中华围产医学杂志
中華圍產醫學雜誌
중화위산의학잡지
CHINESE JOURNAL OF PERINATAL MEDICINE
2012年
3期
158-162
,共5页
胎儿生长迟缓%脂肪组织%PPARγ%膜蛋白质类%磷酸烯醇丙酮酸羧激酶(GTP)%细胞内信号肽和蛋白质类
胎兒生長遲緩%脂肪組織%PPARγ%膜蛋白質類%燐痠烯醇丙酮痠羧激酶(GTP)%細胞內信號肽和蛋白質類
태인생장지완%지방조직%PPARγ%막단백질류%린산희순병동산최격매(GTP)%세포내신호태화단백질류
Fetal growth retardation%Adipose tissue%PPAR gamma%Membrane proteins%Phosphoenolpyruvate carboxykinase (GTP)%Intracellular signaling peptides and proteins
目的 研究胎儿生长受限(fetal growth restriction,FGR)子鼠内脏脂肪组织中转录因子过氧化物酶体增殖物激活受体γ2(peroxisome proliferator activated receptor gamma 2,PPARγ2)及其靶基因——脂肪分化相关蛋白(adipose differentiation-related protein,ADFP)、磷酸烯醇式丙酮酸羧激酶1(phosphoenolpyruvate carboxykinase 1,PCK1)的表达变化,探讨其在成年肥胖和代谢综合征发生中的作用机制. 方法 采用孕期低蛋白饮食法建立大鼠FGR模型.以雄性子鼠为研究对象,采用实时荧光定量逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,PCR)和蛋白印迹实验检测3周龄和8周龄子鼠肾周脂肪组织中的PPARγ2基因及其靶基因ADFP、PCK1的mRNA和蛋白表达,与同龄正常对照组子鼠进行比较.数据采用独立样本t检验及Spearman直线相关分析. 结果 3周龄和8周龄FGR组子鼠肾周脂肪组织中PPARγ2 mRNA的转录水平明显高于同龄对照组子鼠(3周龄:2.43±0.38与1.03±0.11,t=2.74,P＜0.05；8周龄:2.17±0.34与1.07±0.14,t=2.49,P＜0.05),蛋白含量也明显增高(3周龄:1.07±0.17与0.41±0.06,t=3.01,P＜0.05;8周龄:1.29±0.20与0.68±0.09,t=2.62,P＜0.05),分别是对照组的(261±31)％和(190±24)％.3周龄和8周龄FGR组子鼠肾周脂肪组织中PPARγ2调控的靶基因ADFP mRNA(3周龄:1.87±0.29与1.02±0.12,t=2.20；8周龄:1.64±0.31与1.06±0.14,t=3.47)和PCK1mRNA含量(3周龄:2.03±0.26与0.98±0.08,t=2.97；8周龄:1.79±0.28与1.03±0.11,t=3.24)和蛋白表达水平(ADFP蛋白表达3周龄:0.43±0.06与0.14±0.03,t=3.01；8周龄:0.71±0.09与0.38±0.05,t=3.06；PCK1蛋白表达3周龄:0.82±0.10与0.39±0.05,t=2.65；8周龄:0.85±0.11与0.67±0.07,t=2.86)也均高于对照组(P均＜0.05),且2个靶基因的表达水平与PPARγ2表达均成正相关(mRNA表达水平:r=0.907和0.826,P均＜0.01；蛋白表达水平:r=0.763和0.805,P均＜0.05).结论 宫内发育的“程序化”影响使FGR子代内脏脂肪组织中PPARγ2表达增加,进而引起靶基因ADFP和PCK1的表达增加,引起脂肪细胞积聚,这可能是成年肥胖和代谢综合征发生的机制之一.
目的 研究胎兒生長受限(fetal growth restriction,FGR)子鼠內髒脂肪組織中轉錄因子過氧化物酶體增殖物激活受體γ2(peroxisome proliferator activated receptor gamma 2,PPARγ2)及其靶基因——脂肪分化相關蛋白(adipose differentiation-related protein,ADFP)、燐痠烯醇式丙酮痠羧激酶1(phosphoenolpyruvate carboxykinase 1,PCK1)的錶達變化,探討其在成年肥胖和代謝綜閤徵髮生中的作用機製. 方法 採用孕期低蛋白飲食法建立大鼠FGR模型.以雄性子鼠為研究對象,採用實時熒光定量逆轉錄-聚閤酶鏈反應(reverse transcription-polymerase chain reaction,PCR)和蛋白印跡實驗檢測3週齡和8週齡子鼠腎週脂肪組織中的PPARγ2基因及其靶基因ADFP、PCK1的mRNA和蛋白錶達,與同齡正常對照組子鼠進行比較.數據採用獨立樣本t檢驗及Spearman直線相關分析. 結果 3週齡和8週齡FGR組子鼠腎週脂肪組織中PPARγ2 mRNA的轉錄水平明顯高于同齡對照組子鼠(3週齡:2.43±0.38與1.03±0.11,t=2.74,P＜0.05；8週齡:2.17±0.34與1.07±0.14,t=2.49,P＜0.05),蛋白含量也明顯增高(3週齡:1.07±0.17與0.41±0.06,t=3.01,P＜0.05;8週齡:1.29±0.20與0.68±0.09,t=2.62,P＜0.05),分彆是對照組的(261±31)％和(190±24)％.3週齡和8週齡FGR組子鼠腎週脂肪組織中PPARγ2調控的靶基因ADFP mRNA(3週齡:1.87±0.29與1.02±0.12,t=2.20；8週齡:1.64±0.31與1.06±0.14,t=3.47)和PCK1mRNA含量(3週齡:2.03±0.26與0.98±0.08,t=2.97；8週齡:1.79±0.28與1.03±0.11,t=3.24)和蛋白錶達水平(ADFP蛋白錶達3週齡:0.43±0.06與0.14±0.03,t=3.01；8週齡:0.71±0.09與0.38±0.05,t=3.06；PCK1蛋白錶達3週齡:0.82±0.10與0.39±0.05,t=2.65；8週齡:0.85±0.11與0.67±0.07,t=2.86)也均高于對照組(P均＜0.05),且2箇靶基因的錶達水平與PPARγ2錶達均成正相關(mRNA錶達水平:r=0.907和0.826,P均＜0.01；蛋白錶達水平:r=0.763和0.805,P均＜0.05).結論 宮內髮育的“程序化”影響使FGR子代內髒脂肪組織中PPARγ2錶達增加,進而引起靶基因ADFP和PCK1的錶達增加,引起脂肪細胞積聚,這可能是成年肥胖和代謝綜閤徵髮生的機製之一.
목적 연구태인생장수한(fetal growth restriction,FGR)자서내장지방조직중전록인자과양화물매체증식물격활수체γ2(peroxisome proliferator activated receptor gamma 2,PPARγ2)급기파기인——지방분화상관단백(adipose differentiation-related protein,ADFP)、린산희순식병동산최격매1(phosphoenolpyruvate carboxykinase 1,PCK1)적표체변화,탐토기재성년비반화대사종합정발생중적작용궤제. 방법 채용잉기저단백음식법건립대서FGR모형.이웅성자서위연구대상,채용실시형광정량역전록-취합매련반응(reverse transcription-polymerase chain reaction,PCR)화단백인적실험검측3주령화8주령자서신주지방조직중적PPARγ2기인급기파기인ADFP、PCK1적mRNA화단백표체,여동령정상대조조자서진행비교.수거채용독립양본t검험급Spearman직선상관분석. 결과 3주령화8주령FGR조자서신주지방조직중PPARγ2 mRNA적전록수평명현고우동령대조조자서(3주령:2.43±0.38여1.03±0.11,t=2.74,P＜0.05；8주령:2.17±0.34여1.07±0.14,t=2.49,P＜0.05),단백함량야명현증고(3주령:1.07±0.17여0.41±0.06,t=3.01,P＜0.05;8주령:1.29±0.20여0.68±0.09,t=2.62,P＜0.05),분별시대조조적(261±31)％화(190±24)％.3주령화8주령FGR조자서신주지방조직중PPARγ2조공적파기인ADFP mRNA(3주령:1.87±0.29여1.02±0.12,t=2.20；8주령:1.64±0.31여1.06±0.14,t=3.47)화PCK1mRNA함량(3주령:2.03±0.26여0.98±0.08,t=2.97；8주령:1.79±0.28여1.03±0.11,t=3.24)화단백표체수평(ADFP단백표체3주령:0.43±0.06여0.14±0.03,t=3.01；8주령:0.71±0.09여0.38±0.05,t=3.06；PCK1단백표체3주령:0.82±0.10여0.39±0.05,t=2.65；8주령:0.85±0.11여0.67±0.07,t=2.86)야균고우대조조(P균＜0.05),차2개파기인적표체수평여PPARγ2표체균성정상관(mRNA표체수평:r=0.907화0.826,P균＜0.01；단백표체수평:r=0.763화0.805,P균＜0.05).결론 궁내발육적“정서화”영향사FGR자대내장지방조직중PPARγ2표체증가,진이인기파기인ADFP화PCK1적표체증가,인기지방세포적취,저가능시성년비반화대사종합정발생적궤제지일.
Objective To investigate the expression of transcription factor peroxisome proliferator activated receptor gamma 2 (PPARγ2) and its target genes--adipose differentiation-related protein (ADFP) and phosphoenolpyruvate carboxykinase1 (PCK1) in visceral fat of growth restricted rat offspring,in order to explore their effects on the incidence of obesity and metabolism syndrome.Methods Fetal growth restriction (FGR) rat model was established by maternal low-protein diet and only male offsprings were studied.The mRNA expressions of PPARγ2,ADFP and PCK1 in perirenal fat tissues were measured at 3-week (W3) and 8-week (W8)-old rats by real time quantitative reverse transcription-polymerase chain reaction (real-time-RT-PCR). PPARγ2,ADFP and PCK1 protein levels in the perirenal fat tissues were determined by Western blot.Statistical analysis was performed with t-test and Spearman linear correlation analysis. Results The PPARγ2 mRNA level in perirenal fat tissue of FGR rats at W3 and W8 were significantly elevated than that of the controls (W3:2.43±0.38 vs 1.03±0.11,t=2.74,P＜0.05; W8:2.17±0.34 vs 1.07±0.14,t=2.49,P＜0.05),and PPARγ2 protein content was also increased (W3:1.07±0.17 vs 0.41±0.06,t=3.01,P＜0.05;W8:1.29±0.20 vs 0.68±0.09,t=2.62,P＜0.05),which equal to (261 ±31) ％ and (190±24) ％ of control,respectively.Increased mRNA and protein levels of the target genes ADFP and PCK1 in perirenal fat tissue were also found in the FGR rats at W3 and W8 compared with the same aged control rats (ADFP mRNA at W3:1.87±0.29 vs 1.02±0.12,t=2.20;ADFP mRNA at W8:1.64±0.31 vs 1.06±0.14,t=3.47; PCK1 mRNA at W3:2.03±0.26 vs 0.98 ± 0.08,t =2.97 ; PCK1 mRNA at W8:1.79 ± 0.28 vs 1.03 ± 0.11,t =3.24 ; ADFP protein at W3:0.43 ± 0.06 vs 0.14 ±0.03,t=3.01 ; ADFP protein at W8:0.71 ± 0.09 vs 0.38 ± 0.05,t =3.06 ; PCK1 protein at W3:0.82±0.10 vs 0.39±0.05,t=2.65;PCK1 protein at W8:0.85±0.11 vs 0.67±0.07,t=2.86; all P＜0.05).The protein and mRNA expressions of ADFP and PCK1 were positively correlated with expression the levels of PPARγ2 (mRNA:r=0.907 and 0.826,both P＜0.01;protein:r=0.763 and 0.805,both P＜0.05). Conclusions "Intrauterine programming" increases the expression of PPARγ2 in perirenal adipose tissue of FGR individuals,which might contribute to the increased expressions of its target genes ADFP and PCK1,thereby causes the accumulation of adipose cells and subsequently promotes obesityand metabolism syndrome in adulthood.
