本文選題：顱腦損傷 + 冷誘導(dǎo)RNA結(jié)合蛋白�。� 參考：《山東醫(yī)藥》2016年42期

【摘要】：目的探討冷誘導(dǎo)RNA結(jié)合蛋白(CIRP)表達(dá)在顱腦損傷(TBI)大鼠亞低溫(MH)治療過程中的作用,并分析其機(jī)制。方法將大鼠隨機(jī)分為A、B、C組,分別一次性鞘內(nèi)注射0.1 m L生理鹽水、空白AD5-GFP、AD5-GFP-CIRP-siRNA;每組再分為4個(gè)亞組,即假手術(shù)組(Sham組)、TBI組、MH組、TBI+MH組。TBI、TBI+MH組采用液壓打擊裝置進(jìn)行TBI造模,Sham組及MH組不予打擊;TBI造模成功后,MH組、TBI+MH組行MH治療48 h。于MH開始治療后30 min及6、12、24、48、72 h,采用Western blot法檢測(cè)下丘腦部位CIRP、Ras、Raf、ERK-1/2、pERK-1/2蛋白;MH開始治療后96 h取大鼠腦皮層、海馬、下丘腦部位腦組織,采用TUNEL法檢測(cè)凋亡腦細(xì)胞、RTPCR法檢測(cè)CIRP mRNA。結(jié)果 A、B、C組中,TBI、TBI+MH組皮層、海馬、下丘腦部位神經(jīng)細(xì)胞凋亡指數(shù)均較Sham、MH組升高(P均0.05);A、B組中,TBI+MH組各部位神經(jīng)細(xì)胞凋亡指數(shù)較TBI組降低(P均0.05),且該組下丘腦部位較皮層、海馬區(qū)神經(jīng)細(xì)胞凋亡指數(shù)下降明顯(P均0.05);在C組中,TBI+MH組各部位神經(jīng)細(xì)胞凋亡指數(shù)較TBI組無差異(P均㧐0.05)。在A、B組中,MH、TBI+MH組各部位CIRP mRNA較Sham組升高(P均0.05),下丘腦部位較皮層、海馬區(qū)CIRP mRNA表達(dá)升高(P均0.05);而在C組中,TBI、MH、TBI+MH組各部位CIRP mRNA較Sham組均無差異(P均㧐0.05),各部位CIRP mRNA表達(dá)無差異(P均㧐0.05)。在A、B組中,TBI組CIRP蛋白自創(chuàng)傷后12、24 h較創(chuàng)傷后30 min升高(P均0.05),隨后開始下降,在48、72 h無差異(P均0.05);MH、TBI+MH組CIRP蛋白表達(dá)自MH開始后6 h升高(P均0.05),于48 h達(dá)高峰,72 h呈下降趨勢(shì);在C組中,各組CIRP蛋白表達(dá)均無差異(P均0.05)。TBI、MH、TBI+MH組Ras激活度(Ras/Raf值)均進(jìn)行性升高,于24 h達(dá)到峰值,隨后進(jìn)行性下降;MH、TBI+MH組可導(dǎo)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組中MH、TBI+MH組此作用消失。結(jié)論在MH治療TBI過程中,CIRP因低溫作用而在皮層、海馬、下丘腦部位過表達(dá),其中下丘腦部位更加明顯;CIRP作用機(jī)制可能通過直接調(diào)控ERK-1/2激活,使其激活度隨時(shí)間推移迅速降低,減少對(duì)應(yīng)部位腦組織細(xì)胞凋亡,從而發(fā)揮神經(jīng)保護(hù)作用。
[Abstract]:Objective to investigate the role of cold-induced RNA-binding protein (CIRP) expression in the treatment of mild hypothermia (MH) in rats with craniocerebral injury (TBI) and its mechanism. Methods the rats were randomly divided into two groups: group A, B, C, and intrathecal injection of 0.1 mL normal saline respectively, and the rats were divided into 4 subgroups, each group was treated with AD5-GFPN AD5-GFP-CIRP-siRNAs, and each group was divided into 4 subgroups. That is, sham-operated group (Sham group), TBI group (MH group), TBI group (TBI group), TBI group, TBI group. At 30 min after treatment and 61212 ~ (24) and 48 ~ 72 h after treatment, the rat cerebral cortex, hippocampus and hypothalamus brain tissues were detected by Western blot method with Western blot method. The apoptotic brain cells were detected by Tunel and RT PCR of apoptotic brain cells (RT PCR) by using Tunel method. The rat brain cortex, hippocampus and hypothalamus brain tissues were detected by Western blot method. The rat brain cortex, hippocampus and hypothalamus were detected by Western blot method using Western blot method. The rat brain cortex, hippocampus and hypothalamus brain tissues were detected by RT-PCR with Western blot method. Results the apoptotic index of neurons in the cortex, hippocampus and hypothalamus of TBI group was significantly higher than that in the Shambone MH group (P 0.05). The apoptotic index of neurons in TBI group was lower than that in TBI group (P 0.05), and the apoptotic index in hypothalamus was lower in TBI group than in TBI group (P 0.05), and the apoptotic index in hypothalamus was lower in TBI group than in TBI group (P 0.05). The apoptotic index of hippocampal neurons decreased significantly (all P 0.05), and there was no significant difference between TBI MH group and TBI group (P 0.05). The CIRP mRNA expression in the hypothalamus was higher than that in the cortex and hippocampus (P 0.05), but there was no significant difference in the CIRP mRNA expression between the TBI group and the Sham group (P 0.05), but there was no significant difference in the CIRP mRNA expression between the TBI group and the Sham group (P < 0.05), while the expression of CIRP mRNA in the hypothalamus was higher than that in the cortex and hippocampus (P 0.05), while in the TBI group there was no significant difference between the two groups (P < 0.05), and there was no significant difference in the expression of CIRP mRNA between the two groups (P < 0.05). The expression of CIRP protein in TBI group increased from 1224 hours after trauma to 30 min after trauma (P 0.05), and then decreased. There was no significant difference (P 0. 05) at 48 ~ 72 h. The expression of CIRP protein in TBI group increased 6 hours after the onset of MHBI (all P 0. 05), and decreased at 48 h and 72 h after trauma. In group C, there was no difference in the expression of CIRP protein in all groups (P 0.05). The Ras activation (Ras-Raf value) in TBI group increased progressively and reached its peak value at 24 h, and then decreased progressively, which resulted in the peak value of ERK-1 / 2 activation (p-ERK-1 / 2 / ERK-1 / 2). The activation degree of MHG TBI MH group was decreased rapidly, but the effect of MHX TBI MH group disappeared in C group. Conclusion CIRP is overexpressed in cortex, hippocampus and hypothalamus due to hypothermia during the treatment of TBI. The mechanism of CIRP in hypothalamus is more obvious, and the activation degree of CIRP decreases rapidly over time through direct regulation of ERK-1 / 2 activation. Reduce the apoptosis of brain cells in the corresponding areas, thus play a neuroprotective role.
【作者單位】： 天津中醫(yī)藥大學(xué)第二附屬醫(yī)院;天津市環(huán)湖醫(yī)院神經(jīng)外科研究所;天津中醫(yī)藥大學(xué)研究生院;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(81303091)
【分類號(hào)】：R651.15

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