We investigated whether maternal over-nutrition during pregnancy and lactation affects the offspring’s lipid metabolism at weaning by assessing liver lipid metabolic gene expressions and analysing its mechanisms on the development of metabolic abnormalities. Female Sprague–Dawley rats were fed with standard chow diet (CON) or high-fat diet (HFD) for 8 weeks, and then continued feeding during gestation and lactation. The offspring whose dams were fed with HFD had a lower birth weight but an increased body weight with impaired glucose tolerance, higher serum cholesterol, and hepatic steatosis at weaning. Microarray analyses showed that there were 120 genes differently expressed between the two groups. We further verified the results by qRT-PCR. Significant increase of the lipogenesis ( Me1 , Scd1 ) gene expression was found in HFD ( P <0.05), and up-regulated expression of genes ( PPAR-α , Cpt1α , Ehhadh ) involved in β-oxidation was also observed ( P <0.05), but the Acsl3 gene was down-regulated ( P <0.05). Maternal over-nutrition could not only primarily induce lipogenesis, but also promote lipolysis through an oxidation pathway as compensation, eventually leading to an increased body weight, impaired glucose tolerance, elevated serum cholesterol and hepatic steatosis at weaning. This finding may provide some evidence for a healthy maternal diet in order to reduce the risk of metabolic diseases in the early life of the offspring.

This study aimed to assess the impact of perinatal high-fat (HF) diet in female Sprague-Dawley rats (F0) on glucose metabolism and islet function in their early life of second-generation of offspring (F2).

Objective To investigate effect of resveratrol on ventricular arrhythmias in rats with myo-cardial infarction .Methods Twenty-four adult male Sprague-Dawley rats were randomly divided into myocar-dial infarction group(MI group,n=8), resveratrol group (Res group,n=8) after successful establishment of MI model,and sham group (n=8) which underwent thoracotomy and pericardiotomy without the left anterior descending ligation.Res group was given a dose of 10 mg? kg-1? d-1 resveratrol (i.g.once a day) for 4 weeks. MI group and sham group received 5%sodium carboxymethyl cellulose gavage .At 4 weeks after induced MI the programmed electrical stimulation was performed in vivo electrophysiological studies .Myocardial infarct size was detected by Masson’s staining.The distribution of connexin43(Cx43) and P368Cx43 were detected by immuno-histochemical staining methods .The Cx43,P368Cx43 and PKCβ2 expressions were measured using western blot . Results Compared with MI group ,resveratrol significantly reduced the incidence rate of induced ventricular arrhythmias (87.5%vs.37.5%,P<0.05),and the ventricular fibrillation threshold was significantly higher [(3.2±0.3)V vs.(5.2±0.8)V,P<0.05].Compared with the sham group,the expression of Cx43 within the infracted myocardium was significantly lower (P<0.05),while the expressions of P368Cx43 and PKCβ2 were greatly higher ( P<0.05 ) .Cx43 levels increased in myocardial infarction rats which received resveratrol ( P<0.05).Conclusion Resveratrol could effectively improve the cardiac electrophysiological properties in MI rats and increase myocardial expression of Cx 43 protein, and could prevent the occurrence of ventricular arrhythmias.

目的 研究白藜芦醇(RSV)对脂多糖(LPS)诱导的H9c2细胞损伤的保护作用及其机制.方法 用LPS 10 μg/mL处理大鼠H9c2心肌细胞0、5、10、20、30、60 min,评估LPS在不同时间点对H9c2细胞核因子κB抑制蛋白(IκBα)、p-p65、p65和p-肌动蛋白(β-actin)表达的诱导作用;用不同浓度RSV预处理H9c2细胞后再用LPS刺激,观察RSV对LPS诱导的H9c2细胞损伤的影响.MTT法检测H9c2细胞增殖能力;荧光法检测细胞内活性氧(ROS)水平;Western blotting检测IκBα、p-p65、p65和β-actin的表达.结果 LPS抑制H9c2细胞增殖,使细胞内ROS产生量明显增加;LPS使IκBα表达水平显著降低,使p65磷酸化水平显著升高.RSV预处理能明显降低LPS对H9c2细胞的损伤,使细胞内ROS产生明显减少;RSV可上调IκBα表达,下调p-p65的表达.结论 RSV可能通过下调p-p65及上调IκBα抑制LPS诱导的H9c2细胞损伤.

Objective To study the impact of maternal high-fat diet during pregnancy and lactation on hepatic steatosis in the early life of offspring rats and its possible mechanism. Methods Female Sprague-Dawley rats were fed either a high fat diet (HF) or control (C) diet for 8 weeks before mating and throughout gestation and ifrst 3 weeks of lactation. The expressions of hepatic fatty acid catabolism related genes, including peroxisome proliferator-activated receptor alpha (PPARα), acyl-CoA syn-thease long-chain family member3 (ACSL3), carnitine palmitoyltransferase-1α(CPT-1α) and 3-hydroxyacyl CoA dehydrogenase (Ehhadh) were determined in offspring liver tissue. The liver pathology was examined in offspring rats at 3 weeks of age. Results Pathohistological ifndings at 3 weeks of age showed that there were diffuse vacuolar degeneration in cytoplasm of hepatocytes and spot necrosis in hepatic lobular in the HF offspring liver. The mRNA expressions of PPARαand Ehhadh genes were markedly increased in the HF offspring as compared to the control group (P<0.05). The mRNA expression of CPT-1αgene was also higher in the HF offspring than that in control group (P=0.19). The level of ACSL3 gene expression, however, was markedly decreased (P<0.05). Conclusions Maternal high fat diet during pregnancy and lactation could result in an increased expression of genes related to hepatic fatty acidβ-oxidation, including PPARα, CPT1αand Ehhadh, but the liver steatosis cannot be reversed in the early life of offspring.

Objective To investigate whether resveratrol ( RSV) protects H9c2 cells against lipopolysaccharide ( LPS) induced oxi-dative injury partly through ERK1/2 signaling pathway.Methods H9c2 cells were divided into six groups:control, LPS (10μg/ml li-popolysaccharide),L+R5(10μg/ml lipopolysaccharide +5 μmol/L resveratrol),L+R10(10μg/ml lipopolysaccharide +10μmol/L res-veratrol),L+R20(10μg/ml lipopolysaccharide +20 μmol/L resveratrol),L +R50(10μg/ml lipopolysaccharide +50μmol/L resvera-trol).H9c2 cells from group L+R5 ,L+R10,L+R20,L+R50 were pretreated with resveratrol .Then, cells from group LPS ,L+R5,L+R10,L+R20,L+R50 were incubated with LPS for 20 min and 12 hours.MTT were used to detect cell proliferation .The protein level of ERK1/2, and phosphorylation of ERK 1/2 was measured by Western blot respectively .Results Compared with control , LPS signifi-cantly reduced cell proliferation (P<0.05), increased(P<0.01)cell death, and up-regulated the level of phosphorylation of ERK 1/2(P<0.01).Pretreatment of resveratrol attenuated the inhibition of LPS on cell viability and down -regulated the level of phosphoryla-tion of ERK1/2.Conclusion Resveratrol may exert its cytoprotection effects on LPS -induced H9c2 cells via down regulating the activa-tion of p-ERK1/2

目的 观察母代高脂饮食对F2代大鼠的不良代谢影响.方法 3周龄SD雌性大鼠随机分为高脂饮食组(F0-HF组)和对照组(F0-CON组),至11周龄时交配怀孕产仔(F1代),母鼠按原饲养要求喂养至哺乳期结束.F1代大鼠断乳后均予普通饲料喂养至11周龄时再次交配产仔(F2-HCC组和F2-CON组),两组F2代大鼠均普通饲料喂养至3周龄,观察其糖脂代谢相关指标变化.结果 F2-HCC组大鼠出生、1周龄时体质量明显高于对照组大鼠,差异有统计学意义(P均＜0.05);F2-HCC组大鼠3周龄时空腹血糖和糖耐量曲线下面积均高于对照组,血清总胆固醇和肝脏三酰甘油水平均高于对照组,差异有统计学意义(P均＜0.05);1日龄及3周龄时F2-HCC组大鼠表现出不同程度的脂肪肝,对照组大鼠肝脏镜下结构均正常.结论 母代高脂饮食可引起F2代大鼠体质量增加、肝脏脂肪变性及糖耐量受损,后代发生代谢性疾病的危险性增加.

目的 探讨糖尿病大鼠发生急性低血糖时,室性心律失常发生的风险及其机制.方法 用链脲佐菌素(STZ)腹腔注射方法制备糖尿病大鼠(DM)模型,将造模成功的大鼠(随机血糖＞ 19.4mmol/L)饲养4周后随机分为糖尿病高血糖组(DM组,n=9)和糖尿病低血糖组(DMHY组,n=9),同时将同一批次正常大鼠随机分为正常血糖组(N组,n=9),正常大鼠低血糖组(NHY组,n=9).DMHY组和NHY组分别给予腹腔注射8U/kg剂量的胰岛素.DM组和N组则给予同体积生理盐水腹腔注射.当血糖＜ 3.0mmol/L时,开胸给予在体电生理刺激.结果 与N组相比,DM组血糖水平明显升高,体重明显降低,心肌重量也明显减少,室性心律失常的诱发率也明显增加.糖尿病4周后与N组相比,DM组QTc间期延长,但差异无统计学意义(P＞0.05);同时诱发的室性快速心律失常(IVT)发生率增高,心室颤动阈值明显降低(9.25±0.56 vs5.56±1.45,P均＜0.05).与N组相比,NHY组K+、肾上腺素及去甲肾上腺素水平差异有统计学意义(P＜0.05).结论 糖尿病大鼠本身室性心律失常发生率明显比正常大鼠高;当发生急性低血糖时,室性心律失常的发生率明显增加,急性低血糖时降低了心室颤动阈值从而促进室性心律失常的发生,血钾降低、肾上腺素及去甲肾上腺素的升高也参与了心律失常的发生.

目的 研究胰岛素诱导的大鼠急性低血糖对心律失常发生及心肌缝隙连接蛋白43(connexin 43,Cx43)表达的影响.方法 链脲佐菌素(STZ)腹腔注射制备糖尿病(DM)大鼠模型.造模成功的大鼠随机分为糖尿病高血糖组(DM组,n=9)和糖尿病低血糖组(DMHY组,n=9);同时将同一批次未建模大鼠随机分为正常血糖组(Sham组,n=9)和低血糖组(NHY组,n=9).DMHY组和NHY组大鼠经颈静脉分别滴注胰岛素诱导低血糖.采用开胸在体电生理刺激诱发心律失常,对各组大鼠血清肾上腺素和去甲肾上腺素浓度进行测定,Western blotting检测各组大鼠心肌Cx43表达.结果 与Sham组比较,DM组大鼠血糖水平升高,体质量降低,室性心律失常的诱发率明显增高(P ＜0.05);NHY组大鼠心律失常诱发率明显增高,室颤阈值显著降低(P＜0.05).与DM组比较,DMHY组心律失常诱发率明显增高,室颤阈值显著降低(P＜0.05).各组心肌Cx43表达量由高到低依此为Sham组、DM组、NHY组和DMHY组,低血糖组与对应的非低血糖组之间的差异均有统计学意义(P＜0.05).与对应的非低血糖组(Sham组和DM组)比较,低血糖组(NHY组和DMHY组)大鼠血清肾上腺素和去甲肾上腺素水平显著升高(P＜0.05).结论 胰岛素诱导的急性低血糖可促进心律失常的发生,其机制可能与室颤阈值降低和心肌Cx43表达减少有关.

目的 探讨细胞内基质金属蛋白酶2(MMP-2)基因过表达对大鼠胰岛β细胞株INS-1细胞功能的影响. 方法 采用基因重组技术将大鼠MMP-2 cDNA插入真核表达载体pcDNA 3.1(+)构建大鼠MMP-2真核表达质粒,培养INS-1细胞.随机分为正常对照组,空质粒转染组及MMP-2质粒转染组.脂质体Lipofectmine 2000转染INS-1细胞,观察INS-1细胞内MMP-2基因和蛋白表达量变化,MMP-2酶活性变化,以及INS-1细胞凋亡情况与胰岛素分泌功能的变化. 结果 与正常对照组、空质粒转染组比较,MMP-2质粒转染组MMP-2 mRNA表达增加(P均＜0.05),蛋白表达水平和酶活性上调(P均＜0.05); MMP-2质粒转染组细胞凋亡率(56.07±3.68)％高于正常对照组(33.70±6.53)％及空质粒转染组(38.02±5.60)％(P＜0.05),而IRI(1.30±0.27)低于正常对照组(3.37±0.76)与空质粒转染组(2.90±0.84) (P＜0.05). 结论 细胞内MMP-2基因的过表达可引起胰岛β细胞凋亡增加,胰岛素分泌功能下降.

OBJECTIVE:To investigate the role of matrix metalloproteinase 2 (MMP2) in pancreatic beta cell injury induced by oxidative stress.METHODS:Rat pancreatic beta cell line INS-1 cells were treated with advanced glycation end-products (AGE) to induce intracellular oxidative stress. Intracellular MMP2 expression and activity were determined by quantitative reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and zymography, respectively. MMP2 expression and activity were manipulated by over-expression with recombinant MMP2 plasmids or knockdown with either MMP2 specific siRNA or inhibitors, and effects on apoptosis and insulin-secretion were measured by flow cytometry and ELISA.RESULTS:AGE treatment induced intracellular oxidative stress in INS-1 cells, as indicated by elevated ROS levels, apoptotic cell death, and suppressed insulin secretion. This was accompanied by increased MMP2 expression and activity. However, Antioxidant N-acetylcysteine (NAC) treatment inhibited MMP2 expression and activity, and partially reversed cell apoptosis and insulin secretion dysfunction induced by AGE. Forced expression of MMP2 mimicked the effects of AGE treatment while inhibition of MMP2 either by a specific MMP2 inhibitor or MMP2 siRNA protected oxidative stress induced by AGE.CONCLUSION:MMP2 expression and intracellular activity are increased by oxidative stress, contributing to cellular dysfunction and apoptosis in INS-1 cells after AGE challenge.

Relevant animal experiments and epidemiological research findings support that in the early development special nutritional status,such as intrauterine growth restricted,maternal low-calorie,low-protein and high-fat diet during gestation,can change their expression of metabolism-related genes by modifying epigenetic abnormally that lead to the development of metabolic diseases in offsprings.Epigenetic modifications mainly include DNA methylation,histone modifications and non-coding RNA regulation.These modifications occurred in early life belong to the category of Developmental Origins of Health and Disease Hypothesis,and its mechanisms,especially its maturing methods on epigenetic research,can provide a reference to guide a healthy diet habit during pregnancy in order to reduce the risk of developing metabolic diseases in their offsprings later.

Objective To investigate the effects of rosiglitazone on vascular lesions in rats with diabetes mellitus,and explore its possible mechanism.MethodsTwenty-four SD rats were randomly divided into normal control group,diabetes mellitus group and diabetes mellitus rosiglitazone treatment group.Rat models of type 1 diabetes mellitus were established in diabetes mellitus group and diabetes mellitus rosiglitazone treatment group by intraperitoneal injection of 60 mg/kg of streptozocin(STZ),and intragastric administration of rosiglitazone(1mg/kg) was conducted in diabetes mellitus rosiglitazone treatment group for 8 weeks from the second week after glucostasis.At the end of the eighth week,heart blood samples were collected from each group of rats,and ELISA was employed to detect the levels of phospholipase A2(PLA2).Besides,thoracoabdominal aorta were obtained,histomorphology and ultrastructure changes of vessel wall were observed,and expression of PLA2 protein of vessel wall was detected by immunohistochemistry.ResultsCompared with normal control group,the vascular structure of rats in diabetes mellitus group was damaged severely,both local PLA2 protein expression and serum PLA2 level increased.The vascular lesions in diabetes mellitus rosiglitazone treatment group were less severe than those in diabetes mellitus group.The expression of PLA2 protein of vessel wall and serum PLA2 level in diabetes mellitus rosiglitazone treatment group were significantly higher than those in control group,and significantly lower than those in diabetes mellitus group(P<0.05).ConclusionRosiglitazone has protective effects on vascular structure of rats with diabetes mellitus,and the mechanism may relate to the inhibition of PLA2 expression and reduction of inflammation.

T2DM组67例(分为非腹型肥胖组、腹型肥胖组、肥胖组)及正常对照组19例,测量FPG、h2PG、CP、GG。结果T2DM患者餐后C肽、GG显著升高[(4.92±4.01vs2.11±0.67)ng/mL,(201.22±46.66vs125.23±40.35)pg/mL,均P<0.01];与非腹型肥胖组相比,腹型肥胖组的T2DM患者餐后C肽及其增加值显著升高[(6.65±4.62vs3.46±2.22)ng/mL,(5.07±4.19vs2.21±1.81)ng/mL,均P<0.05],餐后胰高血糖素及其增加值相对升高但未达到统计学意义[(204.35±43.76vs203.06±51.27)pg/mL,(25.16±36.20vs16.04±19.14)pg/mL,均P>0.05];与腹型肥胖组相比,肥胖组餐后C肽、GG及相应增加值升高,但未达到统计学意义。结论T2DM患者的胰岛α细胞存在胰岛素抵抗,腹型肥胖者胰岛α细胞的胰岛素抵抗更显著。

ed in vitro and in vivo,which was inhibited by aspirin treatment.

Objective To acquire oval cells (progenitor stem cells) from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley (SD) rats were divided into 5 groups randomly: control, 2-acetylaminofluorene (2-AAF), 2-AAF+partial hepatectomy (PH), 2-AAF+carbon tetrachloride (CCl4), and diabetic groups. As two-step collagenase perfusion protocol of Seglen, oval cells were isolated by Percoll density gradient centrifugation. Thy1.1 positive cells were sorted by flow cytometry, and then cultured in Dulbecco's minimum Eagle's medium (DMEM). Immunofluorescence staining was applied to labelling Thy1.1. Results Different rates of Thy1.1 positive oval cells were found in different rat model groups: 0.5% in 2-AAF, 0.3% in 2-AAF+PH, 0.2% in 2-AAF+CCl4 , 0.1% in diabetic, and 0.0% in control. Isolated cells adhered to plate with fusiform or polygon as epithelial cells. Conclusion Progenitor stem cells exist in injured liver tissue including those from diabetic rats.

Objective To observe the effect of Aspirin,prednisone and Acertil on expression of cyclooxygenase-2 mRNA in the abdominal aorta of diabetes mellitus (DM) rats.Methods Twenty-five Sprague-Dawely rats were randomly divided into normal control group,DM group,Aspirin group(Aspirin,1 mg·kg-1·d-1),prednisone group (prednisone,1 mg·kg-1·d-1) and Acertil group (Acertil,1 mg·kg-1·d-1). All rats were sacrificed ten weeks later and the expression of cyclooxygenase-2 was investigated with reverse transcriptase-polymerase chain reaction (PCR) and real time-PCR. Results The expression of cyclooxygenase-2 mRNA was observed at 228 bp site in all the groups as expected,with the strongest expression observed in the DM group,followed by Acertil group,Aspirin group,prednisone group,and normal control group in order. Real-time PCR showed that the relative amount of COX-2 mRNA expression was 0.29±0.02 in the normal control group,significantly lower than that in other groups (P0.05,or 0.01); that in the DM group (0.83±0.02) was significantly higher than those in the Acertil group (0.59±0.02),Aspirin group (0.56±0.13) and prednisone group (0.38±0.06,all P0.01); and that in the Acertil group was significantly higher than that in the prednisone group (P0.05). Conclusion The expression of cyclooxygenase-2 can be taken as an index of vascular inflammation of diabetes mellitus. Inhibition of cyclooxygenase-2 might be a new therapeutic method for vascular inflammation in diabetic patients. Prednisone,Aspirin and Acertil can lessen vascular inflammation in diabetes,with prednisone being the most effective one.

Objective To acquire oval cells （progenitor stem cells） from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley （SD） rats were divided into 5 groups randomly: control, 2-acetylaminofluorene （2-AAF）, 2-AAF+partial hepatectomy （PH）, 2-AAF+carbon tetrachloride （CCl4）, and diabetic groups. As two-step collagenase perfusion protocol of Seglen, oval cells were isolated by Percoll density gradient centrifugation. Thy1.1 positive cells were sorted by flow cytometry, and then cultured in Dulbecco’s minimum Eagle’s medium （DMEM）. Immunofluorescence staining was applied to labelling Thy1.1. Results Different rates of Thy1.1 positive oval cells were found in different rat model groups: 0.5% in 2-AAF, 0.3% in 2-AAF+PH, 0.2% in 2-AAF+CCl4 , 0.1% in diabetic, and 0.0% in control. Isolated cells adhered to plate with fusiform or polygon as epithelial cells. Conclusion Progenitor stem cells exist in injured liver tissue including those from diabetic rats.

The response of glueagon to the change of glucose and its relation to blood pressure in 71 eidely patients with type 2 diabetes was investigated. The results showed that the postprandial increment of glucagon in the group of patients with postprandial 2h plasma glucose increment<2.5 mmol/L was significantly higher than that in the group with plasma glucose inerement≥ 2.5 mmoi/L (P<0. 05). The postprandial increment of glucagon in patients with normal blood pressure was significantly higher than that in patients with hypertension (P<0.05). The results suggest that the decreased response of glucagon to the change in plasma glucose in elderly patients with type 2 diabetes is related to increased blood glucose and high blood pressure.

Objective To observe the structure changes of liver in diabetic rats and the influence of rosiglitazone on the ultrastructure of hepatocytes.Methods Thirty Sprague Dawley(SD)rats were divided into 3 groups:control group(C),receiving peritoneal injection of 0.1 mmol/L(pH 4.5)sodium citrate solution;diabetes group(D),receiving peritoneal injection of streptozocin(STZ,60 mg/kg)+0.1 mmol/L(pH 4.5)sodium citrate solution;and diabetes plus rosiglitazone treatment group(DR),receiving normal saline and rosiglitazone(ig,1 mg·kg-1·d-1)3 days after treatment as in group D.After 10 weeks,the liver tissues were observed under the transmission electron microscope and two independent pathologists calculated the scores of damage.Results Hepatopathy was observed in group D,with swelling,atrophic mitochondria and outstretched cholangiole of the central zone.The score of damage degree was 23.9±0.8 in group D,which was significantly higher than those of the control group(4.7±0.2,P=0.001)and DR group(5.2±0.1,P=0.001).Conclusion Rosiglitazone can protect hepatocyte and cholangiole in diabetic rats induced by STZ.(Shanghai Med J,2008,31:785-787)