靶向AQP4的RNAi对大鼠脑出血周围细胞凋亡影响
发布时间：2018-11-13 08:51
【摘要】：研究背景、目的:脑出血是老年患者常见的一种疾病,具有极高的致死率及致残率,严重威胁人的生命。目前每年因脑出血死亡的患者据统计约占全部疾病死亡的20%左右。脑出血对脑细胞造成的损害目前尚无有效预防治疗措施,血肿的占位效应和其对周围脑组织的直接破坏是出血性脑损伤的两种主要机制,继发性损伤也是影响脑出血预后的重要因素之一。国内外的研究证实,出血后脑血肿的周边脑组织中存在不完全性缺血区,而不完全性缺血损伤则可能导致细胞凋亡,从而产生继发性脑组织损伤。细胞凋亡即参与了脑出血后的继发性损伤。细胞凋亡的过程包含了诱导启动、细胞内调控、细胞内实施、细胞的吞噬搬运四个阶段。其特点之一即是水的流失以及细胞体积的缩小,而水的流失主要以水分子穿过疏水的磷脂双分子层单纯扩散和依靠具体的水通道蛋白转运两种方式穿过细胞膜流到细胞外,由此可以看出,脑出血后血肿周围细胞凋亡可能与水通道蛋白有关。水通道蛋白(aquaporin,AQP)是一种细胞内运输水的分子通道,迄今为止在哺乳动物组织中已发现了13种水通道蛋白(AQP0~AQP12),其中水通道蛋白4(aquaporin4,AQP4)在哺乳动物脑组织中含量较丰富,参与了出血后脑水肿的形成和消退过程,在细胞凋亡过程中对水分子穿过细胞膜的运动起着非常重要的调节作用。据此本课题组提出能否通过干扰AQP4表达,在减轻脑水肿的同时减轻脑细胞的凋亡,从而保护脑血肿周边脑组织,减少继发性损害。目的:研究RNA干扰AQP-4表达对大鼠脑出血周围细胞凋亡影响。方法:30只SD大鼠分为脑出血模型组、空载质粒组、AQP-4 si RNA干预组组,每组10只。凝血酶Ⅶ注射诱发尾壳核脑的出血模型,造模后三天采用改良的Longa分级法进行神经功能评分,脑组织含水量测定,Tunel法检测脑组织凋亡情况,RT-PCR法检测血肿周围脑组织AQP-4表达,RT-PCR法检测血肿周围脑组织AQP-4表达,western-blot检测脑出血周围脑组织MMP-2,MMP-9,Caspase-3,Bcl-2表达。结果:AQP-4 si RNA干预组神经功能评分明显低于模型组和空载质粒组,比较有统计学意义(P0.05);模型组与空载质粒组大鼠脑组织出血明显,脑组织出血周围组织形成明显脑组织水肿,而AQP-4 si RNA干预组含水量在76.7%,统计学结果显示具有显著差异(P0.05);Tunel结果显示模型组与空载质粒组脑组织出血周围组织染色呈棕黄色区域较多。AQP-4 si RNA干预组染色较浅;模型组与空白质粒组大鼠脑组织出血周围组织AQP-4 RNA表达明显增加,表达明显高于AQP-4 si RNA干预组,比较有统计学意义(P0.05);AQP-4 si RNA干预组大鼠脑组织出血周围组织MMP-2/MMP-9表达明显降低,Caspase-3表达明显降低,Bcl-2表达增加,与模型组与空白质粒组比较有统计学意义(P0.05)。结论:RNA干扰AQP-4表达明显减轻大鼠脑出血大鼠神经功能损伤,抑制脑出血周围细胞凋亡,可能与减少AQP-4表达以及凋亡因子MMP-2,MMP-9,Caspase-3表达,增加Bcl-2表达有关。
[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，
本文编号：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，
本文编号：2328611
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