CIRP mRNA expression between the TBI,MH,TBI ＋MH groups and the Sham group （all P 〉0.05）as well as CIRP mRNA expression in all parts （all P 〉0.05）.In groups A and B,the CIRP protein in TBI group in-creased at 12 and 24 h after TBI as compared with that at 30 min （all P 〈0.05）,and then decreased at 48 and
出处
《山东医药》
CAS
北大核心
2016年第42期9-13,共5页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81303091)
关键词
颅脑损伤
冷诱导RN
A结合蛋白
亚低温
细胞凋亡
细胞外调节蛋白激酶
craniocerebral trauma
cold-inducible RNA-binding protein
mild hypothermia
apoptosis
extracellular signal-regulated kinase
