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Special Issue: Lipid Metabolism, Adipogenesis and Fat Tissue Metabolism: Gene Regulation
Lipid metabolism is pivotal in controlling energy homeostasis [...]
The Effects of Neuropeptide B on Proliferation and Differentiation of Porcine White Preadipocytes into Mature Adipocytes
Neuropeptide B (NPB) affects energy homeostasis and metabolism by binding and activating NPBWR1 and NPBWR2 in humans and pigs. Recently, we reported that NPB promotes the adipogenesis of rat white and brown preadipocytes as well as 3T3-L1 cells. In the present study, we evaluated the effects of NPB on the proliferation and differentiation of white porcine preadipocytes into mature adipocytes. We identified the presence of NPB, NPBWR1, and NPBWR2 on the mRNA and protein levels in porcine white preadipocytes. During the differentiation process, NPB increased the mRNA expression of PPARγ, C/EBPβ, C/EBPα, PPARγ, and C/EBPβ protein production in porcine preadipocytes. Furthermore, NPB stimulated lipid accumulation in porcine preadipocytes. Moreover, NPB promoted the phosphorylation of the p38 kinase in porcine preadipocytes, but failed to induce ERK1/2 phosphorylation. NPB failed to stimulate the expression of C/EBPβ in the presence of the p38 inhibitor. Taken together, we report that NPB promotes the differentiation of porcine preadipocytes via a p38-dependent mechanism.
Isolation method and characterization of adipocytes as a tool for equine obesity research – In vitro study
Genistein stimulates the viability and prevents myofibroblastic transformation in human trabecular meshwork cells stimulated by TGF-β
The Effects of Peruvian maca (Lepidium meyenii) Root Extract on In Vitro Cultured Porcine Fibroblasts and Adipocytes
Peruvian maca (Lepidium meyenii) is a plant known for its nutritional and medicinal properties whose use as a supplement in animal diets has attracted much interest. We studied the effects of powdered maca root extract on the growth potential of in vitro cultured porcine cells prior to its use as an additive in animal nutrition. Fibroblast cell viability (MTT), cell proliferation (BrdU), and apoptosis level (TUNEL) were measured for a range of extract doses (0, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 7.0, and 10 mg/mL). Transcript levels of CCND1, MCM2, and PCNA genes as molecular markers of cell proliferation were also determined. Next, the effects of maca extract at 2 and 5 mg/mL on in vitro induced adipogenesis were evaluated over eight days of differentiation. The transcript levels of three adipocyte marker genes (CEBPA, PPARG, and FABPB4) were measured at days 0, 4, and 8 of adipose differentiation, and lipid droplet accumulation (BODIPY staining) was also noted. No cytotoxic effect was detected on fibroblast cell viability, and the inhibitory concentration (IC50) value was determined to be IC50 > 10 mg/mL. Doses of maca extract above 3 mg/mL decreased cell proliferation. The transcript level decreased in concentrations above 5 for the MCM2 and PCNA genes. For the CCND1 gene, the transcript level decreased when the greatest maca dose was used. In the in vitro adipogenesis experiment, it was found that the rate of lipid droplet formation increased on day 4 of differentiation for both doses, while decreased lipid droplet formation was observed on day 8 for 5 mg/mL of maca extract. Significant changes were seen in the mRNA level for CEBPA and PPARG on days 4 and 8, while the transcript of FABP4 increased only on day 8 at 2 mg/mL dose. It can be concluded that the addition of Peruvian maca in small doses (<3 mg/mL) has no negative effect on porcine fibroblast growth or proliferation, while 2 mg/mL of maca extract enhances adipocyte differentiation.
MOTS-c regulates pancreatic alpha and beta cell functions in vitro
AbstractThe aim of this study is to determine the influence of the mitochondrial open-reading-frame of the twelve S rRNA-c (MOTS-c) peptide on pancreatic cell physiology. Moreover, in this study, we examined the changes in MOTS-c secretion and expression under different conditions. Our experiments were conducted using laboratory cell line cultures, specifically the INS-1E and αTC-1 cell lines, which represent β and α pancreatic cells, respectively. As the pancreas is an endocrine organ, we also tested its hormone regulation capabilities. Furthermore, we assessed the secretion of MOTS-c after incubating the cells with glucose and free fatty acids. Additionally, we examined key cell culture parameters such as cell viability, proliferation, and apoptosis. The results obtained from this study show that MOTS-c has a significant impact on the physiology of pancreatic cells. Specifically, it lowers insulin secretion and expression in INS-1E cells and enhances glucagon secretion and expression in αTC-1 cells. Furthermore, MOTS-c affects cell viability and apoptosis. Interestingly, insulin and glucagon affect the MOTS-c secretion as well as glucose and free fatty acids. These experiments clearly show that MOTS-c is an important regulator of pancreatic metabolism, and there are numerous properties of MOTS-c yet to be discovered.
Impact of dietary salicylates on angiogenic factors and biochemical parameters in a rat model of preeclampsia
Background The pathophysiology of preeclampsia involves impaired cytotrophoblastic invasion, placental ischemia, inflammation, and angiogenic imbalance. Prophylactic low-dose aspirin can reduce the risk of preeclampsia and fetal growth restriction in high-risk women. This study evaluated the effect of dietary salicylates on the development of preeclampsia in rats treated with L-NAME (NG-nitro-L-arginine-methyl ester). Methodology Pregnant Sprague-Dawley rats were randomly assigned to six groups and treated with dietary salicylates at two dose levels (1 and 10 mg/kg diet) or aspirin (doses adjusted to dietary salicylates). Preeclampsia was induced by administering L-NAME in drinking water from gestational days 6–19. Results Neither dietary salicylates nor aspirin, at either dose, affected blood pressure in L-NAME-treated rats. The lower dose of dietary salicylates significantly reduced urinary albumin levels. Both interventions prevented an increase in the sFlt/PLGF ratio and mitigated histopathological placental changes in preeclamptic rats. The higher dose of aspirin reduced placental VEGFR2 protein levels. Conclusion Dietary salicylate supplementation does not provide clear preventive effects against preeclampsia.
GIP_HUMAN [22–51] Peptide Encoded by the Glucose-Dependent Insulinotropic Polypeptide (GIP) Gene Suppresses Insulin Expression and Secretion in INS-1E Cells and Rat Pancreatic Islets
GIP_HUMAN [22–51] is a recently discovered peptide that shares the same precursor molecule with glucose-dependent insulinotropic polypeptide (GIP). In vivo, chronic infusion of GIP_HUMAN [22–51] in ApoE−/− mice enhanced the development of aortic atherosclerotic lesions and upregulated inflammatory and proatherogenic proteins. In the present study, we evaluate the effects of GIP_HUMAN [22–51] on insulin mRNA expression and secretion in insulin-producing INS-1E cells and isolated rat pancreatic islets. Furthermore, we characterize the influence of GIP_HUMAN [22–51] on cell proliferation and death and on Nf-kB nuclear translocation. Rat insulin-producing INS-1E cells and pancreatic islets, isolated from male Wistar rats, were used in this study. Gene expression was evaluated using real-time PCR. Cell proliferation was studied using a BrdU incorporation assay. Cell death was quantified by evaluating histone-complexed DNA fragments. Insulin secretion was determined using an ELISA test. Nf-kB nuclear translocation was detected using immunofluorescence. GIP_HUMAN [22–51] suppressed insulin (Ins1 and Ins2) in INS-1E cells and pancreatic islets. Moreover, GIP_HUMAN [22–51] promoted the translocation of NF-κB from cytoplasm to the nucleus. In the presence of a pharmacological inhibitor of NF-κB, GIP_HUMAN [22–51] was unable to suppress Ins2 mRNA expression. Moreover, GIP_HUMAN [22–51] downregulated insulin secretion at low (2.8 mmol/L) but not high (16.7 mmol/L) glucose concentration. By contrast, GIP_HUMAN [22–51] failed to affect cell proliferation and apoptosis. We conclude that GIP_HUMAN [22–51] suppresses insulin expression and secretion in pancreatic β cells without affecting β cell proliferation or apoptosis. Notably, the effects of GIP_HUMAN [22–51] on insulin secretion are glucose-dependent.
MOTS-c modulates pancreatic islet function in rats and pigs in vitro
Abstract MOTS-c is a promising regulator of metabolism and energy homeostasis. While its effects have been studied in cell lines, our team aimed to investigate its influence on more complex structures—specifically, isolated pancreatic islets. We used two animal models: the rat, which is commonly studied, and the pig, which shares greater physiological similarities with humans. This study assessed the expression and secretion of insulin and glucagon, the expression of their receptors, cell viability, and cell death following MOTS-c treatment of the islets. Additionally, we examined how MOTS-c secretion is affected by different incubation media, such as the presence of free fatty acids, pancreatic hormones, and different glucose concentrations. The results indicate that MOTS-c impacts pancreatic islet physiology by, for example, reducing insulin and glucagon secretion and enhancing cell viability. Notably, the effects differed between the two species, which may be attributed to anatomical differences in their pancreatic islets or structural variations in rat and pig MOTS-c. These facts may lead to the conclusion that if MOTS-c may be helpful in human medicine, the pig model should be considered another valuable choice.
Effect of Paulownia Leaves Extract Levels on In Vitro Ruminal Fermentation, Microbial Population, Methane Production, and Fatty Acid Biohydrogenation
Paulownia is a fast-growing tree that produces a huge mass of leaves as waste that can be used as a feed source for ruminants. The previous study showed that phenolic compounds were the most active biological substances in Paulownia leaves, which affected the ruminal parameters and methane concentration. However, there are no scientific reports on the Paulownia leaves extract (PLE) containing phenolic compounds for their mode of action in the rumen. Phenolics constituted the main group of bioactive compounds in PLE (84.4 mg/g dry matter). PLE lowered the concentration of ammonia, modulated the VFA profile in the ruminal fluid, and decreased methane production. The PLE caused a significant reduction of in vitro dry matter degradability, reduced the number of methanogens and protozoa, and affected selected bacteria populations. PLE had a promising effect on the fatty acid profile in the ruminal fluid. Paulownia as a new dietary component or its extract as a feed additive may be used to mitigate ruminal methanogenesis, resulting in environmental protection and reducing ruminal biohydrogenation, improving milk and meat quality.
Effects of isoflavone and probiotic intake on calcium transport and bone metabolism biomarkers in female rats
AbstractCalcium is essential for maintaining bone health as it contributes to bone formation, remodeling, strength, and density. This study investigated the effect of isoflavones and probiotics on calcium transporters' gene expression, serum calcium levels, and bone metabolism biomarkers in healthy female rats. Forty‐eight female Wistar rats were classified into six groups. Bone metabolism biomarkers (pyridinoline, deoxypyridinoline, parathyroid hormone, and osteocalcin) and serum calcium levels were measured by enzyme‐linked immunosorbent assay (ELISA) and atomic absorption spectroscopy (AAS), respectively. Gene expression of calcium transporters (Trpv5 and Trpv6) was evaluated in duodenum and jejunum tissue samples using quantitative polymerase chain reaction (qPCR). Trpv5 and Trpv6, epithelial calcium channels, play a crucial role in calcium transport and homeostasis in the body. The study consisted of a1‐week adaptation period for the rats to adjust to the controlled conditions, followed by an 8‐week intervention phase. The daidzein and genistein group showed a significant increase in the gene expression of the Trpv6 transporter in the duodenum and a marked decrease in serum pyridinoline levels compared to the control group. The tempeh and soybean groups showed a significant decrease in the gene expression of the Trpv5 calcium transporter in the jejunum. However, no significant influence of the Lactobacillus acidophilus diet on calcium transport and bone metabolism biomarkers was observed in the L. acidophilus group. The correlation analysis showed a significant positive relationship between serum calcium, bone metabolism biomarkers, and calcium transporters. In conclusion, our study demonstrates that the daidzein and genistein diet improves calcium transport in the duodenum and reduces pyridinoline serum concentrations, while tempeh and soybean diets reduce calcium transport in the jejunum. However, the combination of daidzein, genistein, and L. acidophilus did not demonstrate a synergistic effect on calcium transport and bone metabolism, suggesting that further investigations are needed to elucidate their potential interactions.
Rice Bran in Old Horses Nutrition and its Influence on Condition, Blood Biochemical Parameters, Total Feces Bacteria and Methanogen Population
Abstract This study aimed to verify whether the inclusion of 0.5 kg full-fat rice bran per day in the diet of geriatric horses will improve their condition, increase the population of methanogens in the cecum, and thus affect the biochemical blood parameters. The experiment included 2 research periods: 6 healthy, non-working horses over 20 years of age (480 ± 20 kg of body weight) fed only hay (±8.86 kg/day/head) in the first period and hay (±8.00 kg/day/head) and rice bran (0.5 kg/day/head) in the second one. Each of these periods lasted 4 months. The Body Condition Scoring (BCS) assessment was performed at the beginning and end of the experiment. Blood and feces samples were collected on the first and last day of each period. After feeding with the addition of rice bran, BCS increased by 1.17 units on a 9-point scale. The experiment showed an increase in the total number of bacteria and methanogens inhabiting the cecum of horses. This can lead to better digestion of carbohydrates, absorption of nutrients, and, consequently, increased body weight. No differences occurred in the hematology and serum biochemistry indices of horses fed a diet including rice bran, except for the amount of serum globulin and the albumin to globulin ratio. Rice bran affected essential serum fatty acid profile (increased PUFA and decreased MUFA), which confirmed the possibility to use diet as a serum fatty acids profile modulator.
Effect of clinically uneventful carotid endarterectomy on pentraxin 3 concentration: relationships between cognitive functions, local inflammation, and duration of brain ischemia
IntroductionThe pentraxins are a superfamily of proteins characterized by the presence of a pentraxin domain at the C-terminal. One of them is pentraxin 3 (PTX3), which is of great interest to scientists, primarily due to its function as an endogenous modulator of the inflammatory response. Local inflammation is a common phenomenon during various types of surgery and procedures, and its causes, among others, are local ischemia or tissue irritation. One such procedure is carotid plaque removal by carotid endarterectomy (CEA), which is performed to reduce the risk of stroke.Material and methodsThe purpose of this study was to investigate the effect of CEA on PTX3 concentration and the relationship between cognitive functions and the duration of brain ischemia in patients. We evaluated the relationships using blood collected from patients undergoing CEA, the Mini-Mental State Examination (MMSE) questionnaire, and commercially available ELISA tests.ResultsResults: We found that the concentration of PTX3 increased after surgery (from 0.59 ± 0.11 to 1.29 ± 0.29 ng mL<sup>–1</sup>; <i>P</i> < 0.01). Also, a decrease in the MMSE score was observed 24 hours after the procedure (<i>P</i> < 0.01). However, just one month after the procedure, it significantly increased compared to the day of admission to the hospital (<i>P</i> < 0.01). Our results did not demonstrate any interactions between the tested parameters. It can be concluded that neither the changes in PTX3 concentration following the procedure nor the timing of these changes directly contribute to the temporary deterioration of cognitive functions observed after CEA.ConclusionsPTX3 increases independently of the duration of cerebral hypoxemia/ hypercapnia after clamping the carotid artery. This rise is an inflammatory marker that occurs independently within the central nervous system.
Structural and biomechanical analysis of femurs from mice treated with diclofenac, miR-15b and miR-365
PurposeNonsteroidal anti-inflammatory drugs (NSAIDs) are used to treat pain, but they have side effects, including the inhibition of bone healing. Diclofenac (DF), a member of the NSAID group, affects bone health adversely. One potential approach to protect bones from the effects of NSAIDs involves the administration of short nucleic acids, such as microRNAs (miRNAs). This study aimed to determine whether two specific miRNAs, miR-15b and miR-365, could mitigate the effects of DF on bone.MethodsWe used the C57BL/6J mouse strain and the MC3T3-E1 pre-osteoblast cell line derived from this mouse strain. Female C57BL/6J mice were treated with DF and miR-15b or miR-365 mimics. After euthanising the mice, we analysed their femurs using micro-computed tomography (μCT) and dynamic mechanical analysis (DMA). In addition, we performed experiments in cultured MC3T3-E1 cells, which were transfected with either miR-15b or miR-365. We assessed the relative mRNA levels of osteoblast differentiation markers using real-time PCR.ResultsOur findings indicated that miR-15b and miR-365 were effective in reversing the detrimental effects of DF on bone mineral density. DF decreased the bone’s storage modulus (E’), while miR-15b and miR-365 ameliorated this effect. In the preosteoblast MC3T3-E1 cells, DF did not significantly regulate marker genes; however, the administration of miR-15b and miR-365 reduced the gene expression of Runx2, Alp, and Satb2.ConclusionsIn summary, the impact of DF on the structural and mechanical properties of bone was not mediated by gene regulation in osteoblasts. However, osteoblasts were responsive to the administration of miR-15b and miR-365.
Implementation of a metabolizable energy regression model for black soldier fly larvae fat in broiler chicken diets: effect on growth performance, nutrient digestibility, and selected physiological indices
LEAP2 in Physiology—A Narrative Review
Liver Enriched Antimicrobial Peptide 2 (LEAP2) is a fascinating peptide that has gained significant attention since its discovery in 2003. Initially identified as an antimicrobial peptide, LEAP2 has more recently been found to play a key role in the regulation of energy metabolism. One of the most notable functions of LEAP2 is its interaction with the ghrelin hormone, which is known for stimulating hunger. LEAP2 acts as an inhibitor of ghrelin, thereby reducing food intake and influencing energy balance. The physiological roles of LEAP2 extend beyond appetite suppression. Studies have shown that LEAP2 has an impact on insulin secretion, suggesting its potential involvement in glucose metabolism and possibly insulin sensitivity, which is crucial in managing conditions like type 2 diabetes. Moreover, LEAP2 levels appear to fluctuate based on factors such as gender, developmental stage, and even interventions like bariatric surgery, which is known for its role in managing obesity and diabetes. Given these findings, LEAP2 shows potential as a therapeutic target, particularly for addressing obesity and metabolic diseases such as type 2 diabetes. Its ability to influence food intake and energy balance makes it a promising candidate for further research into therapies aimed at weight regulation and glycemic control. In the future, LEAP2 could become an important agent in the development of treatments aimed at curbing obesity and its associated metabolic disorders.
Cord Blood Spexin Level in Mothers with Obesity—Forecast of Future Obesity?
Spexin (SPX) is a peptide that plays an important role in the regulation of food intake and body weight (BW) by the effect on carbohydrate-lipid metabolism. However, the role of SPX in fetal life, in children, and in adolescent metabolism is limited. Therefore, we decided to check whether obesity affects the concentration of SPX in the mother’s peripheral blood (MB) and umbilical cord blood (UCB). Using MB and UCB sera on the day of delivery obtained from 48 women (24 non-obese and 24 obese) and commercially available Elisa kits and colorimetric assays, we determined changes in SPX and the relationship between SPX concentration and other metabolic and anthropometric markers (body weight and BMI) on the day of delivery and in children at the age of 36 months. We found lower concentrations of SPX in MB (p < 0.05) and UCB (p < 0.01) derived from obese women (BMI > 30) and a moderate linear correlation (r = 0.4429; p < 0.01) between SPX concentrations in MB and UCB. We also noted that the concentration of SPX is not correlated with the child’s body weight on the day of birth (r = −0.0128). However, there is a relationship between SPX at birth and body weight at 3 years of age (r = −0.3219; p < 0.05). Based on the obtained results, it can be assumed that spexin is one of the factors modulating the child’s metabolism already in the fetal period and can be considered a potential marker of future predisposition to obesity. However, confirmation of this thesis requires additional research.
