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AIM: To investigate the effects of leptin on the expression of bile salt export pump (BSEP) and signaling pathway in human hepatocellular carcinoma cell line HepG2. METHODS: HepG2 cells were cultured in vitro. Leptin at concentrations of 10-8, 10-7 and 10-6 mol/L was used as a stimulating factor. The protein levels of adenosine monophosphate-activated protein kinase alpha subunit (AMPKa), phosphorylated AMPKa (p-AMPKa) and BSEP in the HepG2 cells at 24 h, 48 h and 72 h were detected by Western blotting. The optimal culture time and leptin concentration were selected, and compound C at concentration of 10 μmol/L was added to this group. The protein expression of BSEP was detected by Western blotting. RESULTS: Intervention of HepG2 cells with leptin for 72 h increased the protein expression of AMPKa gradually in a concentration-dependent manner, and leptin at concentration of 10-6 mol/L induced the strongest AMPKa expression (P<0.01). Intervention of HepG2 cells with leptin for 24 h increased the phosphorylation level of AMPKa gradually in a dose-dependent manner (P<0.01). The effect of leptin on the increase in the protein expression of p-AMPKa was also in a time-dependent manner (P<0.01). After intervention with different concentrations of leptin for 24 h, the protein expression of BSEP in the HepG2 cells was gradually increased by the stimulation of leptin in a concentration- and time-dependent manner (P<0.01). Compared with NC group, the protein expression of BSEP in 10-6 mol/L leptin group and 10-6 mol/L leptin+10 μmol/L compound C group was increased at 72 h (P<0.01), and that in 10-6 mol/L leptin+10 μmol/L compound C group was lower than that in 10-6 mol/L leptin group at 72 h (P<0.01). CONCLUSION: Leptin promotes the protein expression of BSEP in HepG2 cells by leptin-AMPK-BSEP signaling pathway. Leptin promotes the increases in AMPKa protein and the level of phosphorylation of AMPKa in HepG2 cells. 相似文献
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将鸡Leptin成熟肽的cDNA片段插入表达载体pRSET A的Nhe Ⅰ和Hind Ⅲ两位点之间,构建重组表达质粒pLep -SCAU2并转化大肠杆菌(Escherichia coli )菌株BL21(DE3), 将重组菌在含氨苄青霉素100 μg/mL 的LB培养基中培养至OD600大于0.6之后,经IPTG 0.05 mol/L诱导表达出分子量约为17.5 kD的含6×His标记的重组鸡Leptin,诱导4 h后表达量达最高,占总菌体蛋白的25%左右,且以包涵体形式存在。该重组鸡Leptin经50%Ni-NTA树脂纯化和透析复性折叠后分离出可溶性部分。对35日龄的矮脚黄鸡腹腔注射该可溶性重组鸡Leptin(2 mg/kg 体重);注射后60 min内的采食量与对照组差异不显著 (P > 0.05),但在注射后80~120 min都显著低于对照组(P < 0.05)。Leptin处理公鸡的采食量在注射2 h后逐渐上升并在5.5 h时与对照组公鸡相同。但Leptin处理母鸡的采食量持续受到抑制,在注射后5.5 h时仍然显著低于对照组母鸡(P <0.01)。表明所表达的重组鸡Leptin能显著抑制鸡的采食量(母鸡比公鸡效果更为明显),证明所制备的重组鸡Leptin融合蛋白具有生物学活性。 相似文献
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本试验研究不同饲喂次数对育肥猪生长性能的影响。采用单因子实验设计,将24头长大二元生长肥育母猪(平均初始体重60kg),分为3个处理,每个处理4个重复,每个重复2头猪。3个处理分别为:(1)分别在7∶00和15∶30各投料一次;(2)分别在7∶00、11∶30和17∶00投料;(3)在7∶00、11∶30、15∶30和20∶00投料。每次投料后,各栏猪几乎不再采食时分别将剩余饲料收回。试验期为2个月,期间以周为单位记录增重和采食量,在试验开始、中期和结束时各进行一次背膘厚度测定和血样采集。结果表明:(1)饲喂4次的生长肥育猪在采食量和日增重上均高于饲喂3次和2次的处理组,但差异不显著,这种增重作用在第1、2周较为明显(P<0.05);但料重比又均低于其他两处理组;(2)在背膘厚度上随着饲喂顿次的增加有逐渐上升的趋势,Leptin浓度随着饲喂顿次的增加也呈上升趋势,日饲喂4次比日饲喂2次和3次分别提高21.71%(P<0.05)、27.51%(P<0.05),背膘厚度与Leptin浓度呈正相关。综合以上结果,本试验认为,生长肥育母猪(平均初始体重60kg)日饲喂2次较为合理。 相似文献
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Leptin in ruminants. Gene expression in adipose tissue and mammary gland, and regulation of plasma concentration 总被引:13,自引:0,他引:13
Chilliard Y Bonnet M Delavaud C Faulconnier Y Leroux C Djiane J Bocquier F 《Domestic animal endocrinology》2001,21(4):271-295
This paper reviews data on leptin gene expression in adipose tissue (AT) and mammary gland of adult ruminants, as well as on plasma leptin variations, according to genetic, physiological, nutritional and environmental factors. AT leptin mRNA level was higher in sheep and goat subcutaneous than visceral tissues, and the opposite was observed in cattle; it was higher in fat than in lean selection line in sheep; it was decreased by undernutrition and increased by refeeding in cattle and sheep, and not changed by adding soybeans to the diet of lactating goats; it was increased by injection of NPY to sheep, and by GH treatment of growing sheep and cattle. Insulin and glucocorticoids in vitro increased AT leptin mRNA in cattle, and leptin production in sheep. Long daylength increased AT lipogenic activities and leptin mRNA, as well as plasma leptin in sheep. Mammary tissue leptin mRNA level was high during early pregnancy and was lower but still expressed during late pregnancy and lactation in sheep. Leptin was present in sheep mammary adipocytes, epithelial and myoepithelial cells during early pregnancy, late pregnancy and lactation, respectively. Plasma leptin in cattle and sheep was first studied thanks to a commercial “multi-species” kit. It was positively related to body fatness and energy balance or feeding level, and decreased by β-agonist injection. The recent development of specific RIA for ruminant leptin enabled more quantitative study of changes in plasma leptin concentration, which were explained for 35–50% by body fatness and for 15–20% by feeding level. The response of plasma leptin to meal intake was related positively to glycemia, and negatively to plasma 3-hydroxybutyrate. The putative physiological roles of changes in leptin gene expression are discussed in relation with published data on leptin receptors in several body tissues, and on in vivo or in vitro effects of leptin treatment. 相似文献
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Aykut Gokturk Uner Nesrin Sulu Cigdem Altinsaat Ahmet Ergun 《Journal of Equine Veterinary Science》2013
Day length-related alterations of several metabolic factors (glucose, leptin, insulin, and insulin-like growth factor-1 [IGF-1]), cytokines (interleukin-2 [IL-2], IL-4, IL-6, tumor necrosis factor-alpha [TNF-α], interferon-gamma [IFN-γ], and lymphocyte subpopulations [CD2, CD3, CD4, CD8, CD19, natural killer (NK) cells) were evaluated in Arabian and Thoroughbred horses. Plasma glucose, leptin, IGF-1, insulin, and cytokines levels were measured on the longest day of the breeding season and on the shortest day of the nonbreeding season. Determination of lymphocyte subpopulations was performed by flow cytometry. Glucose and IL-2 levels, CD4:CD8 ratio, and NK cells showed variations that depended on the day length. Mean concentrations of plasma leptin were higher in Arabian horses than in Thoroughbred horses, whereas mean concentrations of IGF-1 and IL-2 were lower in Arabian horses. Day length-by-breed-by-gender interaction was found for insulin, IFN-γ, and IL-4 levels. An interaction was also found between day length and gender for the expressions of CD2, CD3, CD8, and CD19. Correlations were detected between expression of CD8+ cells and levels of TNF-α and IFN-γ and between percentages of NK cells and levels of IGF-1, insulin, and glucose. Results suggested that day length and, therefore, season are important determinants or factors in modulating the immune system and could affect lymphocyte subpopulations depending on the sex of the horse. Additionally, it seems that a complex relationship in horses, as in humans and mice, exists between the immune and metabolic system, which changes according to day length, breed, and gender. 相似文献
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Leptin is thought to be not only a satiety factor but also a stimulator of angiogenesis. We examined leptin, PPARγ2, and vascular endothelial growth factor (VEGF) expression in bovine intramuscular preadipocyte (BIP) cells during proliferation. The cells were seeded at 0.85 × 104 cells/cm2 and collected every day until the fifth day after passage. Leptin mRNA was present in the cells between days 2 and 4, as indicated by RT-PCR analysis. Western blot analysis showed a band for leptin at approximately 16 kDa on all of the days during growth, and the cytoplasmic concentration of leptin was highest on day 2 and decreased gradually thereafter. A PPARγ2 band at approximately 54 kDa was also observed on all days. The concentration was highest on day 2 and decreased thereafter, which is similar to the expression pattern of leptin. In constant, the expression level of VEGF protein did not change while in culture. We have demonstrated that BIP cells can synthesize both leptin and PPARγ2, with maximal synthesis occurring during maximal proliferation. Given the role of leptin in angiogenesis, we speculate that leptin is involved in the neovascularization of adipose tissue, because new organization of adipose tissue requires the growth of new blood vessels. 相似文献
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R.S. Pleasant J.K. Suagee C.D. Thatcher F. Elvinger R.J. Geor 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2013,27(3):576-582
Background
Increased blood insulin levels are associated with an increased risk of pasture‐associated laminitis in equids.Objective
To determine the relationship between plasma insulin, leptin, and lipid levels, and measures of oxidative stress with adiposity in mature light breed horses.Animals
300 randomly selected light breed horses, aged 4–20 years.Methods
A random sample of horses (140 mares, 151 geldings, and 9 stallions) was drawn from the VMRCVM Equine Field Service practice client list. Evaluations occurred June 15 – August 15, 2006, with all sampling performed between 0600 and 1200 hours. Concentrate feed was withheld for at least 10 hours before sampling. Plasma was analyzed for insulin, glucose, leptin, triglycerides, nonesterified fatty acids, and measures of oxidative stress. Body condition score was determined as the average of 2 independent investigators.Results
Overconditioned and obese horses had higher plasma insulin (P < .001) and leptin (P < .01) levels than optimally conditioned horses. Obese horses had higher triglyceride levels (P = .006) and lower red blood cell gluthathione peroxidase activities (P = .001) than optimally conditioned horses.Conclusions and Clinical Importance
Maintaining horses at a BCS <7 might be important for decreasing the risk of pasture‐associated laminitis. 相似文献40.