In Vitro Assessment of Anti-Adipogenic and Anti-Inflammatory Properties of Black Cumin (Nigella sativa L.) Seeds Extract on 3T3-L1 Adipocytes and Raw264.7 Macrophages.

Background and Objectives: This study evaluated the in vitro anti-adipogenic and anti-inflammatory properties of black cumin (Nigella sativa L.) seed extract (BCS extract) as a potential candidate for developing herbal formulations targeting metabolic disorders. Materials and Methods: We evaluated the BCS extract by assessing its 2,2-diphenyl-1-picrohydrazyl (DPPH) radical scavenging activity, levels of prostaglandin E2 (PGE2) and nitric oxide (NO), and mRNA expression levels of key pro-inflammatory mediators. We also quantified the phosphorylation of nuclear factor kappa light chain enhancer of activated B cells (NF-κB) and mitogen-activated protein kinases (MAPK) signaling molecules. To assess anti-adipogenic effects, we used differentiated 3T3-L1 cells and BCS extract in doses from 10 to 100 µg/mL. We also determined mRNA levels of key adipogenic genes, including peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/BEPα), adipocyte protein 2 (aP2), lipoprotein lipase (LPL), fatty acid synthase (FAS), and sterol-regulated element-binding protein 1c (SREBP-1c) using real-time quantitative polymerase chain reaction (qPCR). Results: This study showed a concentration-dependent DPPH radical scavenging activity and no toxicity at concentrations up to 30 µg/mL in Raw264.7 cells. BCS extract showed an IC50 of 328.77 ± 20.52 µg/mL. Notably, pre-treatment with BCS extract (30 µg/mL) significantly enhanced cell viability in lipopolysaccharide (LPS)-treated Raw264.7 cells. BCS extract treatment effectively inhibited LPS-induced production of PGE2 and NO, as well as the expression of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS), interleukin (IL)-1ß and IL-6, possibly by limiting the phosphorylation of p38, p65, inhibitory κBα (I-κBα), and c-Jun N-terminal kinase (JNK). It also significantly attenuated lipid accumulation and key adipogenic genes in 3T3-L1 cells. Conclusions: This study highlights the in vitro anti-adipogenic and anti-inflammatory potential of BCS extract, underscoring its potential as a promising candidate for managing metabolic disorders.

Anti-Osteoarthritic Effects of Antarctic Krill Oil in Primary Chondrocytes and a Surgical Rat Model of Knee Osteoarthritis.

Osteoarthritis (OA) is characterized by progressive cartilage destruction and synovitis; however, there are no approved disease-modifying OA drugs. Krill oil (KO) has been reported to possess anti-inflammatory properties and alleviate joint pain in knee OA, indicating its potential to target the inflammatory mechanism of OA. Therefore, the anti-OA effects of KO were investigated in primary chondrocytes and a surgical rat model of knee OA. The oral administration of KO at 200 and 100 mg/kg for 8 weeks improved joint swelling and mobility in the animal model and led to increased bone mineral density and compressive strength in the cartilage. The oral KO doses upregulated chondrogenic genes (type 2 collagen, aggrecan, and Sox9), with inhibition of inflammation markers (5-lipoxygenase and prostaglandin E2) and extracellular matrix (ECM)-degrading enzymes (MMP-2 and MMP-9) in the cartilage and synovium. Consistently, KO treatments increased the viability of chondrocytes exposed to interleukin 1α, accompanied by the upregulation of the chondrogenic genes and the inhibition of the ECM-degrading enzymes. Furthermore, KO demonstrated inhibitory effects on lipopolysaccharide-induced chondrocyte inflammation. Histopathological and immunohistochemical analyses revealed that KO improved joint destruction and synovial inflammation, probably due to the anti-inflammatory, anti-apoptotic, and chondrogenic effects. These findings suggest the therapeutic potential of KO for knee OA.

Krill oil inhibited adipogenic differentiation by inducing the nuclear Nrf2 expression and the AMPK activity.

The current study investigated the antiadipogenic mechanism of krill oil from the 3T3-L1 adipocytes. The krill oil adhered to the criteria as a food standard by showing 50.8% of the total phospholipid, 5.27% myristic acid, and 1.63% linoleic acid. The lipid accumulation that was measured in the 3T3-L1 cells using oil red O staining was reduced up to 54% by the krill oil. The krill oil treatment reduced the adipogenic transcription factors by downregulating the sterol regulatory element binding protein 1 (SREBP1) and acetyl-CoA carboxylase (ACC), phospho-ACC, and AMP-activated protein kinase (AMPK) phosphorylation. The current study confirmed that the krill oil inhibited adipogenesis by downregulating SREBP1 and ACC via the upregulation of the AMPK and nuclear factors E2-related factor 2 (Nrf2) signaling pathway in the 3T3-L1 adipocytes. These findings suggest that krill oil is a good source of phospholipid and phosphatidylcholine, which could be a potential natural antiobesity ingredient by inhibiting adipogenesis.

Krill Oil's Protective Benefits against Ultraviolet B-Induced Skin Photoaging in Hairless Mice and In Vitro Experiments.

Krill oil (KO) shows promise as a natural marine-derived ingredient for improving skin health. This study investigated its antioxidant, anti-inflammatory, anti-wrinkle, and moisturizing effects on skin cells and UVB-induced skin photoaging in hairless mice. In vitro assays on HDF, HaCaT, and B16/F10 cells, as well as in vivo experiments on 60 hairless mice were conducted. A cell viability assay, diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity test, elastase inhibition assay, procollagen content test, MMP-1 inhibition test, and hyaluronan production assay were used to experiment on in vitro cell models. Mice received oral KO administration (100, 200, or 400 mg/kg) once a day for 15 weeks and UVB radiation three times a week. L-Ascorbic acid (L-AA) was orally administered at 100 mg/kg once daily for 15 weeks, starting from the initial ultraviolet B (UVB) exposures. L-AA administration followed each UVB session (0.18 J/cm2) after one hour. In vitro, KO significantly countered UVB-induced oxidative stress, reduced wrinkles, and prevented skin water loss by enhancing collagen and hyaluronic synthesis. In vivo, all KO dosages showed dose-dependent inhibition of oxidative stress-induced inflammatory photoaging-related skin changes. Skin mRNA expressions for hyaluronan synthesis and collagen synthesis genes also increased dose-dependently after KO treatment. Histopathological analysis confirmed that krill oil (KO) ameliorated the damage caused by UVB-irradiated skin tissues. The results imply that KO could potentially act as a positive measure in diminishing UVB-triggered skin photoaging and address various skin issues like wrinkles and moisturization when taken as a dietary supplement.

Efficacy Confirmation Test of Black Cumin (Nigella sativa L.) Seeds Extract Using a High-Fat Diet Mouse Model.

To deal with the adverse effects associated with the use of currently available treatments for metabolic disorders, such as type 2 diabetes, there is a need to find an alternative drug compound. In the present study, we investigated the therapeutic potential of black cumin (Nigella sativa L.) seeds extract (BCS extract) for type 2 diabetes using a 45% Kcal-fed obese mouse model. The BCS extract at different doses (400-100 mg/kg) showed a dose-dependent improvement tendency in high-fat diet (HFD)-induced obesity, non-alcoholic fatty liver disease (NAFLD), hyperlipidemia, and diabetic nephropathy compared to the metformin (250 mg/kg). In particular, BCS extract at a dose of 200 mg/kg significantly inhibited the HFD-induced metabolic conditions. The oral administration of BCS extract (200 mg/kg) significantly inhibited the oxidative stress through lipid peroxidation, normalized the activity of sugar metabolism-related enzymes and the expression of genes involved in fat metabolism, and inhibited insulin resistance through glucose and fat metabolism by regulating the 5'-AMP-activated protein kinase (AMPK) expression. Furthermore, BCS extract (200 mg/kg) showed renal damage improvement effects compared to the metformin (250 mg/kg). The results clearly show that BCS aqueous extract at an appropriate concentration could help in the treatment of metabolic disorders, and BCS aqueous extract can be used as a functional food for various diabetic complications, such as obesity, diabetes, and NAFLD.

Preventive and Therapeutic Effects of Krill Oil on Obesity and Obesity-Induced Metabolic Syndromes in High-Fat Diet-Fed Mice.

Obesity increases the risks of metabolic syndromes including nonalcoholic fatty liver disease (NAFLD), diabetic dyslipidemia, and chronic kidney disease. Dietary krill oil (KO) has shown antioxidant and anti-inflammatory properties, thereby being a therapeutic potential for obesity-induced metabolic syndromes. Thus, the effects of KO on lipid metabolic alteration were examined in a high-fat diet (HFD)-fed mice model. The HFD model (n = 10 per group) received an oral gavage with distilled water as a control, metformin at 250 mg/kg, and KO at 400, 200, and 100 mg/kg for 12 weeks. The HFD-induced weight gain and fat deposition were significantly reduced in the KO treatments compared with the control. Blood levels were lower in parameters for NAFLD (e.g., alanine aminotransferase, and triglyceride), type 2 diabetes (e.g., glucose and insulin), and renal dysfunction (e.g., blood urea nitrogen and creatinine) by the KO treatments. The KO inhibited lipid synthesis through the modification of gene expressions in the liver and adipose tissues and adipokine-mediated pathways. Furthermore, KO showed hepatic antioxidant activities and glucose lowering effects. Histopathological analyses revealed that the KO ameliorated the hepatic steatosis, pancreatic endocrine/exocrine alteration, adipose tissue hypertrophy, and renal steatosis. These analyses suggest that KO may be promising for inhibiting obesity and metabolic syndromes.

A pilot study on the effect of formulation and individual muscle mass on vitamin C absorption in randomized clinical study.

The current study investigated that the vitamin C absorption in plasma depends on the individual muscle mass and the formulation including drinks (Vita 500), capsules, and tablets by using a randomized and double-blind clinical study. The volunteers were divided into two groups that depended on their muscle mass, including those whose muscle mass was greater than 40% ( ≥ $ \ge $ 40%) and less than 40% muscle mass (<40%). Levels of vitamin C in blood plasma was analyzed by high-performance liquid chromatography by ultraviolet detection (HPLC-UV). The existing HPLC method was modified according to lab conditions but maintained a constantly low pH sample reduction procedure. The analytical method validated stability, linearity, recovery, reliability, and accuracy. The vitamin C absorption was the highest at 120 min after ingesting Vita 500 (21.47 ± 15.99 µmol/L). It was higher in the group that has more than 40% muscle mass compared to other formulations, such as tablets and capsules. The results from the current study indicate that vitamin C formulations differently affect the vitamin C absorption, and its effect depends on the muscle mass. As the results, liquid type vitamin C formulations could enhance vitamin C absorption, which resulted in an improvement of vitamin C absorption according to muscle mass. PRACTICAL APPLICATION: The results of this study may recommend using vitamin C supplementation as liquid type. It may also provide evidence that people with higher muscle mass can absorb vitamin C more efficiently.

Curcumin Attenuates Sarcopenia in Chronic Forced Exercise Executed Aged Mice by Regulating Muscle Degradation and Protein Synthesis with Antioxidant and Anti-inflammatory Effects.

The aim of the current study is to investigate the effects of spray dry powders of Curcuma longa containing 40% curcumin (CM-SD), as a new aqueous curcumin formula, on sarcopenia in chronic forced exercise executed 10 month old ICR mice. CM-SD (80 and 40 mg/kg) increased calf thicknesses and strengths, total body and calf protein amounts, and muscle weights in both gastrocnemius and soleus muscles. mRNA expressions regarding muscle growth and protein synthesis were induced, while those of muscle degradation significantly declined in CM-SD treatment. CM-SD decreased serum biochemical markers, lipid peroxidation, and reactive oxygen species and increased endogenous antioxidants and enzyme activities. It also reduced immunoreactive myofibers for apoptosis and oxidative stress markers but increased ATPase in myofibers. These results suggest that CM-SD can be an adjunct therapy to exercise-based remedy that prevents muscle disorders including sarcopenia by anti-apoptosis, anti-inflammation, and antioxidation-mediated modulation of gene expressions related to muscle degradation and protein synthesis.

Curcumin Ameliorated Oxidative Stress and Inflammation-Related Muscle Disorders in C2C12 Myoblast Cells.

The purpose of the current study was to investigate antioxidant and anti-inflammatory effects of spray dry powder containing 40% curcumin (CM-SD) in C2C12 myoblast cells. CM-SD increased DPPH radical scavenging activity in a dose-dependent manner, and up to 30 µg/mL of CM-SD did not express cytotoxicity in C2C12 cells. Exposure to hydrogen peroxide (H2O2) drastically decreased the viability of C2C12 cells, but pre-treatment of CM-SD significantly increased the cell viability (p < 0.01). CM-SD significantly transactivated the nuclear factor erythroid-2-related factor 2 (Nrf2)-dependent luciferase activity in a dose-dependent manner and enhanced the levels of heme oxygenase (HO)-1, glutamate cysteine ligase catalytic subunit (GCLC), and NAD(P)H-dependent quinone oxidoreductase (NQO)-1. CM-SD also significantly reduced reactive oxygen species (ROS) production and lipid peroxidation and restored glutathione (GSH) depletion in H2O2-treated C2C12 cells. Moreover, CM-SD significantly reduced lipopolysaccharides (LPS)-mediated interleukin (IL)-6 production in the conditioned medium. Results from the current study suggest that CM-SD could be a useful candidate against oxidative stress and inflammation-related muscle disorders.

Physicochemical properties and bioavailability of naturally formulated fat-soluble vitamins extracted from agricultural products for complementary use for natural vitamin supplements.

The purpose of the current study was to evaluate the physicochemical properties, digestive stability, storage stability, and intestinal absorption of formulated natural vitamins (FNV) by mixing fat-soluble vitamins extracted from agricultural products with their synthetic vitamin (SYNV) counterparts using a 6 to 4 ratio (w:w, dry weight). The FNV A, D, E, and K were evenly dispersed without crystal growth in the dispersion specifications for the functional tablet foods. The FNV A, D, E, and K had 89, 73, 65, and 36% of the digestive recovery, respectively, which was comparable to that of the SYNV. FNV D, E, and K were retained over 77%, but rapidly decreased to 15% after 6 months during accelerated storage at 25 30 and 35℃. The comparable radical scavenging capacity was found between the FNV and the SYNV. Results from the current study suggest that fat-soluble vitamins extracted from agricultural products could be reasonable complementary use for natural vitamin supplements.

Ginseng berry aqueous extract prevents scopolamine-induced memory impairment in mice.

Ginseng berry exhibits a diverse range of pharmacological activities. The present study aimed to examine the neuroprotective effects of ginseng berry aqueous extract (GBE) against oxidative stress and to assess the impact of GBE on memory impairment in mice. In HT-22 cells, GBE pretreatment significantly inhibited glutamate- and hydrogen peroxide-mediated cytotoxicity in a concentration-dependent manner, while treatment with up to 100 µg/ml GBE alone did not change cell viability. In a murine model of scopolamine (SCP)-induced memory impairment, results from the passive avoidance test and the Morris water maze test indicated that GBE administration for 4 weeks prolonged step-through latency time and shortened escape latency time, suggesting that GBE can attenuate deficits in long-term memory induced by SCP. Additionally, GBE prevented SCP-induced reductions in acetylcholine by decreasing acetylcholinesterase activity and upregulating choline acetyltransferase mRNA levels in the hippocampus. GBE mitigated SCP-mediated mRNA decreases in brain-derived neurotrophic factor levels and its associated signaling molecules. Furthermore, GBE administration significantly suppressed malondialdehyde production and increased glutathione levels, catalase activity and superoxide dismutase activity in SCP-induced memory impaired mice. Therefore, the results of the current study indicated that ginseng berry may be a potential candidate for treating or preventing memory deficits that are associated with neurodegenerative disorders.

Hepatoprotective and anti-obesity effects of Korean blue honeysuckle extracts in high fat diet-fed mice.

PURPOSE: This study aimed to study the protective effects and mechanism of Blue Honeysuckle (BH) extracts (Berries of Lonicera caerulea L.) on non-alcoholic fatty liver disease (NAFLD) and obesity risk factors in a high fat-diet (HFD) model. METHODS: Animals adapted to HFD were selected after 1 week of adaption period and divided into 6 groups (8 mice in each group; 40 HFD-fed mice and 8 normal fat pellet diet (NFD)-fed mice). After the end of 12 weeks of continuous oral administrations of 3 different dosages of BH extract, 400, 200 and 100 mg/kg, or metformin 250 mg/kg, dissolved in a volume of 10 mL/kg distilled water, the hepatoprotective, hypolipidemic, hypoglycemic, nephroprotective, and anti-obesity effects were analyzed. RESULTS: The BH extract improved fat density and mass, adipocyte histopathology, hepatocyte hypertrophy, hepatic enzyme activity, lipid metabolism, and related gene expression including ACC1, AMPK α1 and AMPK α2 in hepatic tissue, leptin, UCP2, adiponectin, C/EBP α, C/EBPß and SREBP1c in adipose tissue. Especially, 200 mg/kg of BH extract constantly improved NAFLD and obesity risk factors through AMPK upregulation-mediated hepatic glucose enzyme activity, lipid metabolism-related gene expression, and activation of the antioxidant defense system, to a level comparable to that of metformin 250 mg/kg in HFD-fed mice. CONCLUSION: BH extract has the potential to reduce the risk factors associated with obesity, in addition to the remarkable effect of preventing NAFLD. Future research will need to be done to determine whether these results are consistent in human studies.