Anti-adipogenic Activity Research Articles

Japanese Leaf Burdock Extract Inhibits Adipocyte Differentiation in 3T3-L1 Cells.

Burdock, Arctium lappa Linn. (Asteraceae), is cultivated in East Asian for its edible roots, and its seeds are used in a herbal medicine. Burdock seeds and roots exhibit anti-adipogenic activity. However, the anti-adipogenic activity of the aboveground parts of burdock remains underexplored. Therefore, this study determined the anti-adipogenic effects of burdock leaf extract on 3T3-L1 adipocytes. Seventy percent ethanol (EtOH) extract of burdock leaves, which inhibited lipid accumulation, was fractionated into five fractions using Diaion® HP-20. EtOH eluted fractions (40 and 80%) strongly inhibited lipid accumulation. A common compound in these fractions was onopordopicrin (OPP), and purified OPP suppressed lipid accumulation and inhibited adipocyte differentiation. α, β-unsaturated carbonyl structure of OPP suggests potential electrophilic reactivity with polyfunctional thiol-trapping agents like cysteine residues. Indeed, the anti-adipogenic effects of OPP disappeared with the addition of cysteamine, which possesses a thiol group. Rhodamine-maleimide assay showed that OPP binds to the cysteine residues of adipocyte-specific transcription factor, peroxisome proliferator-activated receptor γ (PPARγ). Hence, our findings suggest that OPP exerts its anti-adipogenic action via binding to cysteine residues in signal proteins like PPARγ, inhibiting their activation. Thus, burdock leaf extract is a potential source of OPP, a bioactive compound with anti-obesity activity.

Bolanthus turcicus: a promising antidiabetic with in-vitro antioxidant, enzyme inhibitory and antiadipogenic activities.

It is crucial to investigate new anti-diabetic agents and therapeutic approaches targeting molecules in potential signaling pathways for the treatment of Type 2 diabetes mellitus (T2DM). The objective of the study was to investigate the total phenolic content, antioxidant capacity, α-glucosidase, and α-amylase inhibitory activities of Bolanthus turcicus (B. turcicus), as well as their cytotoxic, anti-adipogenic, anti-diabetic, apoptotic, and anti-migration potential on adipocytes. B. turcicus samples were extracted with methanol (MeOH), ethyl acetate (EA) and aqueous (Aq) solvents. The MeOH extract had the highest phenolic content (81.14mg GAE/g), followed by EA (74.93mg GAE/g) and Aq (51.09mg GAE/g). All extracts exhibited dose-dependent increases in α-glycosidase and α-amylase inhibitory activity. B. turcicus extracts showed cytotoxic effect on adipocytes with IC50 values of MeOH (141.0µg/mL) < Aq (155.3µg/mL) < EA (199.5µg/mL). Furthermore, B. turcicus extracts reduced lipid droplet formation and adipocyte diameter size. All extracts altered cell morphology to resemble fibroblasts. B. turcicus extracts exhibited anti-migratory effect delaying wound healing for up to 96h. The B. turcicus extracts showed a pro-apoptotic effects on adipocytes by increasing Caspase-3 enzyme activity and the population of DAPI-positive cell with apoptotic nuclear-morphology. B. turcicus extracts upregulated the expression of the Glut-4 gene at the mRNA, protein and intracellular level in adipocytes. In conclusion, our findings indicate that B. turcicus not only exhibits strong antioxidant properties and enzyme inhibitory activities but also exerts significant anti-adipogenic and pro-apoptotic effects in adipocytes, thereby providing a comprehensive mechanism through which it may contribute to the management of T2DM. These effects highlight the potential of B. turcicus as a therapeutic agent for improving glucose homeostasis and insulin sensitivity.

Evaluation of Functional Properties and the Lipase Inhibitory Activity of Proteins from Amaranthus cruentus Seeds

Aims/Background: Obesity is characterized by excessive body fat and is a chronic and complex disease. The medications used to date for the treatment of obesity have exhibited various side effects. Thus, new substances must be sought from alternative sources as anti-obesity drugs. Recently, there has been increased interest in the study of bioactive peptides generated from plant proteins for functional food development. Plant proteins have exhibited pancreatic lipase inhibitory activity and can be potentially studied as sources of anti-obesity drugs for long-term use. Amaranthus cruentus seeds are very nutritious, with high protein con-tent and medicinal properties. Hence, this study focuses on pancreatic lipase inhibitory activity and functional properties of Amaranthus cruentus seed protein isolates. Materials and Methods: Proteins were isolated using conventional and thermal isoelectric pre-cipitation methods. The protein's ability to inhibit lipases was assessed using both synthetic and natural substrates. The functional properties, such as nitrogen solubility, foaming test, emulsi-fication test, and water and oil holding abilities, were evaluated. The protein isolates were char-acterized by SDS-PAGE and DSC. Cell line studies were performed on 3T3-L1 cell lines to determine the cytotoxicity and anti-adipogenic activity of protein isolates. Results: Proteins 1, 2, 3, and 4 were found to have high protein yield and protein content. SDS-PAGE characterization showed protein bands at different molecular weights (kDa). Protein iso-lates demonstrated good functional properties and antilipase activity as compared to flour. In the cell line experiments, protein 1 exhibited a dose-dependent reduction in the lipid content. Conclusion: A. cruentus seed protein could pave the way for the development of nutraceutical formulation in the treatment of obesity.

Anti-adipogenic activity of oleoresin from Nigella sativa L. seeds via modulation of PPAR-γ and C/EBP-α expression in 3T3-L1 adipocytes

Anti-adipogenic activity of oleoresin from Nigella sativa L. seeds via modulation of PPAR-γ and C/EBP-α expression in 3T3-L1 adipocytes

Cell-penetrating anti-sense peptide nucleic acids targeting sulfatase 2 inhibit adipogenesis in human mesenchymal stem cells

Targeting the genes regulate the lineage commitment of human mesenchymal stem cells (hMSCs) to adipocytes provides a promising strategy for addressing obesity. In this study, we investigated the therapeutic potential of cell-penetrating anti-sense peptide nucleic acids (PNAs) designed to enhance solubility and hybridization properties, specifically targeting sulfatase 2 (SULF2), a potential reciprocal regulator of adipocyte and osteoblast differentiation in hMSCs. Cell-penetrating modified PNA oligomers effectively inhibit SULF2 gene transcription, leading to significant reductions in adiponectin protein synthesis and intracellular lipid droplet accumulation during adipogenesis in human bone marrow-derived MSCs (hBM-MSCs). Notably, PNA oligomer compound 5 exhibited the most potent anti-adipogenic activity, with an IC50 value of 0.28 μM. These findings show the potential of SULF2-targeting cell-penetrating PNA oligomers as novel therapeutic agents for obesity-related metabolic diseases.

GC-MS SCRUTINY OF PHYTOCONSTITUENTS, IN VITRO EVALUATION OF ANTIHYPERGLYCEMIC, ANTIADIPOGENIC ACTIVITIES, AND CYTOTOXIC EFFECT USING 3T3 L1 ADIPOCYTE CELL LINE AND MOLECULAR DOCKING STUDIES OF PREMNA CORYMBOSA

Objective: The present study was intended to list out the phytochemical multiples and to investigate the antihyperglycemic effect of Premna corymbosa using in vitro assays and in silico molecular docking methods. Methods: The phytochemical multiples of methanol proportion of P. corymbosa leaves were appraised by Gas Chromatography-Mass Spectrometry (GC-MS) scrutiny to illustrate the attendance of phytochemical composites. Moreover, the in vitro antihyperglycemic, antiadipogenic activities, and cytotoxic effects of the extract were elucidated using a 3T3 L1 adipocyte cell line. Mode of action of phytochemical composites in methanol leaf extract of P. corymbosa was probed by Western blotting with IRS1, IRS2, mTOR, and glucose transporter type 4 (GLUT 4) receptors. At present, to probe the consequence of the aboriginal drugs, it is necessary to perform in silico docking on the diabetic receptor which could be useful for the progress of enhanced formulation for the psychoanalysis of diabetes. Results: The GC-MS scrutiny depicted the being there of thirty-five phytochemical multipart. Amid the thirty-five multipart’s recognized, focal composites were Phytol, acetate (RT-16.78), n-Hexadecanoic acid (RT-18.16), Phytol (RT-19.51), 9,12,15-octadecatrienoic acid (Z,Z,Z) (RT-19.85), octadecanoic acid (RT-20.04), and Bis (2-ethylhexyl) phthalate (RT-23.09). The results of the glucose conception assay, adipocyte differentiation assay, and MTT assay showed potent in vitro antihyperglycemic activity with methanol leaf extract of P. corymbosa in 3T3l1Cell line. The results attained from western blotting revealed good antihyperglycemic activity of P. corymbosa. The in silico molecular docking results illustrated that the selected herbal lead compound is an effective target against the receptors. The compound showed favorable interactions with the amino acid residues thereby substantiating their proven efficacy as an antihyperglycemic compound. Conclusion: The outcome of the current study substantiates the antihyperglycemic prospective of the methanol leaf extract of P. corymbosa on the hyperglycemic causal agents and its activity against diabetes by a molecular approach.

Identification of Bioactive Substances Derived from the Probiotic-Induced Bioconversion of Lagerstroemia speciosa Pers. Leaf Extract That Have Beneficial Effects on Diabetes and Obesity.

Lagerstroemia speciosa L. (Banaba) has been used as a functional food because of its diuretic, decongestant, antipyretic, anti-hyperglycemic, and anti-adipogenic activities. Triterpene acids, including corosolic acid, oleanolic acid, and asiatic acid, are the principal phytochemicals in Banaba and are potentially anti-diabetic substances, owing to their effect on blood glucose concentration. Bioconversion of Banaba leaf extract (BLE) by Lactobacillus plantarum CBT-LP3 improved the glucose uptake, insulin secretion, and fat browning of this functional food. Furthermore, we identified asiatic acid, which was found to be increased by 3.8-fold during the L. plantarum CBT-LP3-mediated bioconversion process using metabolite profiling. Most previous studies have focused on corosolic acid, another triterpene acid that is a known anti-diabetic compound and is used to standardize BLE preparations. However, asiatic acid is the second most common of the triterpene acids and is also well known to have anti-diabetic properties. The present study has provided strong evidence that asiatic acid represents an alternative to corosolic acid as the most important active compound. These results suggest that the probiotic-mediated bioconversion of BLE may improve the anti-diabetic effects of this functional food. This implies that the consumption of a probiotic should be encouraged for people undergoing BLE treatment to improve its anti-diabetic effects.

Advanced Nanotechnological Approaches for Effective Delivery of Rutin: An Updated Review.

Polyphenols are a group of naturally occurring compounds that have intriguing biological activities. Among these compounds is rutin, a polyphenolic flavanol found in many plants, including passion flowers, buckwheat seed, fruits and fruit rinds, and citrus fruits (such as orange, grapefruit, lemon, and lime). Various studies have demonstrated rutin to possess antibacterial, antifungal, antiallergic, anti-inflammatory, anti-diabetic, anti-adipogenic, anti-carcinogenic, anti-apoptotic, anti-osteoporotic, radioprotective, gastroprotective, neuroprotective, and nephroprotective activities. Despite its benefits, rutin's therapeutic applicability is severely limited due to its low water solubility, sensitivity to oxidation, and dissolving rate. However, these problems can be overcome by employing an efficient delivery approach. An extensive number of nanocarriers can be developed for medicinal use if pre-clinical as well as human-clinical studies are completed. The current review presents an overview of effective rutin nano-formulations for targeted therapy in various health disorders. This review article discusses the clinical evidence, current status, as well as future opportunities of rutin nanocarriers for increasing rutin's bioactivity for possible medicinal uses.

Therapeutic potential of ethoxy mansonone G: A comprehensive exploration of its anticancer actions in breast cancer, colorectal cancer, and non-small cell lung carcinoma.

Mansonone G (MG), a 1,2-naphthoquinones with antiestrogenic, antimicrobial, and anti-adipogenic activities, is derived from the heartwood of Mansonia gagei Drumm. Ethoxy mansonone G (EMG), an essential derivative of MG, has anticancer and antioxidant agent. EMG also has antiestrogen activity and is demonstrated to lower estrogen receptorexpression in endocrine-resistant cells. EMG significantly inhibits cell division, invasion, and anchorage-dependent growth in all cancer types. Through the stimulation of the tumor protein (p53) and extracellular signal-regulated kinase (ERK) signaling cascades, it also causes apoptosis. Moreover, it manifests its anti-cancerous effects in toll-like receptor pathways, c-Jun N-terminal kinase (c-JNK), and nuclear factor kappa B (NF-κB). EMG inhibits the phosphorylation of glycogen synthase kinase (GSK3), Erk, protein kinase B (Akt), and mammalian target of rapamycin (mTOR). By interfering with molecular cascades, EMG significantly reduces the metabolism of cancer cells. This paper focuses on the potential use of EMG in cancer treatment. Moreover, it states the methodology by which specific assays establish the anti-cancerous role of EMG. Breast cancer, non-small cell lung cancer, and colorectal cancer are only a few of the cancers for which EMG was shown to be effective. Through further research, EMG may be developed as a therapeutic solution to complications caused by cancer. This study presents EMG as a novel candidate for cancer therapy, offering a unique combination of pharmacological advantages and mechanistic insights that warrant further exploration and development toward addressing the complexities of cancer treatment.

Anti-Obesity Activities of the Compounds from Perilla frutescens var. acuta and Chemical Profiling of the Extract.

Perilla frutescens var. acuta (Lamiaceae) is widely used not only as an oil or a spice, but also as a traditional medicine to treat colds, coughs, fever, and indigestion. As an ongoing effort, luteolin-7-O-diglucuronide (1), apigenin-7-O-diglucuronide (2), and rosmarinic acid (3) isolated from P. frutescens var. acuta were investigated for their anti-adipogenic and thermogenic activities in 3T3-L1 cells. Compound 1 exhibited a strong inhibition against adipocyte differentiation by suppressing the expression of Pparg and Cebpa over 52.0% and 45.0%, respectively. Moreover, 2 inhibited the expression of those genes in a dose-dependent manner [Pparg: 41.7% (5 µM), 62.0% (10 µM), and 81.6% (50 µM); Cebpa: 13.8% (5 µM), 18.4% (10 µM), and 37.2% (50 µM)]. On the other hand, the P. frutescens var. acuta water extract showed moderate thermogenic activities. Compounds 1 and 3 also induced thermogenesis in a dose-dependent manner by stimulating the mRNA expressions of Ucp1, Pgc1a, and Prdm16. Moreover, an LC-MS/MS chromatogram of the extract was acquired using UHPLC-MS2 and it was analyzed by feature-based molecular networking (FBMN) and the Progenesis QI software (version 3.0). The chemical profiling of the extract demonstrated that flavonoids and their glycoside derivatives, including those isolated earlier as well as rosmarinic acid, are present in P. frutescens var. acuta.

Antiadipogenic activity of homoeopathic preparation of Chelidonium Majus employing3T3-L1 cell line as a model.

Adipocytes are derived from mesenchymal stem cells through the process of adipogenesis. Adipocyte metabolism is abnormal in a range of disorders like obesity, nutritional insufficiency and diabetes. The 3T3-L1 preadipocytes are most often used to create adipocyte models, and they can be differentiated into adipocyte cells under the right conditions. Homeopathic preparation of Chelidonium majus (HPCM) in management of Obesity and T2DM has many evidences in clinical practice.3T3-L1 preadipocyte cells were differentiated into adipocytes by using differentiation cocktail. Cell were treated with HPCM attenuation at the concentration of 0.5%, 1% and 2% for 15 days. Oil O Red Staining was used to assess LD accumulation. In order to determine the lipid content in 3T3-L1 adipocytes, cells were dissolved in isopropanol and the absorbance values were measured. Images were captured and analysed using software imageJ.We investigated the action of Homeopathic preparations of Chelidonium majus (HPCM) in obesity using the 3T3-L1 adipogenesis model employing huMSC’s.After differentiation of adipocytes many LDs were formed in 3T3-L1 preadipocytes which can be compared with no lipid droplet in 3T3-L1nondifferentiated cells. Further, area of differentiated adipocytes was mapped and then compared for LD accumulation in control and HPCM for checking its antiadipogenic activity.A significant reduction in accumulation of lipid droplets was seen in 0.5%, 1% and 2% concentration of HPCM as compared to control during the differentiation of 3T3-L1 preadipocytes into adipocytes. Excessive differentiation of cells and high fat accumulation in the adipose tissue are closely linked to obesity. Preadipocyte differentiation inhibitors may have preventive and therapeutic potential as anti-obesity drugs. HPCM has potential to act as an antiadipogenic agent which can be used to combat various diseases like obesity, T2DM and cardiovascular diseases

S-Dihydrodaidzein and 3-(1,3-benzoxazol-2-yl)-benzamide, Two New Potential β-estrogen Receptor Ligands with Anti-adipogenic Activity.

The elucidation of molecular pathways associated with adipogenesis has evidenced the relevance of estrogen and estrogen receptor beta (ERβ). The positive effects of ERβ ligands on adipogenesis, energy expenditure, lipolysis, food intake, and weight loss, make ERβ an attractive target for obesity control. From ligand-based virtual screening, molecular docking, and molecular dynamic simulations, six new likely ERβ ligands (C1 to C6) have been reported with potential for pharmacological obesity treatment. In this study, the effect of molecules C1-C6 on adipogenesis using the murine 3T3-L1 cell line was evaluated. Cell viability was assessed by MTT assays. Lipid accumulation and gene expression were investigated by ORO staining and real-time quantitative RT-PCR experiments, respectively. Cell viability was not significantly affected by C1-C6 at concentrations up to 10 μM. Interestingly, treatment with 10 μM of C1 (S-Dihydrodaidzein) and C2 (3-(1,3-benzoxazol-2-yl)- benzamide) for 72 h inhibited adipocyte differentiation; moreover, ORO staining evidenced a reduced intracellular lipid accumulation (40% at day 7). Consistently, mRNA expression of the adipogenic markers, PPARγ and C/EBPα, was reduced by 50% and 82%, respectively, in the case of C1, and by 83% and 59%, in the case of C2. Altogether, these results show the two new potential β-estrogen receptor ligands, C1 and C2, to exhibit anti-adipogenic activity. They could further be used as lead structures for the development of more efficient drugs for obesity control.

Identification of novel pharmacological activators of circadian clock with anti-adipogenic properties

The circadian clock drives rhythmic oscillations of metabolic processes to orchestrate metabolic homeostasis, and disruption of this mechanism predisposes to obesity and insulin resistance. Preserving or augmenting clock function could be a potential therapeutic target for metabolic diseases, particularly with the wide-spread of circadian misalignment in a modern lifestyle. To date, targeting the clock for metabolic disease prevention or treatment remains to be explored. Adipocyte possesses cell-autonomous clocks that exert transcriptional control of the Wnt pathway to inhibit adipocyte development. Using a high through-put screening pipeline, we recently identified Chlorhexidine as a novel clock-activating molecule that targets the key driver of the circadian clock transcriptional feedback loop, the CLOCK protein. Based on clock function in suppressing adipogenesis, we hypothesize that Chlorhexidine and related compounds may possess adipogenic-inhibitory properties suitable for anti-obesity drug development. In distinct adipogenic progenitor models, we demonstrate the activity of Chlorhexidine in inhibiting adipogenic lineage commitment and terminal differentiation. Furthermore, we report the structural optimization of Chlorhexidine chemical scaffold that led to the discovery of new analogs with improved anti-adipogenic effcacy. Consistent with its activity for clock activation, in adipogenic progenitors containing a Period2::dLuc luciferase reporter, Chlorhexidine induced significant shortening of clock period length with induction of core clock components. Chlorhexidine treatment of adipogenic mesenchymal precursors robustly suppressed their adipogenic maturation, with a comparable effect observed on inhibiting terminal differentiation of primary preadipocytes in a clock-dependent manner. Mechanistically, Chlorhexidine stimulates clock-controlled Wnt signaling that mediates its anti-adipogenic effect. Through medicinal chemistry modification of its chemical scaffold, we generated a panel of Chlorhexidine analogs with validation for their clock-modulatory activities. Structure activity relationship analysis of these analogs led to the identification of CM002 as a new clock activator with improved clock-dependent anti-adipogenic activity. Collectively, our findings uncovered a new class of clock-modulatory compounds that inhibit adipocyte development for potential anti-obesity drug discovery and provide clock-targeting chemical probes for dissecting clock function in metabolic physiology. This project was supported by grants from National Institute of Aging R56AG080294 and Arthur Riggs-Diabetes &amp; Metabolism Research Institute T2D and T1D Innovative Awards to KM. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Structurally Diverse Bisbenzylisoquinoline Alkaloids with Antiadipogenic Activity through PPARγ Downregulation from the Embryo of Nelumbo nucifera Seeds.

Six undescribed and six known bisbenzylisoquinoline alkaloids were isolated from the embryo of Nelumbo nucifera seeds. Their structures were fully characterized by a combination of 1H, 13C NMR, 2D NMR, and HRESIMS analyses, as well as ECD computational calculations. The antiadipogenic activity of 11 alkaloids was observed in a dose-responsive manner, leading to the suppression of lipid accumulation in 3T3-L1 cells. Luciferase assay and Western blot analysis showed that the active alkaloids downregulated peroxisome proliferator-activated receptor gamma (PPARγ, a key antiadipogenic receptor) expression in 3T3-L1 cells. Analysis of the structure-activity relationship unveiled that a 1R,1'S configuration in bisbenzylisoquinoline alkaloids led to a notable enhancement in antiadipogenic activity. The resistance level against lipid accumulation highlighted a consistent pattern with the suppressive effect on the PPARγ expression. These activity results indicate that alkaloids from the embryo of N. nucifera seeds have a potential of antiobesity effects through PPARγ downregulation.

Facile adipocyte uptake and liver/adipose tissue delivery of conjugated linoleic acid-loaded tocol nanocarriers for a synergistic anti-adipogenesis effect

Obesity is a major risk to human health. Adipogenesis is blocked by α-tocopherol and conjugated linoleic acid (CLA). However, their effect at preventing obesity is uncertain. The effectiveness of the bioactive agents is associated with their delivery method. Herein, we designed CLA-loaded tocol nanostructured lipid carriers (NLCs) for enhancing the anti-adipogenic activity of α-tocopherol and CLA. Adipogenesis inhibition by the nanocarriers was examined using an in vitro adipocyte model and an in vivo rat model fed a high fat diet (HFD). The targeting of the tocol NLCs into adipocytes and adipose tissues were also investigated. A synergistic anti-adipogenesis effect was observed for the combination of free α-tocopherol and CLA. Nanoparticles with different amounts of solid lipid were developed with an average size of 121‒151 nm. The NLCs with the smallest size (121 nm) showed greater adipocyte internalization and differentiation prevention than the larger size. The small-sized NLCs promoted CLA delivery into adipocytes by 5.5-fold as compared to free control. The nanocarriers reduced fat accumulation in adipocytes by counteracting the expression of the adipogenic transcription factors peroxisome proliferator activated receptor (PPAR)γ and CCAAT/enhancer-binding protein (C/EBP)α, and lipogenic enzymes acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS). Localized administration of CLA-loaded tocol NLCs significantly reduced body weight, total cholesterol, and liver damage indicators in obese rats. The biodistribution study demonstrated that the nanoparticles mainly accumulated in liver and adipose tissues. The NLCs decreased adipocyte hypertrophy and cytokine overexpression in the groin and epididymis to a greater degree than the combination of free α-tocopherol and CLA. In conclusion, the lipid-based nanocarriers were verified to inhibit adipogenesis in an efficient and safe way.

Ginseng (Panax ginseng) leaf extract modulates the expression of heme oxygenase-1 to attenuate osteoclast differentiation

Osteoporosis is an aging disease characterized by an imbalance between bone formation and resorption. However, drugs that inhibit bone resorption have various adverse effects. Ginseng (Panax ginseng), a prominent herbal medicine in East Asia for >2000 years, is renowned for its manifold beneficial properties, including antioxidant, anti-cancer, anti-diabetic, and anti-adipogenic activities. Despite its long history of use, the pharmacological functions of ginseng leaves are not yet fully comprehended. In this study, we evaluated the potential effects of ginseng leaf extract (GLE) on receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclast differentiation in RAW264.7 macrophage cells. Tartrate-resistant acid phosphatase (TRAP) staining revealed that GLE had significant anti-osteoclastogenic activity. GLE significantly reduced mRNA levels of osteoclast differentiation markers including TRAP, nuclear factor of activated T cell cytoplasmic 1, and cathepsin K. It also suppressed the production of reactive oxygen species (ROS) and secretion of high mobility group box-1 (HMGB1) in RANKL-treated RAW264.7 cells. In addition, GLE upregulated dose- and time-dependently the expression of heme oxygenase-1 (HO-1), eventually suppressing ROS production and HMGB1 secretion. This effects of GLE were significantly reversed by Tin Protoporphyrin IX dichloride, an inhibitor of HO-1, and HO-1 shRNA, indicating that HO-1 potently inhibits RANKL-induced osteoclast differentiation by inhibiting ROS production and HMGB1 secretion. Taken together, these observations suggest that GLE could have therapeutic potential as a natural product-derived medicine for the treatment of bone disorders.

Cytotoxicity, Antiadipogenic, Low‐Density Lipoprotein Oxidation Inhibitory Activities, and Acute Toxicity Study of Psychotria densinervia Hydroethanolic Leaf and Bark Extracts

Background: Obesity is increasingly taking an important stage as a cause of death worldwide, and interventions with a good cost‐effectiveness ratio are needed. Psychotria densinervia is one of these natural products with health benefits. Objective. The present study evaluated the cytotoxicity, antiadipogenic, low‐density lipoprotein (LDL), oxidation inhibitory activities, and acute toxicity of Psychotria densinervia hydroethanolic leaf and bark extracts.Methods: The cytotoxicity evaluation of the extracts (62.5, 125, 250, and 500 μg/mL) using the MTT assay and the antiadipogenic activity (25, 50, 100, and 200 μg/mL) using oleic acid were carried out in SW‐872 cells. Copper sulfate (CuSO4)‐induced oxidation was used in the evaluation of the effect of extracts (0.25, 0.5, and 1 mg/mL) against LDL oxidation. The oral acute toxicity evaluation of a single dose of 2000 mg/kg of the extracts was performed in Wistar albino rats weighing 127 ± 2 g.Results: The leaf and bark extracts did not show any sign of cytotoxicity at the tested concentrations. The best antiadipogenic activity was observed by the standard orlistat (38.45 ± 1.70 μg/mL), followed by the leaf extract (IC50: 41.47 ± 0.50 μg/mL) and the least the bark extract (IC50: 107.50 ± 0.90 μg/mL). At a concentration of 1 mg/mL, the leaf extract presented an oxidation lag time of 130 min, which was higher and better than that of the bark extract (120 min). Quercetin (standard) presented an oxidation lag time longer than 3 h. The oral acute toxicity evaluation did not show any signs of toxicity indicating that the LD50 was greater than 2000 mg/kg.Conclusion: Based on the results obtained, the P. densinervia hydroethanolic leaf extract possesses a better antioxidant and antiadipogenic activities than the bark extract.

Toxicological Evaluation of a Polyherbal Formulation (18KHT01) and Validation of UPLC-DAD Method for Quality Control.

Background: 18KHT01 is a novel synergistic composition of Quercus acutissima, Camellia sinensis, Geranium thunbergii, and a small portion of Citrus limon. Our previous report demonstrated that the 18KHT01 exhibits potent antiobesity effects, with synergistic antioxidant, antiadipogenic, and antiobesity activities in diet-induced obese mice. This study explores the toxicity profile and quality control parameters of the 18KHT01 formulation. Methods: Broad-spectrum acute and subacute oral toxicity studies were performed using male and female ICR mice. In order to simultaneous analysis of the 18KHT01 formulation, an ultraperformance liquid chromatography coupled with a diode array detector (UPLC-DAD) method was developed and validated using six marker compounds. Results: Acute oral toxicity evaluation of 18KHT01, administered at single high doses of 2, 2.5, 3, and 5 g/kg, identified 2 g/kg as the no-observed adverse effect level (NOAEL). The LD50 (50% lethal dose) and LD100 (100% lethal dose) of 18KHT01 for male ICR mice were 3.99 and 7.77 g/kg, and those for the female mice were 2.94 and 4.70 g/kg, respectively. In addition, a 30-day repeated dose oral subacute toxicity evaluation indicated that 18KHT01 is safe below 500 mg/kg/day for long-term administration in ICR mice of either sex. UPLC-DAD method validation revealed that each calibration curve for the marker compounds showed good linearity, as well as the validation parameters such as precision, specificity, and accuracy met the acceptance criteria. Conclusion: The present study evidenced the toxicological profile of 18KHT01 polyherbal formulation in mice as well as developed a simple, rapid, and accurate chromatographic method for quality control.

Micro-grinding-based production for sulforaphene-enriched radish seeds extract via facilitating glucosinolates-myrosinase reaction, and evaluation of its anti-adipogenic effects.

Sulforaphene (SFEN), an isothiocyanate (ITC) abundant in radish (Raphanus sativus) seeds (RS), has many health benefits, including anti-obesity effects. SFEN content is affected by multiple factors during processing, such as glucoraphenin (GLE) (the precursor of SFEN) availability, myrosinase (essential for conversion from GLE to SFEN) activity, and SFEN stability. We examined the physiochemical-properties and anti-adipogenic effects of SFEN-enriched RSE produced by two processes, roasting and micro-grinding. The roasting process lowered SFEN content and myrosinase activity over 50°C. However, among micro-grinding conditions, smaller particle size (#2 grind, ≈11.31μm) more effectively increased SFEN content in RS compared to larger particles (#1 grind, ≈ 179.50 μm) by accelerating available GLE and myrosinase release from RS. Grind #2 also effectively inhibited the adipogenesis of 3T3-L1 pre-adipocytes compared to #1. Thus, micro-grinding can be suggested for producing SFEN-enriched RSE with anti-adipogenic activity as a functional material for obesity prevention or treatment.

A quality evaluation method of lotus leaf based on its lipid lowering components using QAMS and chemometrics

IntroductionLotus leaf has long been used as food and medicine in China and is well-known for its lipid-lowering effects. However, there is a lack of a comprehensive quality evaluation for lotus leaf due to the absence of consideration of the correlation between various components and their efficacy. ObjectivesThis study aims to find out the key bioactive components that can be used for quality evaluation of lotus leaf on lipid-lowering effect. MethodsThirteen compounds were characterized in the lotus leaf using ultra-high- performance liquid chromatography-time-of-fight mass spectrometry (UPLC-Q-TOF-MS). Five alkaloids and four flavonoids were identified according to their lipid-lowering activities reported in literatures. Then, the contents of these nine components were analyzed in 39 batches of lotus leaves growing in different locations using high performance liquid chromatography diode-array detector (HPLC-DAD), and further evaluated by quantitative analysis of multi-components by single marker (QAMS) and chemometrics. The anti-adipogenic activity of lotus leaves were evaluated for their inhibitory effect on the PPARγ expression by luciferase assay. ResultsThe 39 batches were clustered into two regions, the north and the south, based on the contents of these components. Three alkaloids, nuciferine, N-nornuciferine, and asimilobine, and three flavonoids, astragalin, hyperoside, and trifolioside, were found to serve as the key factors behind the region differences. Their contents were higher in Guangchang County of Jiangxi Province than other habitat locations. Moreover, the luciferase assay combined with chemometrics showed that these components were positively correlated with lipid-lowering activity of lotus leaf. ConclusionsThree alkaloids and three flavonoids were screened out and could be used as key compounds for quality evaluation of lotus leaf on lipid-lowering effect.