靶向AQP4的RNAi對大鼠腦出血周圍細(xì)胞凋亡影響
發(fā)布時(shí)間：2018-11-13 08:51
【摘要】：研究背景、目的:腦出血是老年患者常見的一種疾病,具有極高的致死率及致殘率,嚴(yán)重威脅人的生命。目前每年因腦出血死亡的患者據(jù)統(tǒng)計(jì)約占全部疾病死亡的20%左右。腦出血對腦細(xì)胞造成的損害目前尚無有效預(yù)防治療措施,血腫的占位效應(yīng)和其對周圍腦組織的直接破壞是出血性腦損傷的兩種主要機(jī)制,繼發(fā)性損傷也是影響腦出血預(yù)后的重要因素之一。國內(nèi)外的研究證實(shí),出血后腦血腫的周邊腦組織中存在不完全性缺血區(qū),而不完全性缺血損傷則可能導(dǎo)致細(xì)胞凋亡,從而產(chǎn)生繼發(fā)性腦組織損傷。細(xì)胞凋亡即參與了腦出血后的繼發(fā)性損傷。細(xì)胞凋亡的過程包含了誘導(dǎo)啟動、細(xì)胞內(nèi)調(diào)控、細(xì)胞內(nèi)實(shí)施、細(xì)胞的吞噬搬運(yùn)四個階段。其特點(diǎn)之一即是水的流失以及細(xì)胞體積的縮小,而水的流失主要以水分子穿過疏水的磷脂雙分子層單純擴(kuò)散和依靠具體的水通道蛋白轉(zhuǎn)運(yùn)兩種方式穿過細(xì)胞膜流到細(xì)胞外,由此可以看出,腦出血后血腫周圍細(xì)胞凋亡可能與水通道蛋白有關(guān)。水通道蛋白(aquaporin,AQP)是一種細(xì)胞內(nèi)運(yùn)輸水的分子通道,迄今為止在哺乳動物組織中已發(fā)現(xiàn)了13種水通道蛋白(AQP0~AQP12),其中水通道蛋白4(aquaporin4,AQP4)在哺乳動物腦組織中含量較豐富,參與了出血后腦水腫的形成和消退過程,在細(xì)胞凋亡過程中對水分子穿過細(xì)胞膜的運(yùn)動起著非常重要的調(diào)節(jié)作用。據(jù)此本課題組提出能否通過干擾AQP4表達(dá),在減輕腦水腫的同時(shí)減輕腦細(xì)胞的凋亡,從而保護(hù)腦血腫周邊腦組織,減少繼發(fā)性損害。目的:研究RNA干擾AQP-4表達(dá)對大鼠腦出血周圍細(xì)胞凋亡影響。方法:30只SD大鼠分為腦出血模型組、空載質(zhì)粒組、AQP-4 si RNA干預(yù)組組,每組10只。凝血酶Ⅶ注射誘發(fā)尾殼核腦的出血模型,造模后三天采用改良的Longa分級法進(jìn)行神經(jīng)功能評分,腦組織含水量測定,Tunel法檢測腦組織凋亡情況,RT-PCR法檢測血腫周圍腦組織AQP-4表達(dá),RT-PCR法檢測血腫周圍腦組織AQP-4表達(dá),western-blot檢測腦出血周圍腦組織MMP-2,MMP-9,Caspase-3,Bcl-2表達(dá)。結(jié)果:AQP-4 si RNA干預(yù)組神經(jīng)功能評分明顯低于模型組和空載質(zhì)粒組,比較有統(tǒng)計(jì)學(xué)意義(P0.05);模型組與空載質(zhì)粒組大鼠腦組織出血明顯,腦組織出血周圍組織形成明顯腦組織水腫,而AQP-4 si RNA干預(yù)組含水量在76.7%,統(tǒng)計(jì)學(xué)結(jié)果顯示具有顯著差異(P0.05);Tunel結(jié)果顯示模型組與空載質(zhì)粒組腦組織出血周圍組織染色呈棕黃色區(qū)域較多。AQP-4 si RNA干預(yù)組染色較淺;模型組與空白質(zhì)粒組大鼠腦組織出血周圍組織AQP-4 RNA表達(dá)明顯增加,表達(dá)明顯高于AQP-4 si RNA干預(yù)組,比較有統(tǒng)計(jì)學(xué)意義(P0.05);AQP-4 si RNA干預(yù)組大鼠腦組織出血周圍組織MMP-2/MMP-9表達(dá)明顯降低,Caspase-3表達(dá)明顯降低,Bcl-2表達(dá)增加,與模型組與空白質(zhì)粒組比較有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:RNA干擾AQP-4表達(dá)明顯減輕大鼠腦出血大鼠神經(jīng)功能損傷,抑制腦出血周圍細(xì)胞凋亡,可能與減少AQP-4表達(dá)以及凋亡因子MMP-2,MMP-9,Caspase-3表達(dá),增加Bcl-2表達(dá)有關(guān)。
[Abstract]:Background of the study: The purpose of this study is that the cerebral hemorrhage is a common disease in the elderly, with extremely high morbidity and disability, which is a serious threat to human life. The number of patients with cerebral hemorrhage is estimated to be about 20% of the total death rate at present. The damage of the cerebral hemorrhage to the brain cells is not effective in the treatment of cerebral hemorrhage. The place-occupying effect of the hematoma and its direct damage to the surrounding brain tissue are two main mechanisms of the hemorrhagic brain injury, and the secondary injury is one of the important factors that affect the prognosis of the cerebral hemorrhage. The studies at home and abroad confirm that there is an incomplete ischemic area in the peripheral brain tissue of the hematoma after the hemorrhage, and the incomplete ischemic injury can lead to the apoptosis of the cells, thus causing the secondary brain tissue injury. Apoptosis is the secondary injury after cerebral hemorrhage. The process of cell apoptosis includes four stages of induction initiation, intracellular regulation, intracellular implementation, and phagocytosis of the cells. one of the characteristics is the loss of water and the reduction of the volume of the cells, and the loss of water is mainly caused by the simple diffusion of water molecules through the hydrophobic phospholipid double-molecular layer and the flow of the cell membrane through the cell membrane through the specific water channel protein transport, The apoptosis of the perihematoma after intracerebral hemorrhage may be related to the water channel protein. aquaporin (AQP) is a molecular channel for intracellular transport of water, 13 water channel proteins (AQP0 to AQP12) have been found in mammalian tissue so far, with aquaporin 4 (aquaporin 4, AQP4) rich in mammalian brain tissue, It is involved in the formation and regression of brain edema after hemorrhage, and plays a very important role in the movement of water molecules through the cell membrane during the process of cell apoptosis. Therefore, the research group can reduce the brain edema and reduce the apoptosis of brain cells by interfering with AQP4 expression, so as to protect the brain tissue of the brain and reduce the secondary damage. Objective: To study the effect of RNA interference (AQP-4) on the apoptosis of rats with cerebral hemorrhage. Methods: 30 SD rats were divided into two groups: cerebral hemorrhage model group, no-load plasmid group and AQP-4si RNA interference group. The hemorrhage model of the brain of the caudate shell was induced by the injection of thrombin 鈪，
本文編號：2328611
[Abstract]:Background of the study: The purpose of this study is that the cerebral hemorrhage is a common disease in the elderly, with extremely high morbidity and disability, which is a serious threat to human life. The number of patients with cerebral hemorrhage is estimated to be about 20% of the total death rate at present. The damage of the cerebral hemorrhage to the brain cells is not effective in the treatment of cerebral hemorrhage. The place-occupying effect of the hematoma and its direct damage to the surrounding brain tissue are two main mechanisms of the hemorrhagic brain injury, and the secondary injury is one of the important factors that affect the prognosis of the cerebral hemorrhage. The studies at home and abroad confirm that there is an incomplete ischemic area in the peripheral brain tissue of the hematoma after the hemorrhage, and the incomplete ischemic injury can lead to the apoptosis of the cells, thus causing the secondary brain tissue injury. Apoptosis is the secondary injury after cerebral hemorrhage. The process of cell apoptosis includes four stages of induction initiation, intracellular regulation, intracellular implementation, and phagocytosis of the cells. one of the characteristics is the loss of water and the reduction of the volume of the cells, and the loss of water is mainly caused by the simple diffusion of water molecules through the hydrophobic phospholipid double-molecular layer and the flow of the cell membrane through the cell membrane through the specific water channel protein transport, The apoptosis of the perihematoma after intracerebral hemorrhage may be related to the water channel protein. aquaporin (AQP) is a molecular channel for intracellular transport of water, 13 water channel proteins (AQP0 to AQP12) have been found in mammalian tissue so far, with aquaporin 4 (aquaporin 4, AQP4) rich in mammalian brain tissue, It is involved in the formation and regression of brain edema after hemorrhage, and plays a very important role in the movement of water molecules through the cell membrane during the process of cell apoptosis. Therefore, the research group can reduce the brain edema and reduce the apoptosis of brain cells by interfering with AQP4 expression, so as to protect the brain tissue of the brain and reduce the secondary damage. Objective: To study the effect of RNA interference (AQP-4) on the apoptosis of rats with cerebral hemorrhage. Methods: 30 SD rats were divided into two groups: cerebral hemorrhage model group, no-load plasmid group and AQP-4si RNA interference group. The hemorrhage model of the brain of the caudate shell was induced by the injection of thrombin 鈪，
本文編號：2328611
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