目录
缩略词 (1)
摘要 (2)
Abstract (4)
前言 (6)
材料与方法 (8)
1. 主要材料 (8)
2. 主要仪器及设备 (9)
3. 主要试剂配制 (10)
4. 实验分组 (13)
柏杨树
5. 实验方法 (13)
6. 统计学分析 (20)
实验结果 (21)
讨论 (28)
蛋糕的做法和配方
结论 (30)
参考文献 (31)鼻涕臭
综述 (34)
研究生期间取得的学术成果 (41)
致谢 (42)
缩略词
缩略词英文全称中文全称
PA Palmitic acid 棕榈酸
ROS Reactive oxygen species 活性氧
FUNDC1 FUN14 domain-containing protein 1 FUN14结构域蛋白1 ATP Adenosine triphosphate 腺苷三磷酸
Bax Bcl-2 assaciated X protein 前凋亡蛋白
DMSO Dimethyl sulfoxide 二甲基亚砜日本国庆
FBS Fetal bovine rum 胎牛血清
SDS Sodium dodecyl sulfonate 十二烷基磺酸钠
APS Ammonium persulfate 过硫酸铵
制片TEMED N,N,N',N'-tetramethylethylenediamine 四甲基乙二胺
DMEM Dulbecco's modified eagle's medium Dulbeco改良Eagle培养基OD Optical density 吸光度
CCK-8 Cell counting kit-8 细胞活性检测试剂盒TBST Tris buffered saline tween-20 TBST缓冲液
PBS Phosphate buffered saline 磷酸盐缓冲液
PMSF Phenylmethanesulfonyl fluoride 苯甲基磺酰氟
PVDF Polyvinylidene difluoride 聚偏乙烯氟化物
好书推荐排行榜BCL-2 B-cell leukemia/lymphoma 2 B细胞淋巴癌-2蛋白ECL Enhanced chemiluminescence 增强化学发光
AAV Adeno associated virus 腺相关病毒
FUNDC1对高脂诱导的人血管内皮细胞的保护作用研究
摘要
目的：本研究使用腺病毒过表达FUNDC1，再用棕榈酸处理EA.hy926人血管内皮细胞，建立高脂损伤内皮细胞模型，探讨FUNDC1对高脂损伤的EA.hy926人血管内皮细胞的作用和其可能机制。
商业策划书封面方法：（1）建立高脂诱导损伤的EA.hy926人血管内皮细胞模型：选择浓度分别为0.1、0.15、0.2、0.25、0.3、0.5、0.7 mmol/L棕榈酸溶液刺激EA.hy926人血管内皮细胞，刺激的时间为24 h。（2）FUNDC1对高脂损伤EA.hy926人血管内皮细胞的保护作用研究：体外培养EA.hy926人血管内皮细胞长至70%~80%，将细胞分为4组：对照组、高脂组（0.2 mmol/L棕榈酸）、高脂+空载组（转染空载
腺病毒预处理24 h+0.2 mmol/L棕榈酸）、高脂+FUNDC1组（转染FUNDC1腺病毒预处理24 h+0.2 mmol/L棕榈酸）进行CCK8实验检测各组细胞增殖活性；DCFH-DA活性氧检测试剂盒检测各组细胞活性氧含量变化情况；JC-1法测定各组细胞线粒体膜电位变化情况；ATP含量检测试剂盒检测各组细胞ATP的含量；蛋白印迹法检测各组细胞自噬（P62、LC3）和凋亡（Bcl-2、Bax）相关蛋白的表达情况。
结果：（1）与对照组相比，浓度为0.2 mmol/L的棕榈酸溶液刺激细胞，时间为24 h，细胞活力显著下降（P＜0.01）。（2）与对照组相比较，高脂组细胞活力显著降低(P＜0.01)；高脂+FUNDC1组存活率相比高脂组显著上升（P＜0.01），且高脂组和高脂+空载组两组无显著性差异（P＞0.05）。（3）与对照组相比，高脂组活性氧含量明显上升（P＜0.01）；高脂+FUNDC1组活性氧含量相比高脂组显著下降(P＜0.01)，且高脂组和高脂+空载组两组无显著性差异（P＞0.05）。（4）与对照组相比，高脂组线粒体膜电位去极化比例明显升高（P＜0.01）；高脂+FUNDC1组线粒体膜电位去极化比例相比高脂组显著下降（P＜0.01），且高脂组和高脂+空载组之间无显著差异（P＞0.05）。（5）与对照组相比，高脂组ATP含量明显下降（P＜0.01）；高脂+FUNDC1组ATP含量相比高脂组显著上升(P＜0.05)，且高脂组和高脂+空载组两组无显著性差异（P＞0.05）。（6）与对照组相比较，高脂组LC3、P62、Bax量显著上升（P＜0.05），Bcl-2量明显
投资决策降低（P＜0.05）；高脂+FUNDC1组LC3、P62、Bax量相比高脂组显著下降（P ＜0.05），Bcl-2量
明显升高（P＜0.05），且高脂组和高脂+空载组两组无显著性差异（P＞0.05）。
结论：（1）在高脂条件下，EA.hy926血管内皮细胞存活率下降、ROS含量升高、线粒体膜电位降低、ATP含量降低，过表达FUNDC1能够改善高脂对EA.hy926血管内皮细胞的损伤情况。（2）过表达FUNDC1可促进EA.hy926血管内皮细胞发生自噬，减少凋亡蛋白表达。
关键词：FUNDC1，棕榈酸，内皮细胞，高脂损伤，自噬
Protective effect of FUNDC1 on human vascular endothelial
cells damaged by lipotoxicity
Abstract
Objective: This study ud adenovirus to overexpress FUNDC1 and treat EA.hy926 human vascular endothelial cells with palmitic acid to establish a model of high-fat injury endothelial cells. To explore the effect of FUNDC1 on high-fat injury EA.hy926 human vascular endothelial cells and its role possible mechanism. Methods: (1)Establish EA.hy926 human vascular endothelial cell model with high-fat damage: lect concentrations of 0.1, 0.15, 0.2, 0.25, 0.3, 0.5, 0.7 mmol/L palmitic acid solution to stimulate EA.hy926 cells, palmitic acid stimulated the cells for 24 h.
(2)Protective effects of FUNDC1 on high-fat injury EA.hy926 cells: In vitro culture of EA.hy926 human vascular endothelial cells grew to about 70%-80%. There are four groups of cells: control group, high-fat Group (0.2 mmo/L palmitic acid), high-fat+empty group (transfection with empty adenovirus pretreatment for 24 h+0.2 mmol/L palmitic acid), high-fat+FUNDC1 group (transfection with FUNDC1 adenovirus pre-treatment for 24 h+0.2 mmol/L palmitic acid) performed CCK8 test to detect cell proliferation activity in each group; DCFH-DA reactive oxygen kit to detect changes in reactive oxygen content of cells in each group; Changes of mitochondrial membrane potential measured by JC-1 method; The kit was ud to detect the ATP content of each group of cells; Detection of P62, LC3, Bcl-2, Bax in each group by western blot.
Results: (1)Cells with a concentration of 0.2 mmol/L of palmitic acid stimulated the cells for 24 h, and the cell viability was significantly reduced compared with the control group(P<0.01). (2)The cell viability of the high-fat group and the high-fat+empty group decread significantly compared with the control group(P<0.01), between the high-fat group and the high-fat+empty group, there was no significant difference(P>0.05); The high-fat+FUNDC1 group survival rate was significantly higher than that of the high-fat group(P<0.01). (3)The content of active oxygen in the high-fat group and the high-fat+empty group incread significantly
