The effect of food deprivation and experimental diabetes on orexin and NPY mRNA levels

Although exogenous orexin can induce feeding, reports of increased orexin gene expression after caloric manipulations have been inconsistent. We hypothesized that orexin gene expression is increased only by extreme negative energy balance challenges. We measured hypothalamic orexin and NPY mRNA by in situ hybridization and orexin-A immunoreactivity in rats after food deprivation, streptozotocin-induced diabetes, and combined deprivation and diabetes. Neither food deprivation, nor diabetes, nor the combination affected orexin mRNA levels, although orexin-A immunoreactivity was increased by diabetes. NPY mRNA levels were increased by either treatment. These results suggest that increased orexin gene expression is not a consistent correlate of negative energy balance challenges.     

MCH levels in the CSF,brain preproMCH and MCHR1 gene expression during paradoxical sleep deprivation,sleep rebound and chronic sleep restriction

Neurons that utilize melanin-concentrating hormone (MCH) as neuromodulator are located in the lateral hypothalamus and incerto-hypothalamic area. These neurons project throughout the central nervous system and play a role in sleep regulation. With the hypothesis that the MCHergic system function would be modified by the time of the day as well as by disruptions of the sleep-wake cycle, we quantified in rats the concentration of MCH in the cerebrospinal fluid (CSF), the expression of the MCH precursor (Pmch) gene in the hypothalamus, and the expression of the MCH receptor 1 (Mchr1) gene in the frontal cortex and hippocampus. These analyses were performed during paradoxical sleep deprivation (by a modified multiple platform technique), paradoxical sleep rebound and chronic sleep restriction, both at the end of the active (dark) phase (lights were turned on at Zeitgeber time zero, ZT0) and during the inactive (light) phase (ZT8).We observed that in control condition (waking and sleep ad libitum), Mchr1 gene expression was larger at ZT8 (when sleep predominates) than at ZT0, both in frontal cortex and hippocampus.In addition, compared to control, disturbances of the sleep–wake cycle produced the following effects: paradoxical sleep deprivation for 96 and 120 h reduced the expression of Mchr1 gene in frontal cortex at ZT0. Sleep rebound that followed 96 h of paradoxical sleep deprivation increased the MCH concentration in the CSF also at ZT0. Twenty-one days of sleep restriction produced a significant increment in MCH CSF levels at ZT8. Finally, sleep disruptions unveiled day/night differences in MCH CSF levels and in Pmch gene expression that were not observed in control (undisturbed) conditions.In conclusion, the time of the day and sleep disruptions produced subtle modifications in the physiology of the MCHergic system.     

大鼠室旁核注射orexin-A对体重的影响

目的:探讨大鼠室旁核(PVN)注射orexin-A对体重的影响。方法:大鼠室旁核(PVN)微量注射orexin-A,用大脑置管埋管、组织化学染色等方法探讨PVN注射orexin-A对其体重的影响。结果:与安慰剂组大鼠相比,PVN注射orexin-A组大鼠体重明显减轻(P0.05),而orexin-A组和安慰剂组摄食量无明显差异(P0.05)。注射结束后6天,orexin-A处理大鼠的体重仍显著低于注射前(P0.05),而安慰剂组大鼠则比注射前显著增重(P0.05)。药物注射可显著降低机体脂肪,但并不特异存在于注射orexin-A或安慰剂的大鼠身上。Orexin-A组和安慰剂组大鼠的肌肉量和脂肪量均显著降低(P0.05),但注射orexin-A的大鼠降低更明显。与安慰剂组相比,orexin-A处理后的摄食转化率显著降低(P0.05)。结论:大鼠室旁核(PVN)注射orexin-A可通过增加活动量产生负能量平衡,引起体重减轻。     

Spinal neuropeptide expression and neuropathic behavior in the acute and chronic phases after spinal cord injury: Effects of progesterone administration

Patients with spinal cord injury (SCI) develop chronic pain that severely compromises their quality of life. We have previously reported that progesterone (PG), a neuroprotective steroid, could offer a promising therapeutic strategy for neuropathic pain. In the present study, we explored temporal changes in the expression of the neuropeptides galanin and tyrosine (NPY) and their receptors (GalR1 and GalR2; Y1R and Y2R, respectively) in the injured spinal cord and evaluated the impact of PG administration on both neuropeptide systems and neuropathic behavior. Male rats were subjected to spinal cord hemisection at T13 level, received daily subcutaneous injections of PG or vehicle, and were evaluated for signs of mechanical and thermal allodynia. Real time PCR was used to determine relative mRNA levels of neuropeptides and receptors, both in the acute (1 day) and chronic (28 days) phases after injury. A significant increase in Y1R and Y2R expression, as well as a significant downregulation in GalR2 mRNA levels, was observed 1 day after SCI. Interestingly, PG early treatment prevented Y1R upregulation and resulted in lower NPY, Y2R and GalR1 mRNA levels. In the chronic phase, injured rats showed well-established mechanical and cold allodynia and significant increases in galanin, NPY, GalR1 and Y1R mRNAs, while maintaining reduced GalR2 expression. Animals receiving PG treatment showed basal expression levels of galanin, NPY, GalR1 and Y1R, and reduced Y2R mRNA levels. Also, and in line with previously published observations, PG-treated animals did not develop mechanical allodynia and showed reduced sensitivity to cold stimulation. Altogether, we show that SCI leads to considerable changes in the spinal expression of galanin, NPY and their associated receptors, and that early and sustained PG administration prevents them. Moreover, our data suggest the participation of galaninergic and NPYergic systems in the plastic changes associated with SCI-induced neuropathic pain, and further supports the therapeutic potential of PG- or neuropeptide-based therapies to prevent and/or treat chronic pain after central injuries.     

The effect of chronic food and water restriction on open-field behaviour and serum corticosterone levels in rats

In operant conditioning experiments, two methods are commonly used to motivate laboratory rats to perform designated tasks. The first is restricting food so that rats are forced to lose 20% of body weight within one week, followed by maintenance at 80% of the baseline weight for the remainder of the experiment. The second is restricting access to water to 15 min in each 24 h period. These methods are effective in motivating the animals. There is, however, little information available on the effects on performance in tests of behaviour that are not related to operant conditioning. In addition, it is not clear if these commonly used methods of food and water restriction will lead to physiological stress as indicated by an elevation of serum corticosterone. Male rats were either food-restricted to reduce and maintain their weight at 80% of baseline weight, or were restricted to 15 min access to water every 24 h. Activity in the open field was significantly greater in food-restricted rats than in water-restricted or control rats, but freezing behaviour was similar in all experimental groups. Food-restricted rats had a higher mean serum corticosterone level than water-restricted and control rats 37 days after the start of the experimental period. These data suggested that chronically restricting food and maintenance of body weight at 80% of baseline body weight led to significant behavioural changes and physiological stress. In contrast, water restriction did not lead to changes in behaviour or corticosterone levels. A second experiment was conducted to compare the effects of food restriction to 80% of baseline body weight, as described above, with a less stringent protocol in which test rats were initially reduced to 80% of baseline weight, but were then maintained at 80% of an ad libitum fed control rat's weight. Serum corticosterone levels and adrenal gland weights were measured after the initial week of forced weight loss and after maintenance for 21 days. Forced loss of 20% of body weight in the first week led to significantly increased serum corticosterone levels and adrenal gland weights compared to ad libitum fed controls. Serum corticosterone levels and adrenal gland weights in rats maintained at 80% of their initial body weight for 21 days remained higher than ad libitum fed control rats. However, rats maintained at 80% of an ad libitum fed control rat's weight did not differ from control rats in serum corticosterone levels or adrenal gland weights at the end of the 21-day study period. Adjustment of the feeding regimen in this manner eliminated physiological evidence of chronic stress.     

Effects of adipokinetic hormone and its related peptide on maintaining hemolymph carbohydrate and lipid levels in the two-spotted cricket,Gryllus bimaculatus

Adipokinetic hormone (AKH) regulates energy homeostasis in insects by mobilizing lipid and carbohydrate from the fat body. Here, using RNA sequencing data, we identified cDNAs encoding AKH (GbAKH) and its highly homologous hormone AKH/corazonin-related peptide (GbACP) in the corpora cardiaca of the two-spotted cricket, Gryllus bimaculatus. RT-PCR revealed that GbAKH and GbACP are predominantly expressed in the corpora cardiaca and corpora allata, respectively. Phylogenetic analysis confirmed that the identified GbAKH and GbACP belong to the clades containing other AKHs and ACPs, respectively. Injection of synthetic GbAKH and GbACP elevated hemolymph carbohydrate and lipid levels and reduced food intake significantly. In contrast, knockdown of GbAKH and GbACP by RNA interference increased the food intake, although hemolymph lipid level was not altered. Collectively, this study provides evidence that ACP regulates hemolymph carbohydrate and lipid levels in cricket, possibly collaborative contribution with AKH to the maintenance of energy homeostasis.     

The effect of food deprivation on the plasma free amino acid levels of sturgeon,Acipenser transmontanus

Synopsis Plasma levels of free amino acids were compared in recently fed sturgeon and in those deprived of food for over two weeks. Fed animals had elevated levels of branched chain amino acids. It is suggested that the higher levels of these amino acids are due to reduced metabolism of these amino acids as well as high levels in the food.     

The effects of wintertime undernutrition on plasma leptin and insulin levels in an arctic ruminant, the reindeer

We examined the effects of prolonged undernutrition on plasma leptin and insulin levels and some serum protein metabolites in reindeer (Rangifer tarandus tarandus L.) during winter and spring. The reindeer (male < 1 year) were fed their preferred winter feed, low-protein lichen ad libitum for 5 weeks, followed by 40% restriction of energy for 8 weeks and refeeding with high-protein pellets for 6 weeks. The control group received high-protein reindeer pellets ad libitum throughout the experiment. Plasma leptin decreased by 46% and insulin by 54% in the lichen group already during the ad libitum period between January and February, with parallel decreases in body weight, serum total proteins, albumin and urea. Leptin remained low during most of the energy restriction period in March and April, but increased at the end of April while body weight decreased. During the refeeding period in May and June, the body weight and insulin of the lichen group increased in parallel with total proteins and urea, but leptin remained unchanged. Similar significant reductions in plasma leptin (40%) as in the lichen group also took place in the control group fed high-protein pellets ad libitum in January and February, although their feed intake, serum total proteins and body weight remained unchanged. The results show that leptin decreases in reindeer during mid-winter, independent of food or protein intake, and suggest that the decrease may be cued by seasonal factors such as the short photoperiod.     

Effect of chronic infusion of olanzapine and clozapine on food intake and body weight gain in male and female rats

Many antipsychotics cause weight gain in humans, but usually not in rats, when injected once or twice daily. Since blood antipsychotic half-lives are short in rats, compared to humans, chronic administration by constant infusion may be necessary to see consistent weight gain in rats. Male and female rats were implanted with mini-pumps for constant infusion of olanzapine (5 mg/kg/day), clozapine (10 mg/kg/day) or vehicle for 11 days. Food intake and body weight were measured; blood drug levels were measured by HPLC. Olanzapine increased food intake and body weight in female, but not male rats. Serum olanzapine concentrations were 30-35 ng/ml. Clozapine had no effect on food intake or body weight in female or male rats. Serum clozapine concentrations were about 75 ng/ml. Single-dose pharmacokinetic analysis revealed a serum terminal half-life of 1.2-1.5 h for each drug, with no sex differences. Despite the fact that olanzapine and clozapine promote weight gain in humans, these drugs appear to have minimal effects on body weight and food intake in rats, except for a modest effect of olanzapine in female rats, even though therapeutic levels of olanzapine are achieved in serum during chronic infusion. Hence, the rapid clearance of drug following single administration in previous studies cannot explain the weak or absent effects of antipsychotics on weight gain in this species. The rat thus appears to be an inadequate model of weight gain produced by some antipsychotics in humans.     

Effects of food deprivation and high fat diet on immunoreactive dynorphin A(1–8) levels in brain regions of Zucker rats

The levels of immunoreactive dynorphin A(1–8) (ir-DYN8) were measured in discrete brain regions of lean Zucker rats subjected to food deprivation for 72 hr and to a high fat diet, and in fatty Zucker rats after food deprivation for 72 hr. Fatty rats showed higher concentrations of ir-DYN8 in the cortex and midbrain, when compared to lean rats fed a stock diet ad lib. Food deprivation increased ir-DYN8 levels in the cortex of lean rats and fatty rats and in the hippocampus of fatty rats, but decreased its content in the striatum of lean rats and in the midbrain of fatty rats. The high fat diet increased ir-DYN8 levels in the cortex and midbrain of lean rats. These results suggest that ir-DYN8 levels in extrahypothalamic structures of Zucker rats could be differentially modified under conditions of hereditary obesity and dietary manipulations.     

The effect of food and water deprivation on post-stress analgesia in mice and levels of beta-endorphin and dynorphin in blood plasma and hypothalamus

Pain sensitivity of food and/or water-deprived male mice was tested on a hotplate. The most pronounced analgesia ensued in animals given no food and water, and no food but water ad libitum, the least one in water-deprived mice. The magnitude of the rise in pain threshold depended on the duration of deprivation and was correlated with the increase in the blood plasma beta-endorphin level. In the hypothalamus beta-endorphin level increased after 72-h food deprivation only. The level of dynorphin remained unchanged. Naloxone (10 mg/kg) almost completely reversed food or water-deprivation induced analgesia.     

双歧三联活菌胶囊联合参苓白术散对 腹泻型肠易激综合征患者血浆生长抑素和神经肽Y水平影响及疗效观察

目的 探讨双歧三联活菌胶囊联合参苓白术散对腹泻型肠易激综合征(IBS)患者血浆生长抑素(SS)和神经肽Y(NPY)水平影响及疗效观察.方法 选择64例腹泻型IBS患者,随机分为观察组与对照组.两组患者均予以双歧三联活菌胶囊3粒/次,2次/d.观察组在此治疗基础上加用中药方参苓白术散,1剂/d,两组疗程均为6周.观察并比较治疗前后血浆SS和NPY水平的变化,并分析治疗后临床效果及不良反应.结果 治疗6周后,两组患者血浆SS水平较治疗前明显下降(P＜0.05和P＜0.01),NPY水平较治疗前明显上升(P＜0.05和P＜0.01),观察组下降或上升幅度明显大于对照组(P＜0.05);观察组治疗总有效率高于对照组(x2=4.27,P＜0.05),观察组患者治疗期间发生3例药物不良反应,对照组发生1例,症状均较轻,两组比较差异无统计学意义(x2=0.27,P＞0.05).结论 双歧三联活菌胶囊联合参苓白术散治疗腹泻型IBS效果良好,安全性较好,其机制可能与其降低血浆SS水平,升高血浆NPY水平密切相关.     

Effects of food deprivation on expression of growth hormone receptor and proximate composition in liver of black seabream Acanthopagrus schlegeli

The effects of food deprivation on the hepatic level growth hormone receptor (GHR) were investigated in black seabream (Acanthopagrus schlegeli) both at the protein level (by radioreceptor assay) and at the mRNA level (by ribonuclease protection assay). Serum levels of growth hormone (GH) and triiodothyronine (T₃) were also measured. Condition factor and hepatic proximate composition of the fish were also assessed. Significant decrease in hepatic GHR binding was recorded as early as on day 2 of starvation. On day 30 this decrease was even more pronounced, with the level in the starved fish reaching less than 20% the fed control level. A concomitant decrease in the hepatic GHR mRNA content was also noted during this period, with a progressive decrease from day 2 to day 30 of starvation. The extent of decrease in the mRNA content was less pronounced than the decrease in receptor binding, with the hepatic GHR mRNA content in the day 30 starved fish representing approximately 30% of the level in the fed control. In large contrast, serum GH level increased progressively during starvation. After 30 days of starvation, serum GH levels in the starved fish were more than three times the concentration found in the fed control. Serum T₃ levels, on the other hand, decreased during starvation, with the difference reaching significance on day 15 and day 30. After 30 days of starvation, serum T₃ levels in the starved fish were only approximately 40% the concentration found in the fed control. The hepatic lipid content exhibited an increasing trend during starvation. On day 30 the hepatic lipid content of the starved fish had doubled the level found in the fed control. However, the hepatic protein content did not exhibit much change during starvation. There was also a minor decrease in the moisture content of the liver during starvation, but the condition factor of the fish as a whole registered a gradual decrease during the course of food deprivation.     

The effect of dietary copper on rat plasma apolipoprotein B,E plasma levels,and apolipoprotein gene expression in liver and intestine

The plasma levels of apo B and apo E, and the level of hepatic and intestinal mRNA coding for these apolipoproteins were investigated in weanling male rats pair-fed for 6 wk with a control or copperdeficient diet. Plasma cholesterol, triglycerides, and phospholipids were significantly increased, and plasma apo B and apo E levels were also markedly increased in copper-deficient rats as compared to control rats. Copper deficiency significantly increased triglyceride levels and decreased cholesterol levels in the liver. No major differences in the levels of hepatic and intestinal apo B and apo E mRNA occurred between control and copper-deficient rats. These data imply that hypertriglyceridemia dn hypercholesterolemia owing to the copper deficiency are not accompanied by modifications in the gene expression at the mRNA level in the liver and intestine of the apolipoproteins studied.     

Effects of low molecular weight soybean peptide on mRNA and protein expression levels of differentiation markers in normal human epidermal keratinocytes

Low molecular weight soybean peptide (LSP) was applied to normal human epidermal keratinocytes, and the results showed a significant increase in the gene expression levels of involucrin, transglutaminase, and profilaggrin. Filaggrin protein levels were also significantly higher. It is possible that LSP has an epidermal cell differentiation-promoting effect and may be able to regulate metabolism of the epidermis.     

The impact of acute lung injury,ECMO and transfusion on oxidative stress and plasma selenium levels in an ovine model

The purpose of this study was to determine the effects of smoke induced acute lung injury (S-ALI), extracorporeal membrane oxygenation (ECMO) and transfusion on oxidative stress and plasma selenium levels. Forty ewes were divided into (i) healthy control (n = 4), (ii) S-ALI control (n = 7), (iii) ECMO control (n = 7), (iv) S-ALI + ECMO (n = 8) and (v) S-ALI + ECMO + packed red blood cell (PRBC) transfusion (n = 14). Plasma thiobarbituric acid reactive substances (TBARS), selenium and glutathione peroxidase (GPx) activity were analysed at baseline, after smoke injury (or sham) and 0.25, 1, 2, 6, 7, 12 and 24 h after initiation of ECMO. Peak TBARS levels were similar across all groups. Plasma selenium decreased by 54% in S-ALI sheep (1.36 ± 0.20 to 0.63 ± 0.27 μmol/L, p < 0.0001), and 72% in sheep with S-ALI + ECMO at 24 h (1.36 ± 0.20 to 0.38 ± 0.19, p < 0.0001). PRBC transfusion had no effect on TBARS, selenium levels or glutathione peroxidase activity in plasma. While ECMO independently increased TBARS in healthy sheep to levels which were similar to the S-ALI control, the addition of ECMO after S-ALI caused a negligible increase in TBARS. This suggests that the initial lung injury was the predominant feature in the TBARS response. In contrast, the addition of ECMO in S-ALI sheep exacerbated reductions in plasma selenium beyond that of S-ALI or ECMO alone. Clinical studies are needed to confirm the extent and duration of selenium loss associated with ECMO.     

长期异相睡眠剥夺对大鼠能量代谢及血清甲状腺素水平的影响

目的：探讨长期异相睡眠剥夺对大鼠能量代谢及血清甲状腺素水平的影响。方法：采用小平台水环境法建立长期异相睡眠剥夺大鼠模型;检测其能量代谢变化;放射免疫法检测血清中甲状腺素水平。结果：睡眠剥夺后大鼠摄食量由（75.06±25.37）g/（d.kg）增加到（122.30±20.43）g/（d.kg）,体重由（360.89±43.01）g减轻到（295.97±37.95）g,体温由（37.62±1.12）℃先升高到（39.00±0.87）℃后又降低至（37.72±0.84）℃,基础代谢率由（1.69±0.36）mlO2/（g.h）增加到（2.40±0.09）mlO2/（g.h）与对照组相比差异显著（P〈0.05）;血清中游离三碘甲状腺原氨酸（FT3）水平由（3.38±0.88）pmol/L降低到（2.38±0.83）pmol/L,游离甲状腺素（FT4）由（14.62±3.62）pmol/L降低到（8.26±2.80）pmol/L与对照组相比差异显著（P〈0.05）。结论：长期异相睡眠剥夺可以显著影响大鼠的能量代谢和血清甲状腺素水平。     

The influence of gradual water acidification on the acid/base status and plasma hormone levels in carp

Carp ( Cyprinus carpio ) fitted with arterial catheters were subjected to gradual water acidification (from pH 7.6 to 4.0 in 4 h), and then monitored for 48 h. The measured blood parameters showed little or no disturbance; there was no ionoregulatory disturbance, no hyperglycemia, increase in plasma lactate, hypoxemia, or swelling of red blood cells. Only a slight transient decrease of plasma pH was found. Plasma catecholamine levels remained at control level. The plasma cortisol concentration showed a transient increase, and was returned to the control level 24 h after beginning water acidification.
Our results contrast with literature data on carp where exposure to pH 4.0 caused severe blood electrolyte disturbance. We conclude that the rate of water acidification (gradual in our study as compared to acute in the literature) rather than the pH level itself determines the early effects of water acidification.     

Sex differences in the effects of chronic stress and food restriction on body weight gain and brain expression of CRF and relaxin‐3 in rats

This study investigated sex‐specific effects of repeated stress and food restriction on food intake, body weight, corticosterone plasma levels and expression of corticotropin‐releasing factor (CRF) in the hypothalamus and relaxin‐3 in the nucleus incertus (NI). The CRF and relaxin‐3 expression is affected by stress, and these neuropeptides produce opposite effects on feeding (anorexigenic and orexigenic, respectively), but sex‐specific regulation of CRF and relaxin‐3 by chronic stress is not fully understood. Male and female rats were fed ad libitum chow (AC) or ad libitum chow and intermittent palatable liquid Ensure without food restriction (ACE), or combined with repeated food restriction (60% chow, 2 days per week; RCE). Half of the rats were submitted to 1‐h restraint stress once a week. In total, seven weekly cycles were applied. The body weight of the RCE stressed male rats significantly decreased, whereas the body weight of the RCE stressed female rats significantly increased compared with the respective control groups. The stressed female RCE rats considerably overate chow during recovery from stress and food restriction. The RCE female rats showed elevated plasma corticosterone levels and low expression of CRF mRNA in the paraventricular hypothalamic nucleus but not in the medial preoptic area. The NI expression of relaxin‐3 mRNA was significantly higher in the stressed RCE female rats compared with other groups. An increase in the expression of orexigenic relaxin‐3 and misbalanced hypothalamic‐pituitary‐adrenal axis activity may contribute to the overeating and increased body weight seen in chronically stressed and repeatedly food‐restricted female rats .