脂联素基因剔除小鼠糖代谢和骨代谢的初步研究

目的:研究前期建立的脂联素基因剔除小鼠模型在基础状态的葡萄糖代谢及骨代谢状态。方法:采用长期追踪野生型和该基因剔除纯合子小鼠的体重和空腹血糖水平,并进行葡萄糖耐量(GTT)和胰岛素耐量(ITT)试验以评价该小鼠的葡萄糖代谢能力。对骨代谢的初步研究采用检测小鼠骨密度(BMD)的方式。结果:发现该基因剔除小鼠在6w时,出现空腹血糖比对照组显著低下的现象,而随着鼠龄的增长,血糖水平又趋于正常。GTT和ITT试验证明该基因剔除小鼠无糖尿病或胰岛素抵抗表型。BMD检测亦显示该小鼠在基础状态下无骨密度低下的现象。结论:脂联素基因剔除小鼠在基础状态下并无显著的糖代谢和骨代谢异常。     

A tailed primers protocol to identify the association of eNOS gene variable number of tandem repeats polymorphism with ischemic stroke in Chinese Han population by capillary electrophoresis

Endothelial nitric oxide synthase (eNOS) plays an important role in mediating endothelium-dependent vasodilatation and antithrombotic action and is thus involved in the development of ischemic stroke (IS). Controversial results regarding the association of eNOS gene variable number of tandem repeats (VNTR) polymorphism with IS have been reported by conventional PCR-polyacrylamide gel electrophoresis methods. We aimed to identify any common association of eNOS gene VNTR polymorphism with IS in Chinese Han population by capillary electrophoresis (CE). The VNTR polymorphism of 27 bp within the eNOS intron-4 was determined by CE with specially designed tailed primers in Chinese Han patients with IS (n = 457) and matched elderly controls without IS (n = 457). Significant differences in BMI, WHR, hypertension, diabetes, smoking, TG, HDL, LDL, LDL, and FBG were observed between cases and controls. The distributions of eNOS VNTR polymorphism were not significantly associated with IS after adjustment for cardiovascular risk factors (OR = 1.18, 95% CI: 0.82–1.69). This finding was consistent with the further meta-analysis in Asians. The meta-analysis in Americans demonstrated that 4a/4b + 4a/4a genotype was significantly associated with IS risk with an OR of 1.54 (95% CI, 1.09–2.17) compared with the 4b/4b genotype. Our data suggests that BMI, WHR, hypertension, diabetes, smoking, TG, LDL, and FBG may increase the risk of IS. However, eNOS VNTR polymorphism may be not an independent major contributor for IS in Chinese Han population. The VNTR polymorphism might be associated with IS in Americans based on meta-analysis.     

内源性Orexin-A调控大鼠胃运动的中枢及外周机制

目的:探讨内源性Orexin-A(OXA)对大鼠胃运动的中枢和外周作用机制。方法:选取成年Wistar大鼠为研究对象,通过禁食诱导大鼠合成内源性OXA。血浆OXA浓度采用放射免疫法测定。实验前大鼠注射OXA受体拮抗剂SB334867,观察内源性OXA的作用。迷走神经切断术用来观察迷走神经的介导作用。胃排空采用分光光度法测量,消化间期胃运动通过在胃窦部植入一应力传感器测量。Orexin前体(PPO)在胃和下丘脑组织的表达,采用蛋白印迹确定。结果:禁食18 h后,血浆OXA水平和PPO蛋白表达显著增加(P0.05),在禁食36 h组达到最高水平(P0.01)。内源性OXA促进胃排空(P0.05),抑制消化间期胃蠕动(P0.05)。外周注射SB334867均能阻断上述胃动力效应(P0.05),但对PPO表达没有影响。迷走神经切断术不能阻断内源性OXA的介导作用(P0.05)。结论:禁食能诱导内源性OXA的合成,内源性OXA能加速胃排空,同时它又抑制消化间期胃蠕动。     

Fasting physiology of polar bears in relation to environmental change and breeding behavior in the Beaufort Sea

We examined the use of the ratio of serum urea to serum creatinine as a physiological biomarker of fasting to monitor temporal patterns in the feeding ecology of polar bears (Ursus maritimus). Blood was collected from 436 polar bears in the eastern Beaufort Sea during April and May of 1985–1986 and 2005–2006. The proportions of polar bears fasting were 9.6% in 1985, 10.5% in 1986, 21.4% in 2005, and 29.3% in 2006. We used stepwise logistic regression analysis to evaluate factors that could influence the binary response variable of fasting or not fasting. Significant predictor variables of fasting were: the 2005 and 2006 capture years, solitary adult male bears, and adult male bears that were accompanying an estrous female. The increased number of polar bears in a physiological fasting state from all sex, age, and reproductive classes in 2005 and 2006 corresponded with broad scale changes in Arctic sea ice composition, which may have affected prey availability. The higher proportion of adult males fasting from all years was attributed to spring breeding behavior.     

Novel modulators for body weight changes induced by fasting and re-feeding in mice

Catch-up weight gain after malnutrition is a risk factor for metabolic syndrome. Here we show that social isolation enhanced fasting-induced weight loss and suppressed weight gain induced by re-feeding for 6 days following a 24-h fast in prepubertal wild-type mice. These effects of social isolation on weight gain were not associated with significant changes in daily average food consumption. Under the same housing condition, genetic deletion of β-endorphin reduced the fasting-induced weight loss and enhanced the re-feeding-induced weight gain in prepubertal mice. These effects of social isolation or genetic deletion of β-endorphin on these weight changes were attenuated and reversed in postpubertal mice. Moreover, genetic deletion of β-endorphin attenuated these effects of social isolation on the catch-up weight gain in prepubertal mice and reversed them in postpubertal mice. Thus, social isolation, endogenous β-endorphin, and age can be novel modulators for body weight changes induced by fasting and re-feeding in mice.     

Effect of endurance flight on haematocrit in migrating birds

The effects of an endurance flight on the haematocrit, the percentage of packed red blood cells per blood volume, were examined within the framework of six possible factors explaining possible changes in the haematocrit. Two approaches were adopted: (1) the haematocrit was studied in four species of passerine birds which landed on an Italian island after having crossed the Mediterranean Sea on their spring migration in a non-stop flight; (2) the haematocrit was evaluated in six individual red knots after a flight of 1, 2, 4 and 10 h in a wind tunnel and the data thus obtained compared with data on resting birds with or without food. In the four passerine species, the haematocrit decreased from 51% in fat birds to 48% in lean birds. In lean birds, the haematocrit dropped from 48% in birds with well-developed breast muscles to 36% in birds with emaciated breast muscles. In the red knots, the haematocrit was dependent on body mass in flying and resting birds. The haematocrit decreased from about 51% pre-flight to about 49% within 1 h of flight and remained at this level for up to 10 h of flight. Taking the results from the passerines and the red knots together, it seems that the haematocrit drops by a few percentage points within 1 h after the onset of flight, decreases very slowly with decreasing body mass and decreases more steeply in very lean birds having entered stage III of fasting. This indicates that dehydration is not an underlying factor in decreased haematocrit because if this were the case we would expect an increase with endurance flight. We found no effect of the presence of blood parasites on haematocrit. With the onset of flight, haemodilution may be adaptive, because it reduces blood viscosity and, thereby, energy expenditure by the heart, or it may be a sign of water conservation as an insurance against the risk of dehydration during long non-stop flights. During endurance flight, a reduction in the haematocrit may be adaptive, in that oxygen delivery capacity is adjusted to the decreased oxygen needs as body mass decreases. A decreasing haematocrit would also allow birds to reduce heart beat frequency and/or heart size, because blood viscosity decreases disproportionally with decreasing haematocrit. However, when energy stores are about to come to an end and birds increase protein breakdown, the haematocrit decreases even further, and birds probably become anaemic due to a reduced erythropoiesis.     

ASSOCIATIONS OF METABOLIC PARAMETERS AND ETHANOL CONSUMPTION WITH MESSENGER RNA EXPRESSION OF CLOCK GENES IN HEALTHY MEN

Recent studies suggest that the impairment of circadian clock function causes various pathological conditions, such as obesity, diabetes, and alcoholism, and an altered mRNA expression of clock genes was found under these conditions. However, it remains to be determined whether clock gene expression varies depending on metabolic conditions even in healthy people. To address this issue, we investigated the associations of metabolic parameters and alcohol consumption with mRNA expression of clock genes (CLOCK, BMAL1, PER1, PER2, and PER3) in peripheral blood cells obtained from 29 healthy non-obese elderly men (age 51–78 yrs) who adhered to a regular sleep-wake routine, through a single time-of-day venous blood sampling at ～09:00?h. There were significant correlations between (1) waist circumference and mRNA level of PER1 (r?=?0.43), (2) plasma glucose concentration and PER2 (r?=?0.50), (3) ethanol consumption and BMAL1 (r?=?0.43), and (4) serum γ-GTP concentration (a sensitive marker of alcohol consumption) and PER2 (r?=?0.40). These results suggest mRNA expression of clock genes is associated with obesity, glucose tolerance, and ethanol consumption even in healthy people. (Author correspondence: akiofuji@jichi.ac.jp)     

Effects of Ramadan upon Fluid and Food Intake,Fatigue, and Physical,Mental, and Social Activities: A Comparison between the UK and Libya

Two studies were performed during Ramadan, one in the UK (N=31) and the other in Libya (N=33). The aims were to assess some changes to lifestyle that are produced by fasting as well as effects due to culture. Subjects were studied on eight separate occasions: four control days (two before and two after Ramadan) and four days during the four weeks of Ramadan itself. A questionnaire was answered that asked about naps and fluid and food intake. The questions elicited if an individual had slept, drank, or eaten, plus the reasons for doing or not doing so. Also, subjects were asked to describe their physical, mental, and social activities, their fatigue, and their perceived abilities to perform physical or mental work. The questionnaire was answered five times per day: at sunrise, at 10:00 h, at 14:00 h, at sunset, and on retiring to sleep at night. Urine samples were collected at sunset and measured for osmolality. Differences between control and Ramadan days, as well as between subjects studied in UK and Libya, were assessed by analysis of variance. Correlations between fatigue and physical, mental, and social activities were also assessed, as were differences in urine osmolality. Fasting during Ramadan resulted in fewer activities and increased fatigue and frequency of napping during daytime. Changes in fluid and food intake indicated some degree of preparation for fasting before sunrise and a marked “recuperation” from fasting after sunset. The reasons given for napping in the daytime, for drinking or not drinking, and for eating or not eating, changed during Ramadan compared with control days; as a result, links between fatigue and activities, and fatigue and fluid and food intake, were all altered during Ramadan, particularly after sunset. Subjects become dehydrated during the daytime, but this was not reduced when females who were menstruating drank during this time. Several differences between the two studies were found. There was a greater frequency of napping during the daytime in the Libya study, and evidence for the conservation of energy during the daytime and reduced physical, mental, and social activities. Subjects' preparations for fasting and recovering from it—their fluid and food intakes and associated reasons for these—also differed. Possible explanations of these differences are discussed. (Author correspondence: waterhouseathome@hotmail.com)     

Food-Entrained Feeding and Locomotor Circadian Rhythms in Rats Under Different Lighting Conditions

It has been suggested that two endogenous timekeeping systems, a light-entrainable pacemaker (LEP) and a food-entrainable pacemaker (FEP), control circadian rhythms. To understand the function and interaction between these two mechanisms better, we studied two behavioral circadian rhythmicities, feeding and locomotor activity, in rats exposed to two conflicting zeitgebers, food restriction and light-dark cycles. For this, the food approaches and wheel-running activity of rats kept under light-dark (LD) 12:12, constant darkness (DD), or constant light (LL) conditions and subjected to different scheduled feeding patterns were continuously recorded. To facilitate comparison of the results obtained under the different lighting conditions, the period of the feeding cycles was set in all three cases about Ih less than the light-entrained or free-running circadian rhythms. The results showed that, depending on the lighting conditions, some components of the feeding and wheel-running circadian rhythms could be entrained by food pulses, while others retained their free-running or light-entrained state. Under LD, food pulses had little influence on the light-entrained feeding and loco-motor rhythms. Under DD, relative coordination between free-running and food-associated rhythms may appear. In both cases, the feeding activity associated with the food pulses could be divided into a prominent phase-dependent peak of activity within the period of food availability and another afterward. Wheel-running activity mainly followed the food pulses. Under LL conditions, the food-entrained activity consisted mainly of feeding and wheel-running anticipatory activity. The results provide new evidence that lighting conditions influence the establishment and persistence of food-entrained circadian rhythms in rats. The existence of two coupled pacemakers, LEP and FEP, or a multioscillatory LEP may both explain our experimental results.     

The PNPLA3 Ile148Met interacts with overweight and dietary intakes on fasting triglyceride levels

The Ile148Met (rs738409, G-allele) in the patatin-like phospholipase domain-containing protein 3 gene (PNPLA3) associates with liver fat content and may lead to loss-of-function (hydrolysis) or gain-of-function (CoA-dependent lysophosphatidic acid acyltransferase) defects. PNPLA3 is up-regulated by dietary carbohydrates, and interactions between rs738409 and carbohydrates, and sugar and ω6:ω3-polyunsaturated fatty acid (PUFA) ratio on hepatic fat accumulation have been reported. We examined interaction between rs738409 and overweight, and between rs738409 and dietary intakes (carbohydrates, sucrose and ω6:ω3-PUFA ratio), on fasting triglyceride levels. From the Malmo Diet and Cancer Study-Cardiovascular Cohort, 4,827 individuals without diabetes aged 58 ± 6 years, 2,346 with BMI ≤ 25 kg/m² and 2,478 with BMI > 25 kg/m², were included in cross-sectional analyses. Dietary data were collected by a modified diet history method. Overweight modified the association between rs738409 and fasting triglyceride levels (P_interaction = 0.003). G-allele associated with lower triglycerides only among overweight individuals (P = 0.01). Nominally, significant interaction on triglyceride levels was observed between rs738409 and sucrose among normal-weight individuals (P_interaction = 0.03). G-allele associated with lower triglycerides among overweight individuals in the lowest tertiles of carbohydrate and ω6:ω3-PUFA ratio (P = 0.04 and P = 0.001) and with higher triglycerides among normal-weight individuals in the highest tertile of sucrose (P = 0.001). We conclude that overweight and dietary sucrose may modify the association between rs738409 and fasting triglyceride levels.

Electronic supplementary material

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