益生菌对胆汁淤积性黄疸大鼠小肠上皮细胞紧密连接蛋白的影响

目的探讨双歧三联活菌(培菲康)对胆汁淤积性大鼠小肠上皮细胞紧密连接蛋白ZO-1(zonula oc-cludens-1)和Occludin表达的调控机制。方法雄性3周龄SD大鼠随机分为对照组、模型组和培菲康组,模型组和培菲康组均给予α-异硫氰酸萘酯(ANIT)50 mg/kg一次性灌胃,建立急性肝内胆汁淤积动物模型,培菲康组于造模前4 d开始给予培菲康4.2×107个活菌数/(kg.d)灌胃大鼠。分别于造模后24、48和72 h三个时间点处死大鼠,取末端回肠黏膜组织,采用免疫组化和Western blots免疫印迹法检测紧密连接蛋白ZO-1、闭锁蛋白(Occludin)的分布和表达,并利用图像分析系统对Western blots图像结果进行定量分析。结果ZO-1和Occludin蛋白主要沿大鼠小肠黏膜上皮细胞膜的顶端呈线状分布,模型组大鼠24 h时ZO-1和Occludin的阳性染色较对照组减少,48 h减少最为明显,72 h阳性染色有所恢复,而培菲康组大鼠各时间点ZO-1和Occludin的阳性染色和模型组相比均明显增多。Western blots结果与免疫组织化学结果相一致,模型组24 h已经开始下降(ZO-1 0.1294±0.0481)、(Oc-cludin 0.1950±0.0441),48 h达到最低(ZO-1 0.0395±0.0095)、(Occludin 0.0137±0.0092),72 h开始恢复(ZO-10.2024±0.0498)、(Occludin 0.1494±0.0355),各时间点与对照组(ZO-1 0.2887±0.0237)、(Occludin 0.4266±0.0670)相比差异有统计学意义(P0.01);而培菲康组各时间点蛋白表达分别为24 h(ZO-1 0.2110±0.0367)、(Occludin 0.3056±0.0572),48 h(ZO-1 0.1173±0.0423)、(Occludin 0.0521±0.0123),72 h(ZO-1 0.2601±0.0191)、(Occludin 0.2050±0.0721),与模型组相应时间点数据相比差异有统计学意义(P0.05)。结论双歧三联活菌能够影响胆汁淤积性大鼠小肠黏膜上皮紧密连接蛋白的分布和表达,可以恢复肠黏膜上皮屏障的完整性。     

二氮嗪对大鼠脑缺血再灌注后ZO-1和Claudin-5蛋白表达的影响

目的观察二氮嗪对大鼠脑缺血再灌注后紧密连接相关蛋白ZO-1和Claudin-5蛋白表达的影响,研究其对血脑屏障紧密连接是否具有保护作用。方法将30只雄性Wistar大鼠随机分为3组:假手术组、缺血再灌注组及二氮嗪预处理组,采用线栓法建立大鼠大脑中动脉缺血再灌注模型,分别应用免疫组化染色和Western blot检测各组大鼠ZO-1和Claudin-5蛋白的表达水平。结果 1.假手术组ZO-1、Claudin-5在血管上的染色连续、丰富,缺血再灌注组ZO-1、Claudin-5的染色稀少、断续,二氮嗪预处理组染色较缺血再灌注组有所改善;2.Western blot定量测定与假手术组相比,缺血再灌注组大鼠脑组织中ZO-1和Claudin-5蛋白表达均明显下降(P0.01),与缺血再灌注组相比,二氮嗪预处理组ZO-1和Claudin-5的蛋白表达明显增多(P0.05)。结论二氮嗪对血脑屏障紧密连接具有保护作用,其机制可能与增加紧密连接相关蛋白ZO-1和Claudin-5的表达有关。     

整肠生对溃疡性结肠炎 小鼠肠上皮屏障的保护作用

目的探讨整肠生对溃疡性结肠炎小鼠肠道紧密连接蛋白表达以及对氧化应激反应的影响。方法选用雄性8~10周龄C57BL/6小鼠40只,随机分为4组:对照组、模型组(3%DSS)、5-ASA组(3%DSS+5-ASA 200mg/kg灌胃)和整肠生组(3%DSS+联合整肠生及5-ASA灌胃),每组10只,造模7d。观察各组小鼠便血程度、组织学损伤情况,通过投射电镜观察各组肠道上皮间紧密连接改变情况,应用Western blot和RT-PCR的方法,检测小鼠结肠黏膜紧密连接蛋白Occludin、ZO-1、Claudin-2的表达情况。结果 (1)与模型组比较,5-ASA组和整肠生组小鼠便血程度明显减轻,DAI评分显著降低(P0.05)。整肠生组与5-ASA组比较,便血减轻,DAI评分降低(P0.05)。(2)电镜显示,对照组肠上皮间紧密连接呈一条致密条带,结构完整,见细胞桥粒,微绒毛光滑、排列整齐,细胞间隙狭窄;模型组肠上皮间紧密连接结构松散、模糊、密度降低,桥粒结构消失,微绒毛稀疏,短缩且长短不一,细胞间隙增宽;各治疗组的紧密连接的破坏情况较模型组有不同程度的改善,整肠生组紧密连接清晰,细胞间隙缩窄,微绒毛排列整齐,出现细胞桥粒。(3)应用Western blot和Real time-PCR法检测,与正常组相比,模型组Occludin、ZO-1蛋白和mRNA表达显著下降,Claudin-2表达显著上调(P0.05);各治疗组较模型组Occludin、ZO-1蛋白表达上调,Claudin-2蛋白表达下调(P0.05),整肠生组较单用5-ASA组更明显提高Occludin、ZO-1蛋白和mRNA表达。(4)与正常组相比,模型组MDA含量增高,SOD活性降低,与5-ASA组相比,整肠生组能更显著地降低MDA含量,提高SOD活性(P0.05)。结论联合应用整肠生通过调节紧密连接蛋白Occludin、ZO-1的表达和降低氧化应激反应,来改善溃疡性结肠炎小鼠肠上皮屏障功能。     

实验性肝硬化大鼠肠上皮细胞间紧密连接蛋白occludin表达下降

目的研究实验性肝硬化大鼠大肠上皮细胞间紧密连接蛋白occludin表达的变化。方法参照文献1,给大鼠反复腹腔注射CCl4制备化学性肝硬化大鼠动物模型。实验4周、8周分批处死动物,应用免疫组织化学及Western blot检测肝硬化进程中,大鼠大肠上皮细胞间紧密连接蛋白occludin的定位及表达的变化。结果occludin蛋白主要沿大鼠大肠粘膜上皮细胞膜的顶端呈线状分布,在肝硬化组大鼠,4周时occludin的阳性染色开始减少,8周时更为明显。Western blot结果与免疫组织化学结果相一致,4周时开始下降(0.51±0.07),8周时达到最低值(0.32±0.05),与对照组(0.83±0.09)相比差异显著(P<0.05)。结论在肝硬化进程中,大肠上皮细胞间紧密连接蛋白occludin表达下降。     

NAFLD患者肠粘膜机械屏障损伤的研究

非酒精性脂肪性肝病(nonalcoholic fatty liver disease,NAFLD)是慢性肝损伤的主要病因之一。据估计,大约有20%的成人有非酒精性脂肪性肝病,2%-3%发展成非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)。NASH是NAFLD的渐进形式,并可能导致肝硬化和肝细胞癌。NAFLD不仅增加了肝病患者死亡率,作为代谢综合征,还增加了肥胖、2型糖尿病及高脂血症的发病率。肌球蛋白轻链激酶(MLCK)是细胞收缩的关键酶,可使肌球蛋白轻链磷酸化(MLC),促使肌动蛋白收缩,破坏细胞间的紧密连接蛋白,使细胞骨架收缩,进而使肠上皮通透性增加,肠粘膜机械屏障遭到破坏,致使NAFLD的病情进一步发展。MLCK在NAFLD的发生及发展中起着重要作用。NAFLD严重威胁人类健康,影响人们的生活质量及生存质量。为NAFLD患者寻找崭新的治疗方法是极其必要的。     

肌球蛋白轻链激酶活性片段对肌球蛋白轻链的非钙依赖性磷酸化作用特征

肌球蛋白轻链激酶 (MLCK)的活性片段 (MLCKF)能比完整的MLCK更有效地、以非钙依赖性的方式磷酸化肌球蛋白轻链 (MLC2 0 )。该片段是用胰蛋白酶水解MLCK ,再经DEAE 5 2柱层析分离而获得的 ,分子量约为 6 1kD。Western印迹已证实该MLCKF与完整的MLCK同源。MLCKF对肌球蛋白轻链的磷酸化作用及其作用特征通过甘油电泳及ScoinImage扫描软件检测 ,肌球蛋白ATP酶活性通过分光光度法检测。实验结果证实 ,MLCKF催化的MLC2 0 非钙依赖性磷酸化 (CIPM)比MLCK催化的CIPM效力高、耗能多 ,但比MLCK催化的MLC2 0 钙依赖性磷酸化 (CDPM)效力低、耗能少 ;MLCKF催化的CIPM与MLCK催化的CIPM均较MLCK催化的CDPM稳定 ,不易受温育温度、温育时间及离子浓度等变化的影响 ,且对MLCK抑制剂ML 9敏感性低。     

肌球蛋白轻链激酶CaM结合位点突变体对肌球蛋白ATP酶活性的影响

目的:探讨肌球蛋白轻链激酶(MLCK)钙调蛋白(CaM)结合位点突变体对肌球蛋白ATP酶活性的影响.方法:构建牛胃重组全长野生型MLCK CaM结合位点突变型蛋白(△CaM/MLCK);孔雀绿方法检测△CaM/MLCK对肌球蛋白的Mg2+-ATP酶活性的影响.结果:在无Ca2+/CaM存在时,随着△△CaM/MLCK浓度的增加,非磷酸化肌球蛋白的Mg2+-ATP酶活性明显增加;而磷酸化肌球蛋白的Mg2+-ATP酶活性明显降低.结论:△CaM/MLCK对肌球蛋白Mg2+-ATP酶活性的影响表明MLCK具有非激酶活性.     

高脂血症对大鼠心电图的影响及其作用机制

目的:探讨在高脂血症状态下,大鼠心电图的变化情况,及高胆固醇血症对心肌电生理特性影响的机制。方法:将20只Wistar大鼠随机分为空白对照组和高脂饮食组,喂养10周后,检测大鼠的血脂水平、心电图和室颤阈值,并通过全细胞膜片钳记录心室肌细胞的ICa,L;利用组织病理学方法评价对照组及高脂饮食组的大鼠动脉粥样硬化的程度。结果:高脂饮食组的大鼠血脂水平与对照组相比明显增高(P<0.01);在高脂饮食组的大鼠动脉血管管壁中,可见广泛分布的粥样硬化斑块。在高脂饮食组的大鼠心电图中,室颤阈值为(4.23±0.12)V,明显低于对照组(12.80±6.34)V,P<0.05。高脂饮食组大鼠的QTc间期(94±16)ms,与对照组(67±12)ms相比明显延长,P<0.05。高脂饮食组大鼠的心室肌细胞的ICa,L密度为(12.83±3.28)pA/pF,与对照组(9.21±2.16)pA/pF相比明显高,P<0.05。结论:高脂饮食后,大鼠的心电图有明显变化,QTc间期延长;高胆固醇血症能明显增加大鼠心肌细胞的ICa,L的,延长复极时程,降低室颤阈值。     

肌球蛋白轻链激酶非激酶活性调节磷酸化肌球蛋白ATP酶活性及肌丝运动

肌球蛋白轻链激酶(myosin light chain kinase,MLCK)具有激酶和非激酶活性,在平滑肌收缩过程中起着关键酶调控的作用.为进一步阐明MLCK非激酶活性在平滑肌收缩过程中的调节作用,利用已删除部分激酶区域的MLCK重组体(pGEXF6.5)在大肠杆菌中进行表达,采用亲和层析技术纯化表达的MLCK片段,应用EnzChek磷分析试剂盒检测MLCK片段对磷酸化肌球蛋白、水解重酶解肌球蛋白(heavymeromyosin,HMM)及肌球蛋白亚片段1(subfragmentl,S1)ATP酶活性的影响,体外检测MLCK片段对肌动蛋白肌丝运动的调节.研究结果显示,pGEX-F6.5重组表达载体在大肠杆菌中以可溶性GST融合蛋白的形式表达.该融合蛋白经Glutathione-Sepharose4B纯化、SDS-PAGE鉴定得到较纯的单一表达条带.纯化的MLCK片段对磷酸化肌球蛋白、HMM和S1的ATP酶活性均有明显激活作用.MLCK片段激活磷酸化肌球蛋白ATP酶活性为:Vmax=(19.426±1.669)倍;Km=(0.486±0.106)μmol/L,MLCK片段对磷酸化HMM和S1的ATP酶活性也有相似的刺激作用.体外肌丝运动研究表明,随着MLCK片段浓度的增加,磷酸化肌球蛋白与肌动蛋白结合的数量不断增加,肌丝运动的速度也随之增加.上述结果表明,MLCK的C端非激酶活性具有调节磷酸化的肌球蛋白ATP酶活性及肌丝运动的作用.     

化瘀通络中药全方及拆方对DN大鼠血液相关指标及肾脏足细胞ZO-1表达的干预作用

通过化瘀通络中药全方及拆方对糖尿病肾病(DN)大鼠的24 h尿蛋白定量(UTP)、血脂、血小板活化指标及肾脏足细胞ZO-1的调节作用,探讨化瘀通络中药全方、拆方的不同作用特点及防治DN的可能作用机制。将健康雄性SD大鼠60只分为正常组(C组)10只,造模组50只。造模组大鼠选择高脂高糖饲料喂养联合腹腔注射链脲佐菌素(STZ,1%)35 mg/kg复制糖尿病(DM)大鼠模型,72 h后检测血糖,将成模大鼠(血糖≥16.7mmol/L)随机分为模型组(M组)、化瘀通络中药全方组(Q组)、通络中药组(T组)、化瘀中药组(H组)。按人/鼠体表面积比率换算等效计量法计算各组大鼠用药量,同时C、M两组大鼠灌以等量饮用水,1次/日,连续16周,分别检测各组大鼠24 h UTP、血脂、血小板活化指标等;采用实时荧光定量(Real-time PCR)、蛋白印迹(Western-Blot)方法检测各组大鼠肾组织足细胞ZO-1的基因及蛋白表达情况。实验结果,与C组比较,其余各组大鼠血糖、24 h UTP明显升高(P0.01),血脂、血小板活化指标出现明显变化(P0.05,P0.01);与M组相比,各中药组24 h UTP均明显减少(P0.01),血脂、血小板相关指标明显改善(P0.05,P0.01);中药各组间比较,Q组部分指标的改善优于T组(P0.05)。与C组相比,其他各组大鼠足细胞裂孔膜蛋白ZO-1表达水平均明显降低(P0.01);与M组相比,各中药组大鼠足细胞裂孔膜蛋白ZO-1表达水平均明显升高(P0.01);与Q组比较,T、H组ZO-1基因和蛋白表达水平均明显降低(P0.05,P0.01)。结果表明化瘀通络中药可以降低DN大鼠24 h UTP,其机制可能与调节脂质代谢紊乱,改善血凝状态,上调足细胞裂孔膜蛋白ZO-1的表达有关,并且,化瘀中药与通络中药联合应用有协同作用,为临床优化中药配伍提供实验依据。     

n-6/n-3 多不饱和脂肪酸营养失衡对小鼠精子发生的影响

目的:探讨n-6/n-3多不饱和脂肪酸营养失衡对小鼠精子发生的影响。方法:健康的30只C57/B6雄鼠随机分为对照组(CON)、高脂组(HF)、花生四烯酸组(HF+AA)。喂食16周,做合笼实验并记录致孕率,通过精子动力分析仪检测小鼠精子活力和数量的变化,用Elisa试剂盒测血清中睾酮和甘油三酯水平。通过病理组织染色观察小鼠睾丸组织的形态学变化。利用Realtime-PCR的方法检测小鼠睾丸中Dazl基因表达水平的变化。结果:高脂组、花生四烯酸组与对照组相比精子活力[(16±0.01;12.33±2.83 vs72.2±12.73)%,P0.001],精子数量[(7.5±1.13;6±0.14 vs 13.87±0.35)million/m L,P0.001],致孕率[(28.57;14.29 vs 78.57)%,P0.001]及血清睾酮含量[(0.35±0.14;0.27±0.07 vs 3.51±0.7)ng/m L,P0.001]均显著性降低。高脂组、花生四烯酸组与对照组相比,血清中甘油三酯的含量显著增高[(0.74±0.04;0.74±0.04 vs 0.45±0.04)mmol/L,P0.001]。病理组织染色观察到花生四烯酸组小鼠睾丸组织出现了明显异形,曲细精管内部的初级精母细胞明显缺失,管腔中从初级精母细胞到精子的发生过程出现了变异,精子的数量也显著性下降。参与精子生成过程中的减数分裂前的有丝分裂增殖期、精原细胞的发育等过程的Dazl基因在高脂组和花生四烯酸组小鼠睾丸中的表达量与对照组相比显著降低(0.87±0.05;0.65±0.03 vs 1.07±0.04,P0.05)。结论:膳食中n-3/n-6多不饱和脂肪酸失衡会导致雄鼠精子发生发育的障碍     

Valproic Acid Treatment Inhibits Hypoxia-Inducible Factor 1α Accumulation and Protects against Burn-Induced Gut Barrier Dysfunction in a Rodent Model

Objective

Burn-induced gut dysfunction plays an important role in the development of sepsis and multiple organ dysfunction. Emerging evidence suggests that hypoxia-inducible factor-1α (HIF-1α) is critical in paracelluar barrier functions via regulating vascular endothelial growth factor (VEGF) and myosin light chain kinase (MLCK) expression. Previous studies have also demonstrated that histone deacetylase inhibitors (HDACIs) can repress HIF-1α. This study aims to examine whether valproic acid (VPA), a HDACI, protects against burn-induced gut barrier dysfunction via repressing HIF-1α-dependent upregulation of VEGF and MLCK expression.

Methods

Rats were subjected to third degree 55% TBSA burns and treated with/ without VPA (300mg/kg). Intestinal barrier dysfunction was evaluated by permeability of intestinal mucosa to fluorescein isothiocyanate (FITC)-dextran and histologic evaluation. Histone acetylation, tight junction protein zonula occludens 1 (ZO-1), VEGF, MLCK and HIF-1α were measured. In addition, CaCO₂ cells were transfected with siRNA directed against HIF-1α and were stimulated with CoCl2 (1mM) for 24 hours with/without VPA (2mM) followed by analysis of HIF-1α, MLCK, VEGF and ZO-1.

Results

Burn insults resulted in a significant increase in intestinal permeability and mucosal damage, accompanied by a significant reduction in histone acetylation, ZO-1, upregulation of VEGF, MLCK expression, and an increase in HIF-1α accumulation. VPA significantly attenuated the increase in intestinal permeability, mucosa damage, histone deacetylation and changes in ZO-1 expression. VPA also attenuated the increased VEGF, MLCK and HIF-1α protein levels. VPA reduced HIF-1α, MLCK and VEGF production and prevented ZO-1 loss in CoCl2-stimulated Caco-2 cells. Moreover, transfection of siRNA directed against HIF-1α led to inhibition of MLCK and VEGF production, accompanied by upregulation of ZO-1.

Conclusions

These results indicate that VPA can protect against burn-induced gut barrier dysfunction. These protective effects may be due to its inhibitory action on HIF-1α, leading to a reduction in intestinal VEGF and MLCK expression and minimizing ZO-1 degradation.     

27-nt miRNA对血管内皮细胞eNOS表达/活性的调节及其代谢产物的影响

目的:前期工作表明,27-nt miRNA对eNOS的转录和表达有负反馈调节作用。本实验进一步探讨27-nt miRNA对血管内皮细胞eNOS的基因表达、活性调节及其代谢产物的影响。方法:构建27-nt miRNA高表达质粒,并将其转染至HUVECs。MTT法检测细胞的增殖情况,划痕实验检测细胞的迁移能力,Western Blot检测27-nt miRNA及细胞eNOS蛋白的表达情况,ELISA法检测eNOS活性,硝酸还原法测定细胞培养上清液中NO的含量。结果:27-nt miRNA对HUVECs的增殖有强烈的抑制作用(0.674±0.093 vs 0.315±0.013,0.743±0.076 vs 0.315±0.013,P0.05);27-nt miRNA对HUVECs的迁移有显著的抑制作用(0.483±0.009vs 0.806±0.017,0.465±0.047 vs 0.806±0.017,P0.05);27-nt miRNA显著降低eNOS蛋白的表达(0.410±0.004 vs 0.645±0.007,0.483±0.009 vs 0.645±0.007,P0.05)及明显抑制eNOS的活性(1.093±0.357 vs 5.034±0.509,1.707±0.652 vs 5.034±0.509,P0.05);27-nt miRNA明显抑制NO的合成与释放(70.687±4.432 vs 136.803±6.913,75.264±4.481 vs 136.803±6.913,P0.05)。结论:27-nt miRNA高表达明显抑制eNOS基因的表达及活性;27-nt miRNA明显抑制NO的合成和释放,可能成为血管性疾病治疗的分子靶点。     

miR-125a-5p转染对肝癌细胞增殖、侵袭、迁移的影响及相关机制研究

摘要 目的：探究miR-125a-5p转染对肝癌细胞增殖、侵袭、迁移的影响及相关机制。方法：将肝癌细胞分为对照组、下调组和上调组,并通过细胞转染建立稳定转染的下调组和上调组。MMT法检测细胞增殖能力,流式细胞仪检测细胞凋亡能力,Transwell小室实验检测细胞侵袭能力,细胞划痕实验检测细胞迁移能力,Western blot法检测P13K/Akt通路中AKT、Bax、Bcl-2、P13K、P-AKT蛋白表达量。结果：与上调组相比,下调组24、48、72 h细胞增殖率,细胞侵袭、迁移细胞数,AKT、Bcl-2、P13K、P-AKT蛋白表达量显著降低,具有统计学差异（29.67±9.87 vs 17.34±5.71,t=5.192,P＜0.05、34.75±11.56 vs 15.17±5.04,t=7.365,P＜0.05、38.48±12.81 vs 12.51 ±4.13,t=9.153,P＜0.05,72.53±24.17 vs 36.28±12.07,t=6.365,P＜0.05、86.51±28.75 vs 46.28±15.32,t=5.858,P＜0.05,1.26±0.41 vs 0.81±0.26,t=4.397,P＜0.05、1.35±0.44 vs 0.76±0.24,t=5.584,P＜0.05、1.48±0.46 vs 0.79±0.26,t=6.194,P＜0.05、1.22±0.39 vs 0.73±0.24,t=5.584,P＜0.05）;与上调组相比,下调组24、48、72h细胞凋亡率,Bax蛋白表达量显著升高,具有统计学差异（17.62±5.84 vs 29.31±9.75,t=4.879,P＜0.05、14.97±4.65 vs 34.19±11.36,t=7.427,P＜0.05、11.26±3.74 vs 38.62±12.86,t=9.690,P＜0.05,0.75±0.24 vs 1.33±0.43,t=5.587,P＜0.05）。结论：下调miR-125a-5p的表达,可通过作用于P13K/Akt通路,调控AKT、Bax、Bcl-2、P13K、P-AKT蛋白表达量,进而起到抑制肝癌细胞增殖、促进肝癌细胞凋亡以及抑制肝癌细胞的侵袭、迁移能力。     

Body fat responses to a 1‐year combined exercise training program in male coronary artery disease patients

Objective:

To analyze the body fat (BF) content and distribution modifications in coronary artery disease (CAD) patients in response to a 1‐year combined aerobic and resistance exercise training (CET) program.

Design and Methods:

We followed two groups of CAD male patients for 12 months. One group consisted of 17 subjects (57 ± 12 years) who engaged in a CET program (CET group) and the other was a age‐matched control group of 10 subjects (58 ± 11 years). BF content and distribution were measured through dual energy X‐ray absorptiometry (DXA) at baseline and follow‐up.

Results:

We found no differences on body mass and BMI between baseline and end of follow‐up in both groups but, in CET group, we found significant reductions in all analyzed BF depots, including total BF (21.60 ± 6.00 vs. 20.32 ± 5.89 kg, P < 0.01), % total BF (27.8 ± 5.5 vs. 26.4 ± 5.4%, P < 0.05), trunk fat (12.54 ± 3.99 vs. 11.77 ± 4.01 kg, P < 0.05), % trunk fat (31.1 ± 6.9 and 29.2 ± 7.1%, P < 0.05), appendicular fat (8.22 ± 2.08 vs. 7.72 ± 2.037 kg, P < 0.01), % appendicular fat (25.7 ± 4.9 and 24.5 ± 4.9%, P < 0.05), and abdominal fat (2.95 ± 1.06 vs. 2.75 ± 1.10 kg, P < 0.05). Control group showed significant increase in appendicular fat (7.63 ± 1.92 vs. 8.10 ± 2.12 kg, P < 0.05).

Conclusions:

These results confirm the positive effect of CET on body composition of CAD patients, despite no changes in body mass or BMI. In this study, we observed no alterations on BF distribution meaning similar rate of fat loss in all analyzed BF depots. These results also alert for the limitations of BMI for tracking body composition changes.     

Altered cellular redox status,sirtuin abundance and clock gene expression in a mouse model of developmentally primed NASH

BackgroundWe have previously shown that high fat (HF) feeding during pregnancy primes the development of non-alcoholic steatohepatits (NASH) in the adult offspring. However, the underlying mechanisms are unclear.AimsSince the endogenous molecular clock can regulate hepatic lipid metabolism, we investigated whether exposure to a HF diet during development could alter hepatic clock gene expression and contribute to NASH onset in later life.MethodsFemale mice were fed either a control (C, 7% kcal fat) or HF (45% kcal fat) diet. Offspring were fed either a C or HF diet resulting in four offspring groups: C/C, C/HF, HF/C and HF/HF. NAFLD progression, cellular redox status, sirtuin expression (Sirt1, Sirt3), and the expression of core clock genes (Clock, Bmal1, Per2, Cry2) and clock-controlled genes involved in lipid metabolism (Rev-Erbα, Rev-Erbβ, RORα, and Srebp1c) were measured in offspring livers.ResultsOffspring fed a HF diet developed NAFLD. However HF fed offspring of mothers fed a HF diet developed NASH, coupled with significantly reduced NAD⁺/NADH (p < 0.05, HF/HF vs C/C), Sirt1 (p < 0.001, HF/HF vs C/C), Sirt3 (p < 0.01, HF/HF vs C/C), perturbed clock gene expression, and elevated expression of genes involved lipid metabolism, such as Srebp1c (p < 0.05, C/HF and HF/HF vs C/C).ConclusionOur results suggest that exposure to excess dietary fat during early and post-natal life increases the susceptibility to develop NASH in adulthood, involving altered cellular redox status, reduced sirtuin abundance, and desynchronized clock gene expression.     

烟酰胺核糖抑制db/db小鼠糖尿病心肌病的作用及机制研究

目的:探讨烟酰胺核糖(NR)对2型糖尿病小鼠心肌病的治疗作用及其机制。方法:2型糖尿病模型db/db鼠和及其严格对照小鼠db/+小鼠,将小鼠分为Con (db/+)组,DM (db/db)组,DM+NR组。采用超声测小鼠心脏功能,western-blot及免疫组化测SIRT1表达含量,DHE染色、MDA含量和MnSOD活性检测反映氧化应激水平。结果:与对照组相比,db/db小鼠心脏功能显著下降(LVEF:42.3±7.2vs 73.7±10.2, P0.01;LVFS:22.1±4.2vs 42.7±6.9, P0.01),SIRT1表达量显著下调(P0.01)。NR喂养提高SIRT1表达量(P0.01),并有效改善db/db小鼠心脏功能(LVEF:53.1±8.1vs 42.3±7.2, P0.01;LVFS:33.4±6.9vs 22.1±4.2, P0.01)。同时,NR喂养显著降低了db/db小鼠心肌组织的凋亡水平和氧化应激水平(P0.05)。结论:NR有效改善了db/db小鼠的心功能障碍,降低了db/db小鼠的心肌凋亡水平和氧化应激水平,这些作用的发挥可能与NR增加SIRT1的表达量有关。     

早期肠内营养对胃癌根治术患者术后恢复和免疫功能的影响

目的:评价早期肠内营养(EEN)对胃癌根治术患者术后恢复和免疫功能的影响。方法:选入2011年6月～2014年1月在我院行胃癌根治术治疗的患者60例,根据术后营养方式不同分为EEN组和全肠外营养支持(TPN)组,每组30例。比较两组患者机体恢复及免疫功能情况。术后随访3年,观察并记录两组无进展生存率和总生存率。结果:EEN组术后排气时间[(2.46±0.78)d vs(3.85±1.03)d]、排便时间[(4.03±1.17)d vs(5.67±1.23)d]、进流质时间[(5.88±1.30)d vs(7.26±1.59)d]、进半流食时间[(7.94±1.85)d vs(11.01±2.36)d]和住院天数[(14.87±2.56)d vs(17.54±3.30)d]均显著短于TPN组,差异均有统计学意义(P0.05)。术后第1d,两组各项体液免疫指标(Ig A、Ig G、Ig M浓度)和细胞免疫指标(CD3+、CD4+和CD4+/CD8+水平)均显著下降(P0.05),营养支持后逐渐恢复,而EEN组恢复幅度较TPN组大,差异具有统计学意义(P0.05)。EEN组术后并发症总发生率显著低于TPN组(13.33%vs 36.67%,P0.05)。EEN组患者1年、2年和3年无进展生存率和总生存率均稍高于TPN组,但差异无统计学意义(P0.05)。结论:EEN可有效促进胃癌根治术患者的肠功能恢复,缩短住院时间,提高机体免疫功能,降低并发症的发生,值得在临床推荐应用。     

Clinical Profile of Nonalcoholic Fatty Liver Disease Among Young Patients With Type 1 Diabetes Mellitus Seen at a Diabetes Speciality Center in India

ObjectiveTo estimate the prevalence and clinical profile of nonalcoholic fatty liver disease (NAFLD) among young type 1 diabetes mellitus (T1DM) patients at a tertiary care diabetes center in India.MethodsElectronic medical records of T1DM patients (age at first diagnosis of T1DM ≤ 25 years) registered between January 1992 and May 2013 who had undergone ultrasonography and denied history of any alcohol intake (n = 736) were reviewed. NAFLD was diagnosed if there was any degree of fatty liver. Retinopathy was initially assessed by direct and indirect ophthalmoscopy and later by retinal photography. Nephropathy was diagnosed if urine protein excretion was > 500 mg/day, and neuropathy was diagnosed if a patient’s vibration perception threshold on biothesiometry was ≥ 20 V.ResultsA total of 204/736 (27.7%) T1DM patients had NAFLD. Compared to T1DM subjects without NAFLD those with NAFLD had higher body mass index (BMI) (18.9 ± 4.2 vs. 20.2 ± 4.7 kg/m², P < .001), waist circumference (67.9 ± 13.2 vs. 71.9 ± 13.3 cm, P < .05), systolic blood pressure (110 ± 15 vs. 116 ± 18 mm Hg, P < .001) and diastolic blood pressure (72 ± 9 vs. 74 ± 10 mm Hg, P < .05), while fasting blood glucose (201 ± 101 vs. 183 ± 101 mg/dL, P < .05) and alkaline phosphatase (419 [12.5] vs. 315 [15.8], P < .001) levels were lower in patients with T1DM with NAFLD. Multiple logistic regression analysis showed a significant association between NAFLD and retinopathy (odds ratio [OR]: 2.01, 95% confidence interval [CI]: 1.13-3.43; P = .017, after adjusting for sex, duration of diabetes, overweight/obesity, hypertension, fasting plasma glucose, nephropathy, and nephropathy (OR: 1.89, 95% CI: 1.02-3.50; P = .042), after adjusting for sex and fasting plasma glucose.ConclusionsThis study suggests that NAFLD is also seen among T1DM patients and that it has an independent and significant association with retinopathy and nephropathy. (Endocr Pract. 2014;20:1249-1257)