转染tdp-25对sh-sy5y细胞自噬蛋白表达及细胞活力 凋亡的影响-effect of tdp - 25 transfection on autophagy protein expression and apoptosis of sh - sy5y cells.docxVIP

万方数据 万方数据 摘要 凋亡的影响。 方法 分别转染 GFP-N1、GFP-TDP-43 和 GFP-TDP-25 三种质粒的神经母细胞 瘤细胞 SH-SY5Y 分为三组，24h 后应用免疫荧光法检测包涵体的存在及其分布； 应用 Western blot 实验检测蛋白 LC3、Beclin-1 的表达情况；将 SH-SY5Y 细胞转 染质粒 GFP-TDP-25 后分为两组，一组加入 3-MA 自噬抑制剂，另一组不做处理， MTT 比色法检测细胞活力；Annexin-V-FITC/PI 检测细胞凋亡情况。 结果 1.GFP-N1 组细胞核、胞质中均可见绿色荧光信号；GFP-TDP-43 组绿色荧 光信号主要分布于细胞核，未见明显的 TDP-43 向胞质中异位，且未见明显的包 涵体形成；GFP-TDP-25 组较强的绿色荧光信号呈颗粒状分布于细胞质，形成了 包涵体； 2.与 GFP-N1 组和 GFP-TDP-43 组相比，GFP-TDP-25 组细胞中蛋白 Beclin-1 表达上调，LC3-Ⅱ/LC3-Ⅰ的比值升高，差异具有统计学意义（P0.05）； 3.与未加入 3-MA 组相比，加入 3-MA 后，过表达 TDP-25 细胞的细胞活力 降低、细胞凋亡率增高，差异具有统计学意义（P0.05）。 结论 截短型片段 TDP-25 能够在细胞中形成包涵体，过表达 TDP-25 通过诱导细 胞自噬水平上调来减弱其对细胞损害。调控自噬水平对肌萎缩性侧索硬化症和 额颞叶变性患者病情进展可能起到一定延缓作用。 关键词 TDP-25 ； 自 噬 ； 肌 萎 缩 侧 索 硬 化 ； 额 颞 叶 变 性 II Abstra Abstract Expression of autophagy proteins and cell vitality, apoptosis of SH-SY5Y cells transfected with TDP-25 Postgraduate Yushuai Zhou Supervisor: Prof. Junfang Teng Neurology The First Affiliated Hospital of Zhengzhou University ZhengZhou 450052 Background Abstract Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disease .Patients experience signs and symptoms of progressive muscle atrophy and weakness, increased fatigue, which typically lead to respiratory failure and death .Median survival is 2 to 4 years from onset ; only 5–10% of patients survive beyond 10 years. frontotemporal lobar degeneration (FTLD) is the second most common type of dementia disorders in people under the age of 65 years ，it can manifest as a spectrum of clinical syndromes,ranging from behavioural impairment to language or motor dysfunction.Onset occurs most commonly between the ages of 45 and 65 years,The mean duration of illness is 8 years, ranging from 2 years to 20 years .At present, No effective treatment is available.The overlap in the clinical symptoms and pathological characteristics of the nervous system between ALS and FTLD indicates that they are I likely to be the same kind of neurodegenerative diseases, only selectively affect different excitable ne