CAJ | 학술논문

目的应用同位素标记相对和绝对定量(iTRAQ)技术对糖尿病小鼠视网膜蛋白组学进行研究并对细胞凋亡相关蛋白进行初步分析. 方法 8只雄性糖尿病小鼠为糖尿病组(DM组),8只同龄正常小鼠为对照组.饲以标准颗粒饲料并分笼喂养,自由饮食,观察10周.实验结束时,禁食12 h处死,迅速分离视网膜,苏木精-伊红(HE)染色制作光镜组织切片观察病理结构改变.应用iTRAQ技术筛选鉴定差异表达蛋白,对细胞凋亡相关蛋白进行初步分析. 结果光镜下对照组小鼠视网膜各层排列整齐,结构清晰,糖尿病组小鼠视网膜各层结构疏松,组织水肿和毛细血管扩张现象明显.蛋白质组学筛选鉴定差异蛋白共348个,其中细胞凋亡相关的差异蛋白共16个,包括N-myc下游调节基因1蛋白、黏蛋白4、水通道蛋白1、信号转导和转录激活因子5B等.其中在糖尿病组小鼠视网膜表达上调者8个,下调者8个.Ingenuity Pathway Analysis(IPA)软件显示部分蛋白间交互作用网络. 结论细胞凋亡参与了糖尿病视网膜病变的发生发展,筛选鉴定该过程相关差异蛋白可为进一步的研究奠定基础.
목적 응용동위소표기상대화절대정량(iTRAQ)기술대당뇨병소서시망막단백조학진행연구병대세포조망상관단백진행초보분석. 방법 8지웅성당뇨병소서위당뇨병조(DM조),8지동령정상소서위대조조.사이표준과립사료병분롱위양,자유음식,관찰10주.실험결속시,금식12 h처사,신속분리시망막,소목정-이홍(HE)염색제작광경조직절편관찰병리결구개변.응용iTRAQ기술사선감정차이표체단백,대세포조망상관단백진행초보분석. 결과 광경하대조조소서시망막각층배렬정제,결구청석,당뇨병조소서시망막각층결구소송,조직수종화모세혈관확장현상명현.단백질조학사선감정차이단백공348개,기중세포조망상관적차이단백공16개,포괄N-myc하유조절기인1단백、점단백4、수통도단백1、신호전도화전록격활인자5B등.기중재당뇨병조소서시망막표체상조자8개,하조자8개.Ingenuity Pathway Analysis(IPA)연건현시부분단백간교호작용망락. 결론 세포조망삼여료당뇨병시망막병변적발생발전,사선감정해과정상관차이단백가위진일보적연구전정기출.
Objective To analyze the protein expression changes of retina in diabetic mice using isobaric tags for relative and absolute quantitation (iTRAQ) approach and to study proteins of apoptosis.Methods 8 diabetic mice were chosen as the diabetic model group (DM group),8diabetic mice as the normal control group.The animals were housed in wire-bottomed cages and received normal pellet chow and tap water in a constant environment.After 10 weeks,all mice were killed,and their retina were dissected.After hematoxylin and eosin(H&E) staining,the sections of retina were examined using light microscopy.The changes of protein expression in retina were studied using iTRAQ approach.Expression of apoptosis associated proteins was analyzed using ingenuity pathway analysis (IPA).Results Compared with control group,mice retina in DM group developed looser structures,tissue edema and obvious telangiectasia under light microscopy.Using iTRAQ approach,a total of 348 differential proteins were identified.Among those proteins,16 proteins were related with apoptosis,including Ataxin-10,Protein NDRG1,mucin-4,Aquaporin-1 and annexin A4,etc.There were 8 apoptosis-related proteins in retina with up-regulation,and the other 8 proteins with down-regulation in the DM group.The relationship between these proteins were analyzed and charted by IPA.Conclusions Apoptosis may be involved in the development of diabetic retinopathy.The identification of the apoptosis-related proteins will be helpful for the further study.