糖尿病心肌病内质网应激相关的差异表达蛋白分析  

Proteomic analysis of differentially expressed proteins related to endoplasmic reticulum stress in the diabetic myocardium

作　　者：秦瑞婕 李保应[1] 栾思思 李小利[1] 于飞[1] 蔡茜[1] 程梅[1] 高海青[1] 

机构地区：[1]山东大学齐鲁医院老年病科山东省心血管蛋白质组学重点实验室,济南250012

出　　处：《中华老年医学杂志》2014年第10期1122-1126,共5页Chinese Journal of Geriatrics

基　　金：基金项目：国家科技重大专项”十二五”计划（2012ZX09303016-003）,国家自然科学基金（81100595）

摘　　要：目的 应用同位素标记的相对和绝对定量技术（iTRAQ）检测和探讨糖尿病心肌病内质网应激相关的差异表达蛋白质的变化. 方法 以雄性C57BLKS/J db/db小鼠为2型糖尿病模型组（8只）,db/m小鼠为正常对照组（8只）;每周测定体质量、微量血糖变化.于18周末留取血清测定空腹血糖、总胆固醇、三酰甘油、糖基化终末产物（AGEs）的含量;留取心肌组织行HE染色观察组织病理变化,透射电镜观察肌浆网超微结构变化,应用iTRAQ技术检测各组内质网应激相关的差异表达蛋白. 结果 模型组与对照组比较,第8～18周体质量显著增加（P＜0.01）;第18周空腹血糖、总胆固醇、三酰甘油及血清AGEs含量显著升高（P＜0.01）.模型组小鼠心肌组织HE染色可见心肌细胞肥大、肌纤维排列不规则、肌丝断裂、并伴有核损伤;超微结构观察显示模型组心肌组织肌小节失去正常结构,明暗带消失,肌浆网扩张、肿胀,肌纤维间水肿,其间可见炎性细胞浸润.模型组与正常对照组心肌组织应用质谱分析结合数据库检索,共鉴定了22个与内质网应激相关的差异表达蛋白,其中上调者6个,下调者16个. 结论 内质网应激相关差异表达蛋白含缬酪肽蛋白、B细胞受体相关蛋白31、溶血磷脂酰胆碱酰基转移酶3,在糖尿病心肌病中具有重要作用,并可能为研究或发现糖尿病心肌病防治的新药靶点提供理论基础.Objective To investigate the differentially expressed proteins related to endoplasmic reticulum stress in myocardial tissues of diabetic mice using isobaric tags for relative and absolute quantitation （iTRAQ）.Methods Male C57BLKS/J db/db mice were chosen as the diabetic model group （the DM group,n=8）,while male db/m mice served as the control group （the CC group,n=8）.The mice were weighed every week for a total of ten weeks.At the end of the experiments,all mice were sacrificed.Fasting blood glucose （FBG）,total cholesterol （TC）,triglycerides （TG）,and serum advanced glycation end products （AGEs） were measured.Hematoxylin-eosin （H＆amp;E） staining was used to examine pathological changes of the myocardial tissue and transmission electron microscopy was used to examine ultrastructural changes.In addition,poteomics of the heart tissue extracts by iTRAQ analysis was obtained from both groups.Results Compared with the CC group,the DM group showed greater body weight at week 8,10,12,14,16,18 （P〈0.01 for all） and higher serum FBG,TC,TG and AGEs levels at week 18 （P〈 0.01 for all）.Examination with H＆amp;E staining revealed myocardial hypertrophy,myofiber disarray,and myofibril fractures accompanied by damaged nuclei in the DM group.Moreover,ultramicroscopic abnormalities,including irregular sarcomere structure,disappearance of the light zones and dark bands and sarcoplasmic reticulum dilation,swelling and edema,interspersed with inflammatory cells,were also seen in the DM group.Twenty-two differentially expressed proteins in myocardial tissue were identified through proteomic profile analysis and database search.Of these proteins,six were upregulated and 16 were down-regulated in the db/db mice.Conclusions The proteins related to endoplasmic reticulum stress,such as valosin containing protein,B-cell receptor-associated protein 31 and lysophosphatidylcholine acyltransferase 3,play an important role in diabetic cardiomyopathy and may provide a fundamental basis for studying or

关 键 词：心肌病 糖尿病 2型 糖基化终产物 高级 内质网 应激 

分 类 号：R587.2[医药卫生—内分泌]