CAJ | 학술논문

背景研究表明,轻度修饰低密度脂蛋白与动脉粥样硬化的形成有关,除了具有蓄积低密度脂蛋白作用外还有很强的生物学活性,可在培养细胞及动物体内诱导巨噬细胞集落刺激因子等多种生物活性物质的表达.目的:采用差异显示-聚合酶链反应技术研究轻度修饰低密度脂蛋白诱导血管内皮细胞的基因表达差异,为进一步阐明轻度修饰低密度脂蛋白与动脉粥样硬化的关系奠定基础.设计:重复测量设计.单位:泰山医学院.材料:实验于2003-07/2004-07在泰山医学院基础研究所完成.人脐静脉内皮细胞培养基为M199,在37℃,50 mL/L的CO2条件下培养.细胞生长到融合状态时,培养基中加入轻度修饰低密度脂蛋白,至终浓度为400mg/L,诱导处理30 h.方法:用差异显示反转录-聚合酶链反应技术分析轻度修饰低密度脂蛋白诱导下人血管内皮细胞的基因表达差异,并用反向Northern分析证实差异显示基因片段.主要观察指标:①内皮细胞mRNA的差异显示分析.②差异片段的克隆、序列分析及同源性比较.③诱导与非诱导小鼠肝脏mRNA的反向Northern分析.结果:轻度修饰低密度脂蛋白诱导下人血管内皮细胞出现一些上调和下调的基因片段.上调基因有胸腺素β4、FGFRI原癌基因伴随蛋白、FK506结合蛋白、rTSβ蛋白和细胞间粘附分子1;下调的基因有Apobec-1结合蛋白1、细胞色素B561和ERP72.结论:用差异显示反转录-聚合酶链反应方法证实在体外轻度修饰低密度脂蛋白可诱导人脐静脉内皮细胞一些基因的表达水平变化,引起轻度修饰低密度脂蛋白血管内皮细胞发生病理变化,最终导致动脉粥样硬化斑块的形成.
배경연구표명,경도수식저밀도지단백여동맥죽양경화적형성유관,제료구유축적저밀도지단백작용외환유흔강적생물학활성,가재배양세포급동물체내유도거서세포집락자격인자등다충생물활성물질적표체.목적:채용차이현시-취합매련반응기술연구경도수식저밀도지단백유도혈관내피세포적기인표체차이,위진일보천명경도수식저밀도지단백여동맥죽양경화적관계전정기출.설계:중복측량설계.단위:태산의학원.재료:실험우2003-07/2004-07재태산의학원기출연구소완성.인제정맥내피세포배양기위M199,재37℃,50 mL/L적CO2조건하배양.세포생장도융합상태시,배양기중가입경도수식저밀도지단백,지종농도위400mg/L,유도처리30 h.방법:용차이현시반전록-취합매련반응기술분석경도수식저밀도지단백유도하인혈관내피세포적기인표체차이,병용반향Northern분석증실차이현시기인편단.주요관찰지표:①내피세포mRNA적차이현시분석.②차이편단적극륭、서렬분석급동원성비교.③유도여비유도소서간장mRNA적반향Northern분석.결과:경도수식저밀도지단백유도하인혈관내피세포출현일사상조화하조적기인편단.상조기인유흉선소β4、FGFRI원암기인반수단백、FK506결합단백、rTSβ단백화세포간점부분자1;하조적기인유Apobec-1결합단백1、세포색소B561화ERP72.결론:용차이현시반전록-취합매련반응방법증실재체외경도수식저밀도지단백가유도인제정맥내피세포일사기인적표체수평변화,인기경도수식저밀도지단백혈관내피세포발생병리변화,최종도치동맥죽양경화반괴적형성.
BACKGROUND: It is indicated in research that mildly modified low density lipoprotein (mm-LDL) is related to atherogenesis and it not only stores LDL and provides very strong biological activity, but also expresses many kinds of bioactive substances, like macrophage colony stimulating factor (MCSF) induced in cell culture and in animal body.OBJECTIVE: Differential display-PCR (DD-PCR) technique is used to study the genetic expressed difference in mm-LDL inducing vascular endothelial cells so as to lay the foundation for further explanation of the relationship between mm-LDL and arteriosclerosis.DESIGN: Repeated measurement was designed.SETTING: Taishan Medical College.MATERIALS: The experiment was performed in Basic Institute of Taishan Medical College from July 2003 to July 2004. Medium of human umbilical vein endothelial cells (HUVECs) was M199. Culture was done at 37 ℃, in 50 mL/L CO2. When cells grew to the fusion state, mm-LDL was added in the medium to the terminal concentration of 400 mg/L and then,induction was followed in 30 hours.METHODS: DD- reverse transcription (RT)-PCR technique was used to analyze genetic expression difference of human vascular endothelial cells induced with mm-LDL and reverse Northern analysis was performed to testify DD genetic fragments.mRNA in liver of mice.RESULTS: Human vascular endothelial cells induced with inm-LDL displayed some up and down-regulated genetic fragments. Up-regulated genes included thymosin 34, FGFRI protooncogene-chaperone protein, FK506 binding protein, rTSβ protein and intercellular adhesion molecule-1 (Ⅰ-CAM-1). Down-regulated genes included Apo bec-1 binding protein-l, cytochromeB561 and ERP72.CONCLUSION: DDRT-PCR testifies that mm-LDL induces changes of some genetic expression of human umbilicus vein endothelial cells in vitro and pathological changes of mm-LDL vascular endothelial cells, terminally results in atherogenesis.