Normal Feeding Group Research Articles

Changes in coagulation of sepsis rats with protein-malnutrition or energy-malnutrition

To investigate the changes in coagulation of sepsis rats with protein-malnutrition or energy-malnutrition. 108 male Sprague-Dawley (SD) rats were divided into three groups by random number table, with 36 rats in each group. The rats in normal feeding group were given a free diet (27 g/d, containing 18% protein fodder), and the rats in protein-malnutrition group were given a low protein diet (27 g/d, containing 5% protein fodder). The rats in energy-malnutrition group were given a low energy diet (9 g/d, containing 18% protein fodder). After 4 weeks of continuous feeding, 8 rats from each group were sacrificed for malnutrition evaluation. The weights of body, thymus and spleen were measured. The percentages of spleen T lymphocyte subsets and M1 macrophage were determined by flow cytometry. Plasma interleukins (IL-6 and IL-10) levels were determined by enzyme-linked immunosorbent assay (ELISA). The remaining 28 rats in each group were collected for cecal ligation and puncture (CLP) to reproduce the sepsis model, 20 rats of which were used for Kaplan-Meier survival analysis, and the other 8 rats were sacrificed at 8 hours after CLP. The levels of plasma IL-6 and IL-10 were determined by ELISA, and the percentage of spleen M1 macrophages was determined by flow cytometry. The mRNA expressions of tissue factor (TF) and plasminogen activation inhibitor-1 (PAI-1) in liver tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR). Pearson correlation method was used to analyze the correlation between the mRNA expressions of TF and PAI-1 and IL-6 in rats after CLP. (1) After 4 weeks of feeding, the rats in the normal feeding group and protein-malnutrition group gained weight, while those in the energy-malnutrition group lost 25% of their initial body weight. The weights of body, thymus and spleen in the protein-malnutrition group and the energy-malnutrition group were significantly lower than those in the normal feeding group. Compared with the normal feeding group and the protein-malnutrition group, the percentages of spleen CD3+T lymphocytes, CD4+T lymphocytes, M1 macrophages and plasma IL-6 levels were significantly increased in the energy-malnutrition group [CD3+T lymphocytes percentage: (52.1±3.7)% vs. (46.9±3.9)%, (44.5±2.2)%; CD4+T lymphocyte percentages: (35.0±3.6)% vs. (26.3±2.2)%, (25.1±2.3)%; M1 macrophage percentages: (8.7±2.0)% vs. (3.2±1.3)%, (4.2±1.1)%; IL-6 (ng/L): 129.4±16.2 vs. 48.1±10.0, 53.0±8.3, all P < 0.05]. (2) Kaplan-Meier survival analysis at 7 days after CLP showed: all rats in the energy-malnutrition group died, and the 7-day cumulative survival rate was significantly lower than that in the normal feeding group and the protein-malnutrition group [0% (0/20) vs. 35% (7/20), 55% (11/20), both P < 0.05]. The mortality of the normal feeding group was 65%, which was consistent with moderate CLP mortality, indicating that the CLP model was successfully prepared. After CLP, the plasma IL-6 level in the protein-malnutrition group was significantly lower than that in the normal feeding group [IL-6 (ng/L): 154.6±34.7 vs. 233.4±41.2, P < 0.05]. Compared with the normal feeding group, the mRNA expressions of TF and PAI-1 in liver and plasma IL-6 levels in the energy-malnutrition group were significantly increased [TF mRNA (2-ΔΔCT): 4.5±2.2 vs. 1.1±0.7, PAI-1 mRNA (2-ΔΔCT): 3.3±1.8 vs. 1.3±0.9, IL-6 (ng/L): 382.7±118.2 vs. 233.4±41.2, all P < 0.05], the percentage of M1 macrophages in spleen was significantly lowered [(8.9±2.4)% vs. (15.2±5.4)%, P < 0.05]. There was no significant difference in plasma IL-10 level among all the groups. Correlation analysis showed that the mRNA expressions of TF and PAI-1 in the liver of rats after CLP were positively correlated with plasma IL-6 level (r1 = 0.644, r2 = 0.574, both P < 0.01). Long-term sustained stress (starvation) leads to sustained chronic inflammatory state, and stimulated the release of related inflammatory factors and activation of the coagulation system after infection. And the inflammatory factors in sepsis rats without sustained stress protein malnutrition were significantly reduced.

Chewing causes rapid changes in immunoreactive nerve patterns in rat molar teeth: Implications for dental proprioception and pain

ObjectiveThis study tests the hypothesis that normal use of teeth (chewing) causes changes in immunoreactive-(IR) patterns for endings of large Aβ and CGRP axons in rat molar cusps. DesignFirst, a new paradigm to test chewing in adult male rats was developed. Then IR patterns for large dental axons were analysed for a calcium-binding protein, parvalbumin (PV), heavy neurofilament protein-200 (NFP), and vesicle-release molecule synaptophysin (SYN) that all typify large dental axons and proprioceptors for comparison with endings of CGRP-IR neuropeptide axons. The behavior groups were: (1) daytime sleeping/fasting (Group:SF); (2) brief feeding after 8–11 h of daytime sleeping/fasting (Group:SF–C); (3) normal nocturnal feeding (Group:N); (4) nocturnal fasting (Group:NF); (5) brief feeding/chewing after nocturnal fasting (Group:NF–C). ResultsNerve endings with NFP-, PV-, or SYN-IR were lost or altered in pulp and dentin in all chewing groups. Other endings with CGRP-IR were near those with PV-, NFP- and SYN-IR at the pulp-dentin border and in dentin, and they also lost immunoreactivity in all chewing groups. The special beaded regions along the crown pulp/dentin borders lost neural labeling in all chewing groups. Nerves of molar roots and periodontal ligament were not changed. ConclusionsRapid neural reactions to chewing show extensive, reversible, non-nociceptive depletions of crown innervation. Those changes were rapid enough to occur during normal feeding followed by recovery during rest. The new dental paradigm related to chewing and fasting allows dissection of intradental proprioceptive-like mechanisms during normal tooth functions for comparison with nociceptive and mechanosensitive reactions after injury or inflammation.

Long-term impact of postnatal nutritional programming on cardiac sensitivity to ischemia-reperfusion injury in vivo and on cardio-protective pathways in mice

Nutritional disturbances during the postnatal period may be responsible for a predisposition, or “programming”, to increased cardio-metabolic risk and to a progressive alteration of left ventricular contractility in adulthood. Our aim was to evaluate in mice the impact of postnatal overfeeding (PNOF) on cardiac sensitivity to ischemia-reperfusion (I-R) injury in vivo and on the myocardial expression of genes involved in heart protection against ischemia. PNOF was induced by the reduction of litter size of C57/BL6 mice immediately after birth: normally-fed group (NF) was composed of 9 male pups/mother and overfed group (OF) of 3 pups/mother. In vivo I-R injury was induced by the ligation of the anterior interventricular artery for 45 minutes followed by 24h of reperfusion in hearts from 6-months aged mice. The gene expressions of proteins of interest composing the cardioprotective pathways (RISK pathway: Akt, ERK; SAFE pathway: JAK/STAT3; Sirtuins: SIRT1) were measured by RT-qPCR in mice aged 4 and 6 months. PNOF induced in OF group an early and permanent increase in body weight (+ 23%, P < 0.01, N = 11) and a significant increase of infarct size (+ 32%, P < 0.05, N = 11) at 6 months. Hearts OF mice aged 4 months showed a decreased expression of cardioprotective pathways, represented by a reduction in the gene expression of ERK1, Akt, PI3K, STAT3 and SIRT1. At 6 months, the expression of cardioprotective pathways was also reduced in OF group, with the diminution of the gene expression of STAT3, Akt and Erk1. Nutritional programming through short-term PNOF induced a long-lasting decrease in the expression of signaling pathways involved in cardiac cellular protection, which could explain why 6-months old mice show increased susceptibility to myocardial I-R injury in vivo. The mechanism of these alterations needs further exploration, but could involve alterations in cardiomyocyte's balance between apoptotic and regenerative pathways.

Methotrexate-induced neural tube defects mouse model under folate deficient condition

Objective To establish neural tube defect mouse model induced by folate antagonist methotrexate (MTX) under low folate diet, and to demonstrate folate deficiency acting as a sensitizing agent leading to neural tube defects. Methods C57BL/6J female mice were divided into 3 groups. Normal feeding group was fed with normal SPF diet. Low folate diet group was given special low folate food. Low folate and MTX group was injected intraperitoneally with MTX (1.00 mg/kg, 1.25 mg/kg, 1.50 mg/kg, 2.50 mg/kg, 3.50 mg/kg, 4.50 mg/kg, respectively) at E7.5 and dissected at E10.5 to E13.5. The other 2 groups were injected with equal volume of normal saline. The embryo phenotype was observed and the number of normal and deformed fetuses were counted. Fetal brain to hip lengths were measured. Embryos and placentae were weighted. Results In low folate diet group, developmental retardation was observed without mouse embryonic abnormality of neural tube defects (NTDs). The MTX dose of 1.50 mg/kg might led to the highest deformity rate of 36.5% in low folate diet plus MTX group and the phenotype of NTDs was mainly spina bifida. Conclusion Methotrexate with low folate diet could be used to establish the mouse model of NTDs. Key words: Neural tube defects; Methotrexate; Models, animal; Folate antagonist

PO-212 Effects of hypoxic endurance training on weight loss and Irisin expression in nutritional obese rats

Objective Exercise can induce the release of various myokine such as Irisin, which promote browning of white fat, improve body metabolism, and loss weight. Appropriate hypoxic training plays a better role in weight loss than single exercise, but the effect of hypoxic training on muscle factors that promote browning of fat is rarely reported. Therefore, this study aims to observe different concentrations of hypoxia training on the PGC1α-Irisin-UCP1 signaling pathway in nutritional obese rats, and the role of weight loss.&#x0D; Methods Male Sprague-Dawley rats (n=140) of 6 weeks old were divided into normal diet feeding group (group N, n=20) and high-fat diet feeding group (group HFD, n=120). The HFD rats became obese after 8 weeks’ feeding, and they were further divided randomly into 8 groups. Including sedentary group(group A), training group (group AE), 16.3% hypoxia exposure group (group B), 16.3% hypoxia training group (group BE), and 13.3% hypoxia exposure group (group C), 13.3% hypoxia training group (group CE), 11.3% hypoxia exposure group (group D), 11.3% hypoxia training group (group DE), group B, group C and group D Rats were exposed to oxygen in concentrations of 16.3%, 13.3%, and 11.3% for 12 h/d, respectively. Rats in group BE, CE, and DE were subjected to animal treadmill training during hypoxic exposure with a slope of 0°, 20 m /min, 40min/d, 5d/w. After 8 weeks of intervention, blood, adipose tissue and skeletal muscle were collected and tested.&#x0D; Results (1) In group AE, the body weight of obese rats decreased in a short time, but bounced back later. The body weight of rats in group CE and group DE decreased continuously. Hypoxia exposure and hypoxia training can inhibit the food intake of obese rats in varying degrees, and group CE has the most obvious effect. Hypoxia, endurance training and hypoxia training all reduced the percentage of visceral fat to body weight, group C is significantly lower than group B and D group (p &lt; 0.05), the effect of hypoxia training is more obvious than single hypoxia or exercise, and the lower oxygen concentration the more significant effect. Group C、group CE、group D and group DE significantly decreased the concentration of LDL-C, increased the content of HDL-C in serum than other groups (p &lt; 0.05).(2) The concentration of serum Irisin in group AE, group BE, group CE and group DE rats was significantly higher than that in group A (p &lt; 0.05), and the highest level was found in group CE and DE. The insulin resistance of obese rats in group CE and group DE was significantly better than that in group AE (p &lt; 0.05). (3) Three different concentrations of hypoxia exposure and hypoxia training can significantly increase the skeletal muscle PGC-1 α and FNDC5 gene expression, showing that the lower the concentration of oxygen, the higher the expression, 13.3% and 16.3% hypoxia training can significantly promote the gene transcription of UCP1. The contents of PGC-1 α and FNDC5 protein in skeletal muscle of obese rats in group C, AE and DE were significantly higher than those in group A (p &lt; 0. 05). The expression of UCP1 protein in skeletal muscle of rats in group C, CE and DE was significantly higher than that in group A (p &lt; 0. 05).&#x0D; Conclusions 13.3% and 11.3% oxygen concentration combined with endurance training can effectively reduce the body weight and visceral fat of nutritional obese rats, enhance serum HDL-C and decrease LDL-C levels and insulin resistance. 13.3% and 11.3% oxygen concentration training can significantly improve the level of serum Irisin, 11.3% oxygen concentration training can significantly promote the expression of PGC1α-Irisin-UCP1 gene and protein in skeletal muscle.

Dewaxed Brown Rice Feed Improves Fatty Liver in Obese and Diabetic Model Mice.

Dewaxed brown rice has macrophage activation ability via TLR4 and contains a high amount of lipopolysaccharides (LPS). It is expected that dewaxed brown rice can help prevent lifestyle diseases. In this study, the anti-obesity effect of dewaxed brown rice was investigated using obese and diabetic model mice. Dewaxed brown rice and white rice were polished and powdered by Toyo Rice Co. Diet pellets were prepared (AIN-93) with 50% dewaxed brown rice or white rice powder and fed to type II diabetic model KK-Ay mice for 10 weeks. Weight and fasting blood glucose were measured every week, and whole blood and liver was collected on the final day for the evaluation of biochemical data. A 20% reduction in body weight was found in the dewaxed brown rice feed and white rice feed groups compared to the normal feed group. Fasting blood glucose increased in the normal-diet group, but on the other hand, the blood glucose in the white rice and the dewaxed brown rice feed group was almost constant. Dewaxed brown rice feed group of plasma ALT and AST, liver TG and T-CHO were significantly lower than that of the control and the white rice feed group. Dewaxed brown rice feed has an anti-obesity effect to suppress increasing body weight, fasting blood glucose, and an effect of suppressing fatty liver.

Mechanism Mediated by S6 Kinase Resulting in both Sufficient Glucose Metabolism Improvement and Insufficient Weight Loss by Glucose Excretion against Fat Load

SGLT2 inhibitors have succeeded in improving prognosis in addition to blood glucose control and weight loss in T2D patients, but the reason is unclear, Systemic metabolic changes when improvement by glucose excretion is not sufficiently elucidated. Here, metabolic respiration balance and insulin signal change were analyzed when the high fat diet-induced insulin resistance was improved with the SGLT2 inhibitor tofogliflozin. 8-week-old male C57/Bl6 mice were divided into 3 groups; normal feed group (N group), 60% high fat (HF) feed group and 3 mg/kg/day tofogliflozin (Tofo) administered group on high fat diet. After 8 weeks of food loading, the body weight increased by 30% in the HF group compared to the N group and by 16% in the Tofo. Both IPGTT and IPITT were improved in the Tofo compared with HF group. In the calorimetric analysis, the heat production increased, the respiratory exchange ratio (RER) decreased, and the RER diurnal variation disappeared in the HF group, while the Tofo reduced the energy consumption to the same level as the N group while maintaining the RER equivalent to the HF group. UCP1 in brown fat was elevated in the HF group and improved in the Tofo. In liver, muscle and fat IRS-1 amount decreased in the HF group and revived in Tofo. Phosphorylation of the basal S6K was enhanced in the HF group and disappeared in the Tofo, while after insulin administration, pAkt and resulting pS6K were accelerated in the Tofo. In conclusion, urinary glucose excretion against high fat load is resulting in improvement of IRS-1 degradation by basal S6Kinase promotes insulin Akt activation. Lipid synthesis is enhanced by insulin while energy expenditure is normalized. Inhibited fat burning compensation leaves weight gain, but since insulin signal has been improved enough, glucose metabolism is improved adequately. This suppression of catabolism might be involved in the improved prognosis by administration of SGLT2 inhibitors. Disclosure T. Kikuchi: None. A. Kushiyama: Research Support; Self; Teijin Pharma Limited, Sanwa Kagaku Kenkyusho Co., Ltd., Kowa Pharmaceutical Co., Ltd, StaGen. Consultant; Self; RIZAP. M. Fujishiro: Research Support; Self; Johnson &amp; Johnson Services, Inc.. H. Sakoda: None. T. Asano: Research Support; Self; Teijin Pharma Limited, Sanwa Kagaku Kenkyusho Co., Ltd., Sanofi K.K. Y. Iwamoto: Board Member; Self; Japan Diabetes Foundation.

Effects of bone marrow mesenchymal stem cells on blood glucose concentration in diabetic rats

Objective To investigate the effects of different concentrations of bone marrow mesenchymal stem cells (BMSCs) on blood glucose concentration in diabetic rats. Methods SD rats’ bone marrow was isolated, purified and cultured to obtain BMSCs. The expression of surface markers CD34, CD44, CD45 and CD90 was detected. The models of diabetic rats were randomly divided into control group, the low concentration group, high concentration group and normal feeding group. Low concentration group and high concentration group were injected with 1×105 and 5×106 BMSCs respectively, and the normal group and the control group were injected with equal volume of phosphate buffer (PBS). The dynamic changes of blood glucose concentrations at 1st, 2nd and 3rd week after injection were statistically analyzed. Results BMSCs were proved by immunofluorescence with CD34 (-), CD44 (+ ), CD45 (-) and CD90 (+ ). After treatment with BMSCs, blood glucose had no significant change in the first two weeks, but decreased by 16.1% at 3rd week in the low concentration group. As compared with the control group, blood glucose concentration was significantly decreased, and decreased by 42.0% at 3rd week in the high concentration group. There was no obvious change in blood glucose concentration before and after treatment in the normal and control groups. Conclusion High concentration (5×106 cells) BMSCs can control and regulate the blood glucose concentration in diabetic rats. Key words: Bone marrow mesenchymal stem cells; Diabets; Rat; Blood glucose concentration

Long-term impact of postnatal overfeeding on myocardial protective pathways

Nutritional disturbances during the postnatal period may be responsible for a predisposition in adulthood to increased cardio-metabolic risk and a greater myocardial vulnerability to ischemia. This nutritional perinatal programming may also lead to an alteration of cellular pathways involved in cardiac protection, such as the specific RISK and SAFE pathways, highlighted during pre- and post-ischemic conditioning or those of sirtuins (SIRT), histone deacetylases involved in the regulation of essential biological process. Therefore, our goal was to evaluate the impact of postnatal overfeeding (PNOF) on myocardial expression of genes involved in heart protection against ischemia. PNOF was induced by reduction of litter size of C57/BL6 mice immediately after birth: normally-fed group (NF) was composed of 9 pups/mother and overfed group (OF) of 3 pups/mother. The gene expressions of proteins of interest composing the cardioprotective pathways (RISK pathway: Akt, ERK, SAFE pathway: JAK/STAT3, Sirtuins: SIRT1) were measured by RT-qPCR in mice aged 24 days, 4 and 6 month. PNOF induced an early and permanent increase in body weight in OF group. An early rise (at 24 days) of the gene expression of JAK, ERK1 and SIRT-1 was observed in OF group, whereas at 4 months the expression of these cardioprotective pathways was decreased, represented by a reduction in the gene expression of ERK1, Akt, JAK, STAT3 and SIRT1. To conclude, over time, in PNOF-treated mice, a decrease in the expression of signaling pathways involved in cardiac cellular protection occurs, which could explain why adult animals show increased susceptibility to cardiac ischemic lesions. The mechanism of this alteration of expression is not yet understood, but it could involve epigenetic modifications.

Effects of various LED light spectra on circadian rhythm during starvation in the olive flounder (Paralichthys olivaceus)

In aquaculture, feeding is essential for the maintenance of metabolic processes and homoeostasis of fish. However, fasting acts as a stressor. In this study, we investigated the effect of circadian rhythm under various LED wavelengths [blue (460 nm), green (520 nm) and red (630 nm)] and two light intensities (0.3 and 0.6 W m−2) over a 9-days period in the olive flounder (Paralichthys olivaceus). We analysed clock genes like period 2 (Per 2) and cryptochrome 1 (Cry 1), and serotonin and arylalkylamine-N-acetyltransferase 2 (AANAT 2), which control circadian rhythms. Per 2, Cry 1, serotonin and AANAT 2 were significantly decreased during the starvation period compared to the normal feeding group. Nevertheless, their levels increased in the groups exposed to green- and blue LED light during the experimental period. These results confirmed that green and blue wavelengths are effective in maintaining the circadian rhythm in olive flounder.

Effect of chronic low potassium on K+ uptake rate in myocardium and skeletal muscle of rabbits

Objective To evaluate the effect of chronic low potassium on K+ uptake rate in the myocardium and skeletal muscle of rabbits. Methods Thirty-two adult male rabbits, aged 12-14 weeks, weighing 2.0-2.7 kg, were randomly divided into 4 groups (n=8 each) using a random number table: normal feeding group (group N), low potassium feeding group (group L), potassium supplementation control group (group SC) and potassium supplementation experimental group (group SE). N and SC groups were given a normal diet only, and L and SE groups were fed with a low potassium diet for 15 days. Potassium chloride (KCl) 0.5 mol/L was then infused intravenously at the initial rate of 60 μmol·kg-1·min-1 in SE and SC groups. Blood samples were obtained from the central artery of the left ear every 5 min for measurement of plasma K+ concentrations. The infusion rat of KCl was then adjusted until the plasma K+ concentration reached 5.5 mmol/L and maintained at this level for 1 h, and then infusion was stopped. The total volume of KCl infused was recorded. The hearts and soleus muscle of animals were excised for determination of K+ content. K+ uptake and uptake rate were calculated. Results Compared with N group, the plasma K+ concentration, and K+ content in the myocardium and soleus muscle were significantly decreased in group L(P<0.05). Compared with SC group, the total volume of KCl infused, and K+ uptake and uptake rate in the myocardium and soleus muscle were significantly increased in group SE(P<0.05). Conclusion Chronic hypokalaemia can increase K+ uptake rate in the myocardium and skeletal muscle of rabbits. Key words: Hypokalemia; Myocardium; Muscle, skeletal

Effects of overfeeding in breastfeeding period on blood pressure and vascular endothelial dilation function of rats

Objective To study the effects of postnatal overfeeding and high-fat diet on blood pressure of rats, and to explore the pathophysiological mechanism underlying hypertension induced by continuous early postnatal overfeeding. Methods Male Sprague-Dawley rats were randomly divided into normal feeding group (10/litter) and overfeeding group (3/litter) on postnatal day 3 with a random number table. After weaning at postnatal week 3, the rats were randomly given standard chow or high-fat (HF) diet until week 16. Hence four groups were analyzed, namely normal feeding group, breastfed overfeeding group, post-weaning overfeeding group, and continuous overfeeding group. Body weight was continuously monitored in each week. Visceral fat pad(retroperitoneal and perigenital), systolic pressure, and heart rate were observed at week 3 and week 16. Thoracic aorta was sampled for measurement of vascular endothelial dilation function. Histological morphology was observed with HE staining, nitric oxide content of thoracic aorta was detected with nitrate reductase method. The mRNA expression of endothelial nitric oxide synthase (eNOS) in thoracic aorta was determined by real-time polymerase chain reaction. The protein expressions of eNOS and phosphorylated eNOS were determined by Western blot. Results At week 3, breastfed overfeeding rats displayed significantly larger body weight [(77.80±0.57) g vs. (62.80±0.85)g, t=14.576, P<0.01] and visceral fat [retroperitoneal: (8.19±0.49)mg/g vs. (4.92±0.31)mg/g, t=5.629, P<0.01; perigenital: (3.50±0.29)mg/g vs. (2.08±0.13)mg/g, t=4.552, P<0.01] compared with normal feedindg rats, and the protein expression of phosphorylated eNOS in aortic tissues was significantly reduced to week 16 (F=15.215, P<0.01); high-fat diet feeding after weaning further increased the body weight and fat mass in breastfed overfeeding rats. At week 16, continuous overfeeding rats showed hypertension [(149±1.94)mmHg (1 mmHg=0.133 kPa), F=22.834, P<0.01], impaired vascular endothelial dilation function (F=7.648, P<0.05), and reduced protein expression of phosphorylated eNOS(F=15.215, P<0.01), while the post-weaning overfeeding group only had elevated blood pressure. Conclusions Overfeeding in breastfeeding period and high-fat diet after weaning leads to hypertension. The continuous decrease in phosphorylated eNOS in vascular tissues may be an important molecular process participating in the occurrence of vascular endothelial dysfunction in adults induced by postnatal overfeeding. Key words: Early postnatal overfeeding; Vascular endothelial dilation function; Hypertension; Nitric oxide; Phosphorylated endothelial nitric oxide synthase

Protein Restriction Produces Alterations in Nitrergic Myenteric Neurons in the Proximal Colon in Rats

Protein Restriction Produces Alterations in Nitrergic Myenteric Neurons in the Proximal Colon in Rats Catchia Hermes-Uliana, Marcelo Biondaro Góis, LetíciaSarturi Pereira-Severi, Eduardo José de Almeida Araújo, Débora de Mello Gonçales Sant’Ana Abstract Aim: This study aimed to investigate the effects of severe protein restriction on the morphometric and quantitative aspects of neurons in the myenteric plexus of the proximal colon in rats. Methodology: Ten rats were divided into two groups: (i) a normally fed group (NG) that received commercial chow with 26% protein for 90 days and (ii) a protein restriction group (RG) that received chow that contained a reduced amount of protein (4%) for 90 days. Nitrergic neurons were evaluated by nicotinamide adenine dinucleotide phosphate (NADPH-diaphorase) histochemistry. Intestinal segments were dissected. The number of neurons was counted, and the area of cellular bodies was measured. Results: A significant (58.92%) increase in the number of neurons that expressed NADPH-diaphorase and significant decreases in the area of cellular bodies, nuclei, and cytoplasm were found in the RG compared with the NG. Conclusion: In conclusion, protein restriction (from 26% to 4%) increased neuronal population density and nitrergic myenteric neuron atrophy in the proximal colon in rats. Full Text: PDF DOI: 10.15640/ijhs.v3n1a10

Effect of biphenyl dimethyl dicarboxylate and fluoxetine on performance of normally-fed and protein malnourished psychologically stressed mice in elevated plus maze

Stress alters psychological diseases such as anxiety and depression. Protein malnutrition (PM) contributes to psychological disorders. The present study aimed to investigate the effect of biphenyl dimethyl dicarboxylate (DDB) on anxiety of psychologically stressed protein malnourished mice as compared to its effect in normally-fed mice. Fluoxetine (FLX) was used as reference standard. Animals were randomly divided into two major groups, normally-fed group provided with 20% casein diet and a protein malnourished one was provided with a diet containing 8% casein for 21 consecutive days. The given diet was continued during the experiment. Stress was induced using the learned helplessness technique (LH). Each animal was exposed for 5days to the psychological stress session either alone or in association with drug administration following completion of 21days under the selected diet regimen. DDB and fluoxetine were given in doses of 100mg/kg p.o. and 10mg/kg i.p., respectively before exposure to foot shocks daily. Anxiety was evaluated using elevated plus maze (EPM) and the whole brain wet tissue monoamine levels, namely dopamine, norepinephrine and serotonin were estimated.Protein malnutrition in the presence of stress resulted in increasing number of open arm entries and head dipping as compared to the normally-fed stressed mice. Stress in the presence or absence of protein malnutrition significantly increased rearing frequency as compared to the corresponding mice. Fluoxetine in the presence of stress significantly decreased open arm time spent and number of open arm entries as compared to protein malnourished stressed mice. Protein malnutrition increased stretched attend posture.

Model building-up and observation on the mouse carried chronic hepatitis B and nonalcoholic fatty liver disease

Establish the model of mouse with chronic hepatitis B virus (HBV) and nonalcoholic fatty liver disease (NAFLD). Take 100 HBV transgenic, BALB/c mice of 4 weeks old, with each gender half. Then pick out 70 mice in one group to feed high-fat feed and the rest to feed normal feed. At the end of week 16, random kill 10 mice of high-fat, then liver tissue and serological detection target identification model is established in this paper. After that, divide the mice into model group and comparison group with 30 mice in each group. Feed model group with high-fat feed, comparison group with normal feed and normal group with normal feed till week 72 (including previous 16 weeks). Kill 10 mice of each group at the end of week 24, 48 and 72 respectively, fully automatic biochemical instrument detection of serum ALT, AST, TC, TG, FBG, fluorescence quantitative PCR method to detect HBV-DNA, chemiluminescence detection of HBsAg, liver biopsy after HE staining to evaluate histology change, observe mice model of dynamic evolution. (1) Feed high fat feed after 16 weeks, mice's weight, serum ALT, AST, TC, TG, FBG and blood biochemical indicators increased, HBV-DNA positive, liver HE staining obviously big blister fatty degeneration of liver cells and within the lobule lymphocytes infiltration, NAFLD activity score (NAS) getting close to NASH, the model of chronic HBV carries with NAFLD mouse built successfully. (2) The TC and TG values of model group in each period were higher than that of comparison group and normal group. (3) In week 24 and 72, HBV-DNA values of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05). (4) In week 48 and 72, NAS of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05). (1) Chronic HBV carries with NAFLD mice model can be established by HBV transgenic mice fed by high fat feed. (2) NAFLD accelerates the liver disease of the mice carrying HBV to some extent.

Lycopene Inhibits Ischemia/Reperfusion-Induced Neuronal Apoptosis in Gerbil Hippocampal Tissue

Plant lycopene exhibits antioxidant activity in animal tissues. Transient cerebral ischemia/reperfusion in Mongolian gerbils resulted in delayed neuronal death in hippocampal regions. We examined the antioxidant effects of lycopene because we expected lycopene to attenuate ischemia-related neuronal damage by controlling apoptosis at the gene level. The gerbils were divided into two groups: the normal feeding (control) group that received normal market food (MF) and the lycopene group that received MF containing lycopene (5mg in 100g MF food). After 1.5-2.0months (when body weight were 60-65g), the lycopene level was 38.2±17.6ng/ml in serum and 11.9±4.0μg/g-wet weight tissue in the liver. Levels of B cell leukemia-2, an apoptosis-suppressing protein, decreased in control animal brains 1, 3, and 7days after surgery, whereas the levels increased in lycopene-treated animal brains. Moreover, cysteinyl aspartate-specific protease-3 activity increased gradually after ischemia, but was suppressed in the lycopene-treated animal brains 7days after surgery. Finally, hippocampal superoxide dismutase (SOD) activity decreased in the control group 3h after ischemia and, gradually increased thereafter, whereas it was significantly elevated in the lycopene group. Thus, orally administered lycopene is accumulated in the body, and provided protections against ischemia/reperfusion-induced brain injury by inducing an increase in SOD activity and inhibiting apoptosis.

Effects of delayed first feeding on the growth,feeding and activities of digestive enzyme of theMyxocyprinus asiaticuslarvae

To investigate the effects of different starvation periods on the growth capacity,feeding and activities of digestive enzyme of the Myxocyprinus asiaticus larvae under the condition of w ater temperature( 19 ± 0. 5) ℃,two experiments were performed. The first one was a controlled experiment with one complete starvation group( S) and one normal feeding group( C). The second w as the delayed first feeding experiment along w ith the initial feeding time groups: 0( control group),1,3,5,7,9 and 11 delayed days groups,denoted as D0,D1,D3,D5,D7,D9 and D11. The total experimental period lasted 29 days. Results show ed that:( 1) Since hatching,larvae's first feeding w as on the 7th day,and the yolk w as exhausted on the14th day,w hile PNR( the point of no return) appeared on the 27th day,w ith conspicuous pectoral angle characteristics and significant increase in mortality around the PNR.( 2) After refeeding,compared to the control group,the D1 and D3 groups show ed no significant difference in body w eight,indicating a complete compensation effect.( 3) During the 1th to the 7th day starving process,the trypsin and amylase enzyme activity decreased significantly( P 0. 05),w hile w hen prolonging the starvation time to 9 and 11 days,the lipase and trypsin enzyme activity rose significantly( P 0. 05). After refeeding,w ith starvation time lasting,to the 19th day larvae,the trypsin activity significantly increased and then gradually decreased,but indicated the opposite trend on the amylase activity,to the 29th day larvae,and both of lipase and amylase show ed a significant upw ard trend. To sum up,our results indicated that starvation can cause significant changes of M.asiaticus in the larvae grow th,feeding and digestive enzymes,along w ith different compensatory grow th mechanism after various starvation time periods. After recovering feeding,the digestive enzymes recovery strategy is not consistent for larvae are w ith different starvation time. Totally,larvae are inclined to use the small carbohydrates along w ith starvation time lasting. But the inherent mechanism needs further study.

Pengaruh Air Perasan Umbi Bawang Merah (Allium ascalonicum L.) terhadap Malondialdehid (MDA) Plasma Mencit yang diinduksi Hiperkolesterolemia

Malondialdehyde, a product of lipid peroxidation, is often used as an indicator to detect the oxidative stress damaged in the body. Lipidperoxidation is associated with progression of atherosclerosis. It is known that onion (Allium ascalonicum L.) can be used as an antioxidantand hypolipidemic agent, but whether onion can decrease hypercolesterolemia mice MDA plasma induced by yolk is still unknown. Thepurpose of this study was to investigate the effect of onion’s juice on hypercolesterolemia mice MDA plasma level. This was an experimentallaboratory research with post test only design with control. The measured parameters were cholesterol and MDA level on tested groups.Groups with treatment consist of yolks group, and 80% shallot juice, simvastatin and aquadest were separately used as positive and negativecontrol. The result showed that there was significant difference between high fat feed group which added 80% onion’s juice and high fat feedgroup only. Addition of 80% onion’s juice in high fat feed groups decreased MDA plasma level equal to normal feed group. In conclusion, itwas proved that onion’s juice decreased mice MDA plasma level.

Ischaemic postconditioning reduces serum and tubular TNF-α expression in ischaemic-reperfused kidney in healthy rats

We studied the protective effects of postconditioning (PS) in healthy and hypercholesterolemic rats after renal ischaemia-reperfusion (IR) injury. We aimed to examine cytokine expression and apoptosis in tissue damage after revascularisation (TNF-α levels in serum and tissue). Male Wistar rats (n = 32) were divided into four groups. The animals of normal feed groups (NF) were fed with normal rat chow and the cholesterol feed groups (CF) were fed with 1.5% cholesterol containing diet for 8 weeks. Anaesthetized rats underwent a 45-min cross-clamping in both kidney pedicles. Ischaemia was followed by 120-min reperfusion with or without PS protocol (group PS vs. IR). Postconditioning was induced by four intermittent periods of ischaemia-reperfusion of 15-s duration each. Serum cholesterol, triglyceride, urea and creatinine levels were determined. Proinflammation was characterized by the measurement of serum TNF-α. Tissue injury in kidney was determined by formaline-fixed, paraffin-embedded tissue sections. Tissue TNF-α levels were determined by immunohistochemistry. Significant elevation was observed in serum TNF-α level after IR injury in normal feed groups, which was reduced by PS. In CF group neither the elevation nor the postconditioning induced reduction were as significant as in the NF groups. In normal feed group PS caused a significant reduction in tissue TNF-α level which was significantly higher in CF. Ischaemic postconditioning proved to be an effective defense against IR in NF groups, but it was ineffective in CF groups in kidney tissue.

The effects of NF-E2-related factor-2 prompter polymorphism on alcoholic liver disease with Vibrio vulnificus sepsis

To investigate the influence of genetic polymorphism in NF-E2-related factor-2 (nrf2) gene promoter locus at 336 in alcoholic liver disease (ALD) with Vibrio vulnificus (VV) sepsis. Through the simple random sampling method, C57B6 male mice were divided into normal feeding group (group A, 10 mice), alcoholic liver disease group (group B, 10 mice), normal feeding group infected with VV through intraperitoneal injection (group C, 8 mice), alcoholic liver disease group infected with VV (group D, 110 mice). Through gene sequencing method, nrf2 gene promoter 336 polymorphism in D group was analyzed and grouped into: non-mutation group (336T) (group D1, 7 mice) and mutation group (336C) (group D2, 10 mice). Through RT-PCR, Western-blotting and ELISA method, expressions of nrf2, tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), high mobility group protein 1 (HMGB(1)) gene and protein of liver were measured. The pathological changes in liver were recorded with light microscope. After infected with VV for 48 hours for A, B, C, D1, D2 group, the expression medians of nrf2 mRNA in liver were 0.115, 0.173, 0.211, 0.764, 0.352, respectively (χ(2) = 40.64, P < 0.05), the expression medians of IL-10 mRNA in liver were 0.338, 0.637, 1.002, 1.825, 1.403, respectively (χ(2) = 41.05, P < 0.05), the expression medians of TNF-α mRNA in liver were 0.140, 0.254, 0.372, 0.399, 0.699, respectively (χ(2) = 38.16, P < 0.05), the expression medians of HMGB(1) mRNA in liver were 0.230, 0.410, 0.668, 0.508, 1.021, respectively (χ(2) = 31.45, P < 0.05). After infected with VV 48 hours for mice in A, B, C, D1, D2 group, the expression medians of nrf2 protein in liver were 0.908, 1.461, 2.061, 3.982, 2.243, respectively (χ(2) = 33.72, P < 0.05), the expression medians of IL-10 protein in liver were 13.97, 22.54, 30.14, 57.98, 41.53, respectively (χ(2) = 37.31, P < 0.05), the expression medians of TNF-α protein in liver were 114.07, 142.94, 175.44, 174.60, 266.11, respectively (χ(2) = 32.29, P < 0.05), the expression medians of HMGB(1) protein in liver were 2.01, 6.05, 9.62, 6.24, 12.89, respectively (χ(2) = 36.94, P < 0.05). Compared with group A, there were large amount of fat drops, fatty changes in group B, inflammatory cell infiltration, disorder of hepatic cell in group C, and extension of hepatic duct and vein, edema of liver cells and disorder of hepatic cells in group D. The nrf2 gene promoter of T336C mutation in C57B6 mouse of ALD can significantly decrease the expression of nrf2, and intensify organ inflammation and damage when they were infected by VV.