TRPC通道在Aβ诱导的阿尔茨海默病（AD）小鼠的蛋白表达研究
发布时间：2019-07-04 15:22
【摘要】：阿尔茨海默病(AD)是一种发生在老年人中的神经退行性疾病,临床上主要以进行性认知功能减退为主要表现,其典型的病理特征为β淀粉样蛋白(Aβ)为核心的老年斑、神经纤维缠结以及神经元死亡。目前公认的Aβ沉积是AD形成的最终通路,而钙稳态失调能够导致神经元结构以及功能发生改变,甚至导致神经元的不可逆性损伤。近年来的研究发现经典瞬时受体电位(TRPC)通道是一类位于细胞膜上的非选择性钙离子通道,在大脑神经元中高度表达,并且研究表明TRPC通道在神经系统中主要参与增殖、分化、凋亡、退行性变和突触可塑性等方面的功能。但是,在Aβ诱导的AD小鼠模型中,TRPC通道蛋白的表达情况以及功能机制目前还尚不清楚。目的探讨经典瞬时受体电位(TRPC)通道蛋白在Aβ1-42诱导的阿尔茨海默病(AD)小鼠海马区中的表达情况。方法1.雄性5周龄ICR小鼠36只,适应性喂养5天后,将其随机分为AD组和对照组,每组18只。2.应用Aβ1-42侧脑室注射制作AD小鼠模型,侧脑室注射第十天后进行Morris水迷宫定位航行和空间探索实验测定小鼠学习记忆能力,行为学检测后处死动物,采用逆转录聚合酶链反应(RT-PCR)方法检测海马区TRPC1～TRPC7mRNA的表达情况,运用免疫荧光和Western blot等实验技术检测小鼠海马区的TRPC4通道蛋白的表达。结果1.Morris水迷宫行为学检测结果显示,在定位航行实验中,从第三天开始,与对照组相比,AD组小鼠水迷宫测试的逃避潜伏期延长(P0.05),寻找平台游泳路程较长(P0.05),在空间探索实验中,与对照组相比,AD组在目标象限停留时间明显缩短(P0.01),穿越平台次数明显减少(P0.01)。2.RT PCR检测结果显示,在对照组及AD组中,TRPC1～TRPC7通道mRNA均有不同程度的表达,与对照组相比,AD组TRPC4mRNA表达明显增高(P0.01)。3.免疫荧光结果显示TRPC4通道蛋白广泛表达在海马区的神经细胞膜上,且与对照组相比,AD组TRPC4荧光强度增强。4.Western blot结果显示,与对照组相比,AD组TRPC4通道蛋白表达增高(P0.05)。结论侧脑室注射Aβ1-42寡聚体后,小鼠学习记忆功能减退,海马区TRPC4通道mRNA及蛋白表达含量均增加,表明TRPC通道参与到因Aβ沉积而导致的AD发病机制中。
[Abstract]:Alzheimer's disease (AD) is a neurodegenerative disease in the elderly, which is mainly characterized by progressive cognitive impairment. The typical pathological features of Alzheimer's disease are Alzheimer's disease (A 尾), neurofibrillar tangles and neuronal death. At present, A 尾 deposition is recognized as the final pathway of AD formation, and calcium homeostasis disorder can lead to changes in the structure and function of neurons, and even lead to irreversible damage of neurons. In recent years, it has been found that the classical transient receptor potential (TRPC) channel is a kind of non-selective calcium channel located on the cell membrane, which is highly expressed in brain neurons, and studies have shown that TRPC channel is mainly involved in proliferation, differentiation, apoptosis, degeneration and synaptic plasticity in the nervous system. However, the expression and functional mechanism of TRPC channel protein in AD mice induced by A 尾 are not clear at present. Objective to investigate the expression of classical transient receptor potential (TRPC) channel protein in hippocampus of (AD) mice with Alzheimer's disease induced by A 尾 1-42. Method 1. Thirty-six male 5-week-old ICR mice were randomly divided into AD group (n = 18) and control group (n = 18) after 5 days of adaptive feeding. The AD mouse model was established by intraventricular injection of A 尾 1-42. The learning and memory ability of the mice was measured by Morris water maze localization navigation and space exploration test 10 days after lateral ventricle injection. The animals were killed after behavioral examination. The expression of TRPC1~TRPC7mRNA in hippocampus was detected by reverse transcription polymerase chain reaction (RT-PCR), and the expression of TRPC4 channel protein in hippocampus was detected by immunofluorescence and Western blot. Results the results of 1.Morris water maze behavior test showed that from the third day, compared with the control group, the escape latency of the water maze test in the AD group was prolonged (P 0.05), and the swimming distance was longer (P 0.05). In the space exploration experiment, compared with the control group, the residence time in the target quadrant of the AD group was significantly shortened (P 0.01). The number of crossing the platform was significantly decreased (P 0.01). The results of 2.RT PCR showed that TRPC1~TRPC7 channel mRNA was expressed in different degrees in the control group and AD group, and the expression of TRPC4mRNA in the AD group was significantly higher than that in the control group (P 0.01). The results of immunofluorescence showed that TRPC4 channel protein was widely expressed in hippocampal nerve cell membrane, and the fluorescence intensity of TRPC4 in AD group was higher than that in control group. 4. Western blot results showed that the expression of TRPC4 channel protein in AD group was higher than that in control group (P 0.05). Conclusion after intracerebroventricular injection of A 尾 1 鈮，
本文编号：2510014
[Abstract]:Alzheimer's disease (AD) is a neurodegenerative disease in the elderly, which is mainly characterized by progressive cognitive impairment. The typical pathological features of Alzheimer's disease are Alzheimer's disease (A 尾), neurofibrillar tangles and neuronal death. At present, A 尾 deposition is recognized as the final pathway of AD formation, and calcium homeostasis disorder can lead to changes in the structure and function of neurons, and even lead to irreversible damage of neurons. In recent years, it has been found that the classical transient receptor potential (TRPC) channel is a kind of non-selective calcium channel located on the cell membrane, which is highly expressed in brain neurons, and studies have shown that TRPC channel is mainly involved in proliferation, differentiation, apoptosis, degeneration and synaptic plasticity in the nervous system. However, the expression and functional mechanism of TRPC channel protein in AD mice induced by A 尾 are not clear at present. Objective to investigate the expression of classical transient receptor potential (TRPC) channel protein in hippocampus of (AD) mice with Alzheimer's disease induced by A 尾 1-42. Method 1. Thirty-six male 5-week-old ICR mice were randomly divided into AD group (n = 18) and control group (n = 18) after 5 days of adaptive feeding. The AD mouse model was established by intraventricular injection of A 尾 1-42. The learning and memory ability of the mice was measured by Morris water maze localization navigation and space exploration test 10 days after lateral ventricle injection. The animals were killed after behavioral examination. The expression of TRPC1~TRPC7mRNA in hippocampus was detected by reverse transcription polymerase chain reaction (RT-PCR), and the expression of TRPC4 channel protein in hippocampus was detected by immunofluorescence and Western blot. Results the results of 1.Morris water maze behavior test showed that from the third day, compared with the control group, the escape latency of the water maze test in the AD group was prolonged (P 0.05), and the swimming distance was longer (P 0.05). In the space exploration experiment, compared with the control group, the residence time in the target quadrant of the AD group was significantly shortened (P 0.01). The number of crossing the platform was significantly decreased (P 0.01). The results of 2.RT PCR showed that TRPC1~TRPC7 channel mRNA was expressed in different degrees in the control group and AD group, and the expression of TRPC4mRNA in the AD group was significantly higher than that in the control group (P 0.01). The results of immunofluorescence showed that TRPC4 channel protein was widely expressed in hippocampal nerve cell membrane, and the fluorescence intensity of TRPC4 in AD group was higher than that in control group. 4. Western blot results showed that the expression of TRPC4 channel protein in AD group was higher than that in control group (P 0.05). Conclusion after intracerebroventricular injection of A 尾 1 鈮，
本文编号：2510014
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