Search Results (332)

Background: In the bovine mammary gland, de novo fatty acid synthesis is a critical process for milk fat production, in which acetyl-CoA synthetase 2 (ACSS2) serves as a key enzyme by converting acetate into acetyl-CoA. This metabolic pathway is intricately regulated by non-coding RNAs, particularly through the competitive endogenous RNA (ceRNA) mechanism.Purpose: To elucidate the regulatory role and molecular mechanism of the circRNA-02213/miR-328/ACSS2 axis in the lipid metabolism of bovine mammary epithelial cells (BMECs). Methods: Bioinformatic prediction and dual-luciferase reporter assays were employed to verify the targeting interactions among circRNA-02213, miR-328, and ACSS2. In BMECs, qRT-PCR, Western blot, triglyceride/cholesterol quantification, Oil Red O staining, and cell proliferation assays were used to evaluate the effects of this axis on key lipid-metabolic indices and cellular phenotypes. Results: circRNA-02213 functioned as a molecular “sponge” that sequestered miR-328, thereby upregulating ACSS2 expression. Functionally, circRNA-02213 overexpression markedly promoted triglyceride and cholesterol synthesis, lipid droplet accumulation, and BMEC proliferation; whereas miR-328 exerted significant inhibitory effects on these lipid-metabolic processes and cell proliferation. Conclusions: This study demonstrates that circRNA-02213 acts as a ceRNA to relieve miR-328-mediated repression of ACSS2, constituting a critical network that regulates milk fat synthesis and metabolism. The circRNA-02213/miR-328/ACSS2 axis represents a potential molecular target for improving milk lipid quality in ruminants. Full article

Background: Degenerin proteins, such as Acid-Sensing Ion Channel 2 (ASIC2) and β Epithelial Na⁺ Channel (βENaC), have been implicated in cardiovascular function. We previously demonstrated that mice lacking normal levels of βENaC and ASIC2 are protected from diet-induced obesity, metabolic disruption, and hepatic steatosis. Methods: To investigate the specific role of βENaC proteins in the progression of metabolic disease, we examined the impact of a high-fat diet (HFD) in the βENaC hypomorph mouse model (βMUT). Body composition and metabolic and behavioral phenotypes were examined in male and female and βMUT and WT mice (n = 6–14/group) fed a normal chow diet (NFD) from weaning until 16 weeks of age, then a 60% kcal-fat diet for 5 weeks. Results: Compared to WT mice, βMUT male mice have reduced lean and total body mass. No remarkable differences in energy expenditure, motor activity, or food consumption patterns were detected. HFD-fed male βMUT mice exhibited reduced liver fat content (mass and Oil Red O staining) yet increased abdominal fat depots. HFD-fed female βMUT mice exhibited lower heart mass. Conclusions: These novel findings suggest a role for βENaC in the maintenance of metabolic homeostasis and adipose tissue distribution. Full article

Kratom (Mitragyna speciosa (Korth.) Havil.) is a medicinal plant containing bioactive alkaloids, notably mitragynine and 7-hydroxymitragynine, which are psychoactive compounds with analgesic and stimulant properties. Due to safety concerns, the use of Kratom leaves and mitragynine in oral pharmaceutical products is restricted. Therefore, their potential as topical cosmeceutical agents merits further exploration. This study aimed to investigate the antioxidant and anti-adipogenic activities of Kratom ethanolic (Et-MS) and alkaloid-rich (Alk-MS) extracts, as well as purified mitragynine, to determine whether mitragynine is the major bioactive compound responsible for lipid reduction in 3T3-L1 adipocytes. The antioxidant properties were assessed using DPPH, ABTS, and FRAP assays, yielding EC₅₀ values of 0.06 mg/mL, 0.29 mg/mL, and 55 g Fe²⁺/100 g for Et-MS, respectively. In comparison, ascorbic acid (positive control) showed a DPPH EC₅₀ value of 0.002 mg/mL. Both Alk-MS and mitragynine significantly inhibited lipid accumulation in 3T3-L1 adipocytes by up to 50–70% at non-cytotoxic concentrations (≤25 µg/mL), as determined by Oil Red O staining. These findings provide preliminary in vitro evidence that phenolic constituents contribute to antioxidant capacity, while mitragynine is the principal anti-adipogenic constituent in Kratom extracts. Collectively, the results support the potential for further development of Kratom-derived extracts and mitragynine as plant-based candidates for topical or cosmeceutical applications targeting subcutaneous fat and oxidative skin damage. Full article

Background: Exercise interacts closely with the circadian system; however, whether long-term training elicits time-of-day-dependent metabolic adaptations in the context of obesity remains unclear. Methods: Male C57BL/6 mice were fed a high-fat diet and trained on a treadmill for 8 weeks during either the early rest phase (ZT3, Zeitgeber time) or the early active phase (ZT15). Sedentary mice served as controls. After the last session, animals were fasted for 4 h and sampled 48–49 h later. Plasma triglycerides (TGs) and glucose, as well as liver and epididymal white adipose tissue (EPI), were analyzed. Results: Plasma TGs showed a significant phase × exercise interaction (F(1, 25) = 5.25, p = 0.0307), with the lowest levels in ZT15-exe (27.22 mg/dL) compared with ZT15-sed (39.47 mg/dL, p < 0.01) and ZT3-exe (41.80 mg/dL, p < 0.01). Hepatic TG content was markedly lower in ZT3 than in ZT15 mice (F(1, 25) = 15.49, p < 0.001), and Oil Red O staining was associated with reduced lipid accumulation in exercised groups (p < 0.05). In EPI, Fasn expression was robustly decreased by exercise (F(1, 25) = 16.43, p = 0.0004, q = 0.0059), indicating long-term suppression of lipogenesis. In the liver, Cpt1a showed significant main effects of both phase (F(1, 25) = 10.11, p = 0.0039, q = 0.0158) and exercise (F(1, 25) = 13.42, p = 0.0012, q = 0.0353), being higher in ZT3 and under sedentary conditions, suggesting a circadian-dominant oxidative advantage in hepatic metabolism. Conclusions: Long-term exercise induced phase-dependent adaptations in lipid metabolism. Active-phase exercise promoted adipose lipid mobilization and lowered plasma TGs, while rest-phase training enhanced hepatic oxidative capacity. These results suggest a “tissue × time” framework of circadian-specific exercise responses, providing hypothesis-generating evidence for optimizing exercise timing in metabolic disorders. Full article

Background: As a key contributor to metabolic disorders, obesity is recognized as a critical global health challenge. Adipocyte differentiation depends on the mitotic clonal expansion (MCE) phase, which is controlled by oxidative balance and transcription factors like C/EBPβ. Sesaminol, a lignan derived from Sesamum indicum, has potent antioxidant properties. This study aimed to investigate whether sesaminol suppresses adipogenesis by modulating ROS signaling, MCE, and the Nrf2-ARE pathway. Methods: In the early period of adipogenic induction, 3T3-L1 preadipocytes received treatment with sesaminol. Adipogenic development was evaluated through Oil Red O staining together with the assay of GPDH activity. Assays of cell proliferation and expression of cell cycle-related proteins, along with ROS measurement, qRT-PCR, Western blotting, and immunofluorescence, were performed to evaluate the effects on oxidative stress, transcriptional regulation, and AMPK-Nrf2 signaling. Results: Sesaminol significantly inhibited lipid accumulation and GPDH activity without cytotoxicity. It suppressed MCE by inhibiting DNA synthesis and reducing the expression of cyclin E1/E2 and CDK2. Sesaminol decreased C/EBPβ expression and its nuclear localization, resulting in lower levels of C/EBPα and PPARγ. It also reduced intracellular ROS, promoted nuclear translocation of Nrf2, and upregulated antioxidant genes HO-1 and GCLC. AMPK phosphorylation was concurrently enhanced. Conclusions: Sesaminol inhibits early adipogenesis by suppressing ROS-mediated MCE and activating the AMPK-Nrf2-ARE signaling pathway, leading to downregulation of key adipogenic transcription factors. The present study supports the potential of sesaminol as an effective strategy for obesity prevention. Full article

Background: Brown adipose tissue (BAT) is indispensable for producing heat and contributes critically to the survival of neonatal mammals. MicroRNAs (miRNAs) are small noncoding RNAs that serve as key post-transcriptional regulators, playing a crucial role in regulating BAT development and thermogenesis. However, the role of miR-326-3p in goat brown adipocytes remains largely unclear. Methods: Primary brown adipocytes were isolated from goat perirenal adipose tissue and subjected to gain and loss-of-function assays using miR-326-3p mimics and inhibitors. Lipid accumulation, thermogenic-related genes, and mitochondrial gene expression were quantified by Oil Red O staining and qRT-PCR. Target prediction and dual-luciferase reporter assays were performed to validate direct interaction between miR-326-3p and FGF11. Results: Expression profiling demonstrated that miR-326-3p is more enriched in brown adipose tissue (BAT) than in white adipose tissue (WAT), and the expression level gradually decreases with adipocyte differentiation. miR-326-3p overexpression significantly inhibited lipid droplet accumulation and the expression of genes associated with differentiation, thermogenesis, and mitochondria, including PPARγ, FABP4, UCP1, and PGC1α, whereas inhibition produced the opposite effect. Bioinformatic prediction and dual-luciferase reporter assays further identified fibroblast growth factor 11 (FGF11) as a direct target of miR-326-3p. Conclusions: These findings reveal that miR-326-3p negatively regulates the differentiation and expression of thermogenic-related genes of goat brown adipocytes, uncovering a novel miR-326-3p-FGF11 regulatory axis. Full article

Background/Objectives: Obesity, defined by the excessive accumulation of adipose tissue, is associated with an increased risk of type 2 diabetes and metabolic dysfunction-associated steatotic liver disease (MASLD). Obesity treatments based on natural products are receiving increasing attention as viable alternatives to conventional treatments. Methods: To investigate the anti-obesity effects of Paliurus ramosissimus leaf extract (PRLE) in vitro and in vivo, we conducted studies using 3T3-L1 pre-adipocytes. The in vivo studies used high-fat diet (HFD)-fed C57BL/6 mice. PRLE effects were assessed through Oil Red O staining, RT-qPCR, Western blot, and morphological analysis of adipose tissue. Results: PRLE significantly reduced lipid accumulation in 3T3-L1 cells without cytotoxicity. PRLE treatment decreased mRNA expression of adipogenic genes (PPARγ, C/EBPα, FABP4, and leptin) and protein levels of adipogenesis-related markers. In HFD-fed mice, PRLE administration significantly reduced body weight gain (p < 0.001), decreased adipose tissue mass, and diminished the weight and size of white adipose tissue. Conclusions: PRLE exhibits anti-obesity effects both in vitro and in vivo, suggesting its potential as a therapeutic agent for obesity prevention. Full article

Objectives: This study focuses on the regulatory mechanism of Schisandrin B (Sch B) on the lipid metabolism and apoptosis of AML-12 liver cells, with a particular emphasis on its potential therapeutic effect and mechanism of action in preventing and treating metabolic-associated fatty liver disease (MAFLD) by activating the PPARγ signaling pathway. Methods: An MAFLD cell model was established by inducing AML-12 cells with a mixture of oleic acid (OA) and palmitic acid (PA) (2:1). AML-12 cells were divided into a control group, a model group, and 20 μM and 40 μM Sch B groups. The cells were lysed and prepared into the cell suspension, then the cell suspension was centrifuged to obtain its supernatant, and the levels of total cholesterol (TC), triglycerides (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in the supernatant were detected according to the instructions of the kits. Effects of Sch B on the pathological changes of AML-12 cells were observed by Oil Red O staining. The key targets were screened through network pharmacology, and relevant targets were verified through molecular docking simulation. The activity of PPARγ was detected using a dual luciferase reporter plasmid, and the level of cell apoptosis was detected using the Annexin V-FITC/PI double staining method. The Western blot method was used to analyze the expression of genes related to lipid metabolism and apoptosis pathways. Results: Sch B could regulate lipid metabolism disorders in OA+PA-induced MAFLD cell model. The activation of PPARγ-PCK1/Aspase is a key step in the action of Sch B, which can effectively block fatty acid synthesis, improve fatty acid oxidation, and reduce lipid droplet aggregation in liver cells, thereby alleviating lipid metabolism abnormalities in the MAFLD cell model and inhibiting cell apoptosis. Conclusions: This finding may lay an important theoretical foundation and open a new research direction for the deep development and application of Schisandra chinensis. Full article

Background: Hexavalent chromium (Cr(VI)) can migrate into soil and water, posing risks to animal health. However, it remains unclear whether Cr(VI) perturbs essential trace elements and antioxidant gene expression, triggers apoptosis, or disrupts hepatic lipid metabolism in New Zealand rabbits. Methods: To address this knowledge gap, twenty-four 30-day-old New Zealand rabbits were randomly allocated to one control and three Cr(VI)-treated groups (differing in Cr(VI) concentration) and maintained for 28 days. Livers were then harvested for analysis. Total Cr and essential trace elements were quantified by ICP-OES. Hematoxylin–eosin staining and transmission electron microscopy were employed to assess histopathological and ultrastructural alterations, respectively. Hepatic lipid accumulation was visualized with Oil Red O staining. QRT-PCR was used to determine the expression of antioxidant and lipid-metabolism-related genes. Results: Cr(VI) was detectable in liver tissue at all exposure levels and was accompanied by significant decreases in four essential trace elements (Fe, Mn, Zn, and Se); Cu displayed a biphasic response, rising at lower Cr(VI) doses before declining at higher doses. Histopathological and ultrastructural analyses revealed overt hepatic injury. Notably, all Cr(VI) treatments elevated antioxidant gene expression, indicating activation of hepatic defense pathways. Lipid metabolism was also disrupted, evidenced by increased lipid deposition and up-regulation of genes governing hepatic fat metabolism. Conclusions: Collectively, these findings demonstrate that Cr(VI) elicits dose-dependent activation of hepatic antioxidant defenses, promotes apoptosis, and induces lipid-metabolic disorders in New Zealand rabbit hepatocytes. This study provides novel mechanistic insights into Cr(VI)-induced hepatotoxicity and offers a valuable reference for evaluating the hepatic risks of environmental Cr(VI) exposure in this species. Full article

Background: Primary hepatocytes are excellent models for studying liver functions and liver diseases. However, obtaining high yields of viable hepatocytes remains technically challenging, limiting their broader applications. Most conventional methods rely on a two-step collagenase perfusion technique. Despite its widespread use, this approach has several limitations that reduce the success rate of hepatocyte isolation and culture. The procedure involves multiple parameters that are continually being optimized in order to obtain hepatocytes in high yield and quality that can be used to provide insights into their physiology and pathophysiology. Aim: We aimed to enhance the success rate and reproducibility of hepatocyte isolation with high yield, enabling analysis of diverse physiological and pathophysiological aspects of lipid metabolism. It also establishes an in vitro steatosis model for evaluating therapeutic drugs and molecular interventions. Methods: Rat liver was perfused in situ with EDTA buffer followed by collagenase IV. Liver was then isolated, and hepatocytes were mechanically liberated, filtered, and purified through density-gradient centrifugation. Viable cells were cultured at 700,000 or 1 million cells/well for 24 h. The monolayer was incubated in lipogenic media for an additional 24 or 48 h. Hepatocytes were fixed, neutral lipids were stained using Oil Red O, and the stained area was quantified using Image J software version 1.54. Results: Yield of hepatocytes was ~75–90 million cells/liver, with viability of 86–93%. Cells seeded at 700,000 and 1 million cells/well reached confluences of 60% and 80%, respectively, after 24 h. Steatosis was then induced with lipid accumulation reaching 21% of image area after 24 h and 25% after 48 h. Conclusions: The current protocol presents an efficient and highly reproducible method for isolating primary rat hepatocytes in high yield with high viability. Additionally, the protocol provides a foundation for studying the pathophysiology of fatty liver disease. Full article

Background: Fat embolism frequently occurs as a result of trauma, such as long bone fractures and orthopedic surgeries, as well as in certain non-traumatic conditions. The formation can be attributed to mechanical or biochemical processes. According to Hullman’s biochemical hypothesis, elevated C-reactive protein levels facilitate the precipitation of very-low-density lipoproteins and chylomicrons, forming fat globules that may result in fat embolism. Based on the abovementioned hypothesis, this study aims to detect fat embolism in autopsy patients (postmortem) suffering from bronchopneumonia and determine its possible role as a cause of death. Methods: A group of autopsies of deceased individuals with bacterial purulent bronchopneumonia with confirmed or presumed elevated C-reactive protein levels was rigorously selected, excluding those with other potential causes of fat embolism such as cardiopulmonary resuscitation, hypothermia, and diabetes mellitus. Multiple organs were sampled for frozen section analysis using Oil Red O fat staining and assessed for the presence and extent of fat embolism. The Falzi score, as modified by Janssen, was employed for the lung tissue. Results: In 73% of the cases, predominantly sporadic, Grade 0 or Grade I fat embolism was observed; however, in none of the cases was fat embolism identified as the cause of death or as a significant contributing factor. Furthermore, neither fat embolism syndrome nor multiorgan fat embolism were detected. Conclusions: Although an elevated C-reactive protein level facilitates the formation of fat globules and fat embolism, its role as a direct cause of mortality remains uncertain. It may predispose individuals to such conditions and potentially interact with other factors, such as minor soft tissue trauma, to exacerbate the severity of fat embolism or its clinical manifestations. These findings underscore the necessity for further comprehensive investigations within the contexts of infection/inflammation, fat embolism, and dyslipidemia. Full article

Lipofilling has far more applications than cosmetic surgery alone. Due to its high content of stromal vascular fraction (SVF) cells, lipoaspirate can also be used to treat wounds, as its cellular components may accelerate wound healing. Using our CELT^PLUS protocol, we can increase the number of SVF cells per volume. Unfortunately, some patients require more than one treatment to achieve an optimal outcome, but would unnecessarily suffer from repeated liposuction. Therefore, our objective was to test whether cryopreserving CELT^PLUS fat could offer a solution, potentially avoiding the need for repeated liposuction procedures. DMSO was used as a cryoprotective agent for proof-of-principle testing, although other non-toxic cryoprotective agents should be considered in the future. The rest of our freezing protocol is a clinically friendly attempt to facilitate the translation into clinical practice. We tested the cryopreserved tissue using histological evaluation, metabolism measurement, SVF cell yield estimation, PCRs from both whole tissue and from cultured SVF cells, and Oil Red “O” staining. We found that freezing CELT^PLUS fat with DMSO yields better results than without cryoprotection in all evaluated methods. Until non-toxic cryoprotective agents are tested on CELT^PLUS fat, we do not recommend initiating animal or human testing. Full article

Sebum secreted by sebaceous glands mixes with sweat to form a protective film that aids in maintaining skin health. Reduced sebum production compromises such barrier functions, potentially leading to severe itchiness and inflammation. Therefore, incorporating moisturizers with ingredients promoting sebum secretion is desirable. Wild watermelon possesses moisturizing and antioxidant properties, and its extracts are utilized in skin cosmetics and supplements. This study investigates whether seed watermelon (Citrullus mucosospermus (Fursa))—a species closely related to wild watermelon—influences sebum synthesis and can serve as a skin cosmetic raw ingredient. Several bioactive compounds—including coniferyl alcohol, coniferin, and p-coumaryl alcohol—were identified in the active third fraction of the fruit extract. Subsequently, SZ95 sebocytes stimulated with linoleic acid were stained using Oil Red O to detect lipogenesis facilitated by the identified bioactive compounds. Coniferyl alcohol promoted linoleic acid-stimulated lipogenesis by approximately 2.2-fold at a concentration of 300 µM. Lipidomic analyses confirmed an increase in total lipid content following coniferyl alcohol treatment, with notable increases in cholesterol ester, cardiolipin, and simple lipid content. Overall, these findings suggest that seed watermelon contains compounds that do influence sebum synthesis. Consequently, skin cosmetics containing seed watermelon fruit extracts with linoleic acid may benefit individuals with dry skin. Full article

Background: Cucurbitacin E (CuE), a natural tetracyclic triterpenoid compound extracted from the melon stems of Cucurbitaceae plants, has been reported to exhibit anti-inflammatory and anti-cancer properties, along with the ability to enhance cellular immunity. However, its role and molecular mechanism in regulating lipid metabolism and adipogenesis remain unclear. This study aims to investigate the potential anti-adipogenic and anti-obesity effects of CuE in 3T3-L1 adipocytes. Materials and Methods: 3T3-L1 preadipocytes were cultured and induced to differentiate using a standard adipogenic cocktail containing dexamethasone, 3-isobutyl-1-methylxanthine (IBMX), and insulin (DMI). CuE was administered during the differentiation process at various concentrations. Lipid accumulation was assessed using Oil Red O staining, and cell viability was evaluated via the MTT assay. To determine whether CuE induced apoptosis or necrosis, flow cytometry was performed using annexin V/PI staining. Additional molecular analyses, such as Western blotting and RT-PCR, were used to examine the expression of key adipogenic markers. Results: Treatment with CuE significantly reduced lipid droplet formation in DMI-induced 3T3-L1 adipocytes in a dose-dependent manner, as shown by decreased Oil Red O staining. Importantly, CuE did not induce apoptosis or necrosis in 3T3-L1 cells at effective concentrations, indicating its safety toward normal adipocytes. Moreover, CuE treatment downregulated the expression of adipogenic markers such as PPARγ and C/EBPα at both mRNA and protein levels. Discussion: Our findings suggest that CuE exerts a non-cytotoxic inhibitory effect on adipocyte differentiation and lipid accumulation. This anti-adipogenic effect is likely mediated through the suppression of key transcription factors involved in adipogenesis. The absence of cytotoxicity supports the potential application of CuE as a safe bioactive compound for obesity management. Further investigation is warranted to elucidate the upstream signaling pathways and in vivo efficacy of CuE. Conclusions: Cucurbitacin E effectively inhibits adipogenesis in 3T3-L1 adipocytes without inducing cytotoxic effects, making it a promising candidate for the development of functional foods or therapeutic agents aimed at preventing or treating obesity. This study provides new insights into the molecular basis of CuE’s anti-obesity action and highlights its potential as a natural lipogenesis inhibitor. Full article

Background: Alcohol-associated liver disease (ALD) is a primary global health concern, exacerbated by oxidative stress, inflammation, and gut barrier dysfunction. Conventional phytocompounds exhibit hepatoprotective potential but are hindered by low bioavailability. This study aimed to evaluate the hepatoprotective and gut-barrier-restorative effects of green-synthesized silver nanoparticles (AgNPs) derived from Enicostemma littorale, a medicinal plant known for its antioxidant and anti-inflammatory properties. Methods: AgNPs were synthesized using aqueous leaf extract of E. littorale and characterized using UV-Vis, XRD, FTIR, DLS, and SEM. HepG2 (liver) and Caco-2 (colon) cells were exposed to 0.2 M ethanol, AgNPs (1–100 µg/mL), or both, to simulate ethanol-induced toxicity. A range of in vitro assays was performed to assess cell viability, oxidative stress (H₂DCFDA), nuclear and morphological integrity (DAPI and AO/EtBr staining), lipid accumulation (Oil Red O), and gene expression of pro- and anti-inflammatory, antioxidant, and tight-junction markers using RT-qPCR. Results: Ethanol exposure significantly increased ROS, lipid accumulation, and the expression of inflammatory genes, while decreasing antioxidant enzymes and tight-junction proteins. Green AgNPs at lower concentrations (1 and 10 µg/mL) restored cell viability, reduced ROS levels, preserved nuclear morphology, and downregulated CYP2E1 and SREBP expression. Notably, AgNPs improved the expression of Nrf2, HO-1, ZO-1, and IL-10, and reduced TNF-α and IL-6 expression in both cell lines, indicating protective effects on both liver and intestinal cells. Conclusions: Green-synthesized AgNPs from E. littorale exhibit potent hepatoprotective and gut-barrier-restoring effects through antioxidant, anti-inflammatory, and antilipidemic mechanisms. These findings support the therapeutic potential of plant-based nanoparticles in mitigating ethanol-induced gut–liver axis dysfunction. Full article