CAJ | 학술논문

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.
목적 응용동위소표기적상대화절대정량기술(iTRAQ)검측화탐토당뇨병심기병내질망응격상관적차이표체단백질적변화. 방법 이웅성C57BLKS/J db/db소서위2형당뇨병모형조(8지),db/m소서위정상대조조(8지);매주측정체질량、미량혈당변화.우18주말류취혈청측정공복혈당、총담고순、삼선감유、당기화종말산물(AGEs)적함량;류취심기조직행HE염색관찰조직병리변화,투사전경관찰기장망초미결구변화,응용iTRAQ기술검측각조내질망응격상관적차이표체단백. 결과 모형조여대조조비교,제8～18주체질량현저증가(P＜0.01);제18주공복혈당、총담고순、삼선감유급혈청AGEs함량현저승고(P＜0.01).모형조소서심기조직HE염색가견심기세포비대、기섬유배렬불규칙、기사단렬、병반유핵손상;초미결구관찰현시모형조심기조직기소절실거정상결구,명암대소실,기장망확장、종창,기섬유간수종,기간가견염성세포침윤.모형조여정상대조조심기조직응용질보분석결합수거고검색,공감정료22개여내질망응격상관적차이표체단백,기중상조자6개,하조자16개. 결론 내질망응격상관차이표체단백함힐락태단백、B세포수체상관단백31、용혈린지선담감선기전이매3,재당뇨병심기병중구유중요작용,병가능위연구혹발현당뇨병심기병방치적신약파점제공이론기출.
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&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&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 finding new drug targets for the prevention of diabetic cardiomyopathy.