3T3-L1 Cell Line Research Articles

Isolation, characterization and anticancer activity of secondary metabolites from Verbascum speciosum.

Herein, two iridoid glucosides aucubin (1) and ajugol (2), and two phenyl ethanoids, verbascoside (3) and poliumoside (4) were isolated from the methanol extract of the aerial parts of Verbascum speciosum and used to study about their anticancer activity for the first time. The structures of all compounds were elucidated using spectroscopic data (IR, 1D and 2D NMR, LC-TOF/MS). Antiproliferative activities of Aucubun (1) and Verbascoside (3) were tested against A-549 (human colon cancer), MDA-MD-453 (human breast cancer) and 3T3-L1 (mouse fibroblast)cell lines by XTT assay. In addition, the anticarcer mechanism of action of aucubin (1) was investigated on MDA-MB-453 cells for the first time. XTT result showed that both applied compounds exhibited antiproliferative effect at different dose ranges depending on the cancer type, as well as selectivity between cancer and healty cell lines. Flow cytometry analyzes revealed that aucubin (1) exerts its cytotoxic effect in MDA-MB-453 cells by directing cells to early apoptosis and inhibiting the P13K/AKT signaling pathway.

Synergistic effect of Camellia sinensis and Withania somnifera on hydrogen peroxide-induced oxidative stress: an in vitro study

Camellia sinensis (CS) and Withania somnifera (WS) are well-known to have antioxidant and anti-ageing effects. However, information on their synergistic antioxidant effect is still not known. In this study, we have compared the % cell viability of oxidative stress induced 3T3-L1 cell lines after treatment with CS, WS alone and in combination. The methanolic extracts of CS and WS were prepared by three cycles of maceration with 24 h duration each. The effect of CS and WS alone and in combination was determined under three different conditions viz. pre-treatment, co-treatment and post-treatment at 24hrs, 48hrs and 72hrs with the help of MTT assay. CS+WS were found to show synergistic effect under pre-treatment conditions with % cell viability of 95.39 ± 0.7% at an optimum concentration of WS+CS (4.87μg/ml+15.22μg/ml) after 72 hrs. The combination index was found to be less than 1 at 24 hrs (0.278), 48 hrs (0.172) and 72 hrs (0.126) for combination of CS+WS indicating a synergistic effect of plant combination.

Long noncoding RNA ACART knockdown decreases 3T3-L1 preadipocyte proliferation and differentiation.

Obesity is a main risk factor for diabetes and cardiovascular disorders and is closely linked to preadipocyte differentiation or adipogenesis. Peroxisome proliferator-activated receptor γ (PPARγ) is an indispensable transcription factor in adipogenesis. A newly identified long noncoding RNA, Acart, exerts a protective effect against cardiomyocyte injury by transactivating PPARγ signaling. However, the function of Acart in preadipocyte differentiation is unclear. To investigate the function of Acart in adipogenesis, a well-established preadipocyte, the 3T3-L1 cell line, was induced to differentiate, and Acart level was assessed during differentiation using quantitative real-time PCR. The biological role of Acart in adipogenesis was analyzed by assessing lipid droplet accumulation, PPARγ and CCAAT/enhancer-binding protein α (C/EBPα) expression, and 3T3-L1 cell proliferation and apoptosis after Acart silencing. We found that Acart level was promptly increased during preadipocyte differentiation in vitro. Acart was also significantly upregulated in obese mouse-derived subcutaneous, perirenal, and epididymal fat tissues compared with nonobese mouse-derived adipose tissues. Functionally, Acart depletion inhibited preadipocyte differentiation, as evidenced by a significant decrease in lipid accumulation and PPARγ and C/EBPα expression levels. Acart silencing also inhibited 3T3-L1 cell proliferation, whereas Acart overexpression accelerated 3T3-L1 cell proliferation and decreased cell apoptosis. Taken together, the current results reveal a novel function of Acart in regulating preadipocyte proliferation and differentiation.

Novel Peptides in Fermented Milk with Specific Lactobacillus Strains Potential Antiobesity Effect: In Vitro and in Silico Analysis

The objective was to investigate the potential antiobesity effect of fermented milks (FMs) with 11 strains of specific Lactobacillus strains isolated from artisanal Mexican cheeses. First, pancreatic lipase inhibitory activity (PLIA) was determined in all FMs. Then, the FM with the most PLIA was selected for peptide identification by high-performance liquid chromatography (HPLC)/tandem mass spectrometry (MS–MS) and for adipogenesis studies. FMs with Limosilactobacillus fermentum J20 (J20) displayed the highest (p < 0.05) PLIA (81.70 ± 2.24%) and the lowest IC50 (0.05 0.05 mg/mL) of all. Also, FM J20 inhibited lipid accumulation in a 3T3-L1 cell line. Eight novel identified peptides released by in silico digestion from the fraction with the most PLIA (p < 0.05) presented sequences that strongly bind enzyme active sites and may be responsible for lipid accumulation inhibition during adipogenesis. Thus, FM J20 may be used for the development of a dairy product with potential antiobesity effects.

Sesamol promotes browning of white adipocytes through liver-adipose crosstalk signal of hepatic fibroblast growth factor 21

Sesamol (SEM), a lignan from sesame oil, exhibited potential benefits on obesity treatment by promoting browning of adipocytes, and the current study is aimed to explore the molecular mechanisms of SEM from the aspect of systemic liver-adipose crosstalk that mediated by hepatic fibroblast growth factor 21 (FGF21). Our in vivo data showed that SEM induced energy expenditure and white adipose tissue (WAT) browning by increasing the expression level of uncoupling protein-1 in high fat diet induced obese C57BL/6J mice. Elevated levels of circulating FGF21 associated with the increased expression of hepatic FGF21 were observed after SEM intervention. Simultaneously, the increased adipose fibroblast growth factor tyrosine kinase receptor 1/beta-klotho indicated that FGF21 sensitivity was enhanced by SEM in WAT. Furthermore, our in vitro results from HepG2 and 3T3-L1 cell lines confirmed the effects and revealed the mechanism of SEM on the white adipocytes browning. We found that with the specific inhibitors of PPARα, the SEM-mediated hepatic FGF21 expression was decreased, and with the specific inhibitors of PPARγ, the browning effect of adipocytes by SEM combining with FGF21 was significantly suppressed. Taken together, the mechanism of SEM for inducing the WAT browning might be the modulation of SEM on liver-adipose crosstalk mediated by FGF21, and the PPARs family might be the targets of SEM. The novel findings from the present study provided evidence that SEM could be a potent obesity-treating compound.

In Vitro Insulin Resistance Model: A Recent Update.

Insulin resistance, which affects insulin-sensitive tissues, including adipose tissues, skeletal muscle, and the liver, is the central pathophysiological mechanism underlying type 2 diabetes progression. Decreased glucose uptake in insulin-sensitive tissues disrupts insulin signaling pathways, particularly the PI3K/Akt pathway. An in vitro model is appropriate for studying the cellular and molecular mechanisms underlying insulin resistance because it is easy to maintain and the results can be easily reproduced. The application of cell-based models for exploring the pathogenesis of diabetes and insulin resistance as well as for developing drugs for these conditions is well known. However, a comprehensive review of in vitro insulin resistance models is lacking. Therefore, this review was conducted to provide a comprehensive overview and summary of the latest in vitro insulin resistance models, particularly 3T3-L1 (preadipocyte), C2C12 (skeletal muscle), and HepG2 (liver) cell lines induced with palmitic acid, high glucose, or chronic exposure to insulin.

Chronic Exposure to Low-Molecular-Weight Polycyclic Aromatic Hydrocarbons Promotes Lipid Accumulation and Metabolic Inflammation.

2-naphthol is a low-molecular-weight (LMW) polycyclic aromatic hydrocarbon (PAH) and air pollutant associated with childhood obesity. There has been a recent emergence of studies on the consequences of PAHs on human health. Current epidemiological reports suggest LMW-PAHs may contribute to obesity incidences in children, yet most studies focus on high-molecular-weight PAHs. This study explores 2-naphthol's impact on obesity and obesity-associated metabolic disorders. To investigate 2-naphthol's effect on lipid metabolism and inflammation, we employed 3T3-L1 and BAT1 cell lines to model white and brown adipocytes, respectively, alongside a murine macrophage cell line (RAW264.7). We found that 2-naphthol increased the expression of key adipogenic and lipogenic genes while decreasing lipolytic gene expression in chronically treated 3T3-L1 and BAT1 adipocytes. In addition, chronic 2-naphthol treatment also suppressed adrenergic-stimulated thermogenic gene expression in BAT1 brown adipocytes. In consistence, an increase in lipid accumulation was demonstrated in BODIPY and Oil Red O-stained adipocytes. Additionally, 3T3-L1 adipocytes and RAW264.7 macrophages chronically exposed to 2-naphthol showed upregulated mRNA expression of major inflammatory cytokines (e.g., tumor necrosis factor α (Tnfα), interleukin-1β (Il-1β), and Il-6). In summary, chronic exposure to 2-naphthol stimulates lipid accumulation in adipocytes and inflammation in adipocytes and macrophages. These findings support previous research that demonstrates 2-naphthol has obesogenic potential.

Cichorioside a biocoumarin modulates lipid and glucose storage on 3T3-L1 cell lines: In vitro and in silico approach

Cichorioside a biocoumarin modulates lipid and glucose storage on 3T3-L1 cell lines: In vitro and in silico approach

Investigating the Anti-Aging Effects of Caviar Oil on Human Skin.

As the largest organ of the human body, the skin serves as a critical barrier against environmental damage. However, many factors, such as genetics, sun exposure, and lifestyle choices can lead to skin damage creating wrinkles, sagging, and loss of elasticity. The use of skincare products containing natural ingredients has become increasingly popular as a way to combat the signs of aging. Caviar oil is one such ingredient that has gained attention due to its rich composition of fatty acids, vitamins, and minerals. The objective of this study was to investigate the potential anti-aging effects of caviar oil and to develop a product, Cavi Balm, which could potentially reduce wrinkles and skin sagging. An in vitro model using the 3T3-L1 cell line was employed to assess the effect of caviar oil on adipocyte differentiation. An ex vivo study using human skin tissue was conducted to investigate the impact of caviar oil on collagen and elastin formation and the expression of matrix metalloproteinase-1,2,9 (MMP-1, MMP-2, MMP-9). Furthermore, 102 participants were enrolled in five clinical studies to evaluate the anti-aging efficacy of our product, "Cavi Balm", in facial and neck wrinkles, facial and eye area lifting, and various skin parameters, such as skin moisture, skin elasticity, skin density, skin tightening relief, skin clarity, and skin turnover. In vitro, caviar oil enhanced adipocyte differentiation, and increased lipid accumulation inside the cells. The ex vivo analysis revealed that caviar oil reduced the expression levels of MMP-1, MMP-2, and MMP-9, and increased the formation of elastin and collagen I, III. Moreover, in the clinical study, Cavi Balm improved skin parameters after one-time use, with more significant effects observed after four weeks of usage. Caviar oil has a substantial impact on mitigating skin aging and holds potential for application in anti-aging products.

Allosteric modulation on specific gene mutated Vitamin D receptor by essential PUFAs and its distinct molecular effects on type 2 diabetes mellitus

A serious health threat affecting the T2DM group is evident more cases T2DM are diagnosed. In this research, we choose to research into all of this possible mechanism of 3T3-L1 Cell lines and Molecular Docking studies Schrodinger software identified Vitamin D, Omega-3, and 6 PUFAs (EPA DHA & AA) Compounds of hydrophilic and hydrophobic pocket throughout molecular modeling besides T2DM. A group of three analog VDRs is being developed for discovery treatment with T2DM. Its use as it was agreed to run a molecular cell culture and docking study. Recognize the binding method involving the compound in T2DM through ADME prediction. The molecular dynamics simulation was enhanced by confirmation of the strength of the possible composite binding. Based on the computational results, the Omega-3 and 6 PUFAs compound encourages energy interaction. The composite contains an in vitro anti-diabetic activity; the compounds have clearly shown that they are active on T2DM. Our studies provide vital information on the findings of the bimolecular T2DM inhibitors.

Mice harboring a R133L heterozygous mutation in LMNA exhibited ectopic lipid accumulation, aging, and mitochondrial dysfunction in adipose tissue.

The LMNA gene encodes for the nuclear envelope proteins lamin A and C (lamin A/C). A novel R133L heterozygous mutation in the LMNA gene causes atypical progeria syndrome (APS). However, the underlying mechanism remains unclear. Here, we used transgenic mice (LmnaR133L/+ mice) that expressed a heterozygous LMNA R133L mutation and 3T3-L1 cell lines with stable overexpression of LMNA R133L (by lentiviral transduction) as in vivo and in vitro models to investigate the mechanisms of LMNA R133L mutations that mediate the APS phenotype. We found that a heterozygous R133L mutation in LMNA induced most of the metabolic disturbances seen in patients with this mutation, including ectopic lipid accumulation, limited subcutaneous adipose tissue (SAT) expansion, and insulin resistance. Mitochondrial dysfunction and senescence promote ectopic lipid accumulation and insulin resistance. In addition, the FLAG-mediated pull-down capture followed by mass spectrometry assay showed that p160 Myb-binding protein (P160 MBP; Mybbp1 ), the critical transcriptional repressor of PGC-1α, was bound to lamin A/C. Increased Mybbp1 levels in tissues and greater Mybbp1 -lamin A/C binding in nuclear inhibit PGC-1α activity and promotes mitochondrial dysfunction. Our findings confirm that the novel R133L heterozygous mutation in the LMNA gene caused APS are associated with marked mitochondrial respiratory chain impairment, which were induced by decreased PGC-1α levels correlating with increased Mybbp1a levels in nuclear, and a senescence phenotype of the subcutaneous fat.

Rebamipide treatment ameliorates obesity phenotype by regulation of immune cells and adipocytes.

Obesity is a medical term used to describe an over-accumulation of adipose tissue. It causes abnormal physiological and pathological processes in the body. Obesity is associated with systemic inflammation and abnormalities in immune cell function. Rebamipide, an amino acid derivative of 2-(1H)-quinolinone, has been used as a therapeutic for the protection from mucosal damage. Our previous studies have demonstrated that rebamipide treatment regulates lipid metabolism and inflammation, leading to prevention of weight gain in high-fat diet mice. In this study, mice were put on a high calorie diet for 11 weeks while receiving injections of rebamipide. Rebamipide treatment reduced the body weight, liver weight and blood glucose levels compared to control mice and reduced both glucose and insulin resistance. Fat accumulation has been shown to cause pro-inflammatory activity in mice. Treatment with rebamipide decreased the prevalence of inflammatory cells such as Th2, Th17 and M1 macrophages and increased anti-inflammatory Treg and M2 macrophages in epididymal fat tissue. Additionally, rebamipide addition inhibited adipocyte differentiation in 3T3-L1 cell lines. Taken together, our study demonstrates that rebamipide treatment is a novel and effective method to prevent diet-induced obesity.

Ulvan as a Reducing Agent for the Green Synthesis of Silver Nanoparticles: A Novel Mouthwash

The antibacterial activity of an Ulvan-based silver nanoparticle (AgNP) system was evaluated in the current study. The green synthesis of biogenic silver nanoparticles was conducted using Ulvan, a sulphated polysaccharide extracted from Ulva lactuca. A novel mouthwash containing AgNPs was prepared, and tested for its efficacy and safety. AgNPs were confirmed with spectrophotometric analysis (UV–A visible spectrophotometer), and the characterisation was established with Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and transmission electron microscopy (TEM). The AgNPs were spherical, and their average size was 8–33 nm, as shown via TEM. The antioxidant assay was conducted via DDPH assay, wherein the AgNPs, at a concentration of 50 μL/mL, showed 93.15% inhibition. Furthermore, anticancer activity was tested by evaluating the cell viability utilising the method of an MTT assay on the 3T3-L1 cell lines. AgNPs, at 30 µL/mL, showed maximal cell viability, denoting no cytotoxic effect. The silver-nanoparticle-based mouthrinse, at a concentration of 100 µL/mL, demonstrated antimicrobial activity against Streptococcus mutans, Staphylococcus aureus, Lactobacillus, and Candida albicans. This study shows that mouthwash prepared from the Ulvan-silver nanoparticle system could be a nontoxic and effective oral antimicrobial agent.

Suppressive Effects of Flavonoids on Macrophage-Associated Adipocyte Inflammation in a Differentiated Murine Preadipocyte 3T3-L1 Cells Co-Cultured with a Murine Macrophage RAW264.7 Cells.

The suppressive effects of flavonoids on macrophage-associated adipocyte inflammation in a differentiated murine preadipocyte cell line (3T3-L1) co-cultured with a murine macrophage cell line (RAW264.7) were evaluated. Extracellular lipid accumulation was investigated via Oil Red O staining. The expression levels of adipogenesis- and inflammation-associated proteins, including CCAAT/enhancer-binding protein (C/EBP)-α, inducible nitric oxide synthase (iNOS), C/EBPβ, peroxisome proliferator-activated receptor γ (PPARγ), and cyclooxygenase-2 (COX-2), were determined via Western blotting. Proinflammatory cytokines, including monocyte chemoattractant protein 1 (MCP-1) and interleukin-6 (IL-6), were assessed using enzyme-linked immunosorbent assay kits. We found that silybin, formononetin, and diosmetin inhibited lipid accumulation and production of proinflammatory cytokines in the co-cultures of 3T3-L1 and RAW264.7 cells. Moreover, they inhibited the protein expression of PPARγ, C/EBPα, COX-2, C/EBPβ, and iNOS in the co-cultures of 3T3-L1 and RAW264.7 cells. These data support that silybin, formononetin, and diosmetin inhibit macrophage-associated adipocyte inflammation and lipid accumulation.

Orphan receptor GPR50 attenuates inflammation and insulin signaling in 3T3-L1 preadipocytes.

Type 2 diabetes (T2DM) is characterized by insulin secretion deficiencies and systemic insulin resistance (IR) in adipose tissue, skeletal muscle, and the liver. Although the mechanism of T2DM is not yet fully known, inflammation and insulin resistance play a central role in the pathogenesis of T2DM. G protein-coupled receptors (GPCRs) are involved in endocrine and metabolic processes as well as many other physiological processes. GPR50 (G protein-coupled receptor 50) is an orphan GPCR that shares the highest sequence homology with melatonin receptors. The aim of this study was to investigate the effect of GPR50 on inflammation and insulin resistance in 3T3-L1 preadipocytes. GPR50 expression was observed to be significantly increased in the adipose tissue of obese T2DM mice, while GPR50 deficiency increased inflammation in 3T3-L1 cells and induced the phosphorylation of AKT and insulin receptor substrate (IRS) 1. Furthermore, GPR50 knockout in the 3T3-L1 cell line suppressed PPAR-γ expression. These data suggest that GPR50 can attenuate inflammatory levels and regulate insulin signaling in adipocytes. Furthermore, the effects are mediated through the regulation of the IRS1/AKT signaling pathway and PPAR-γ expression.

Analysis of DYRK1B, PPARG, and CEBPB Expression Patterns in Adipose-Derived Stem Cells from Patients Carrying DYRK1B R102C and Healthy Individuals During Adipogenesis.

Background: Metabolic syndrome (MetS) is a group of signs and symptoms that are associated with a higher risk of type 2 diabetes mellitus and cardiovascular diseases. The major risk factor for developing MetS is abdominal obesity, which is caused by an increase in adipocyte size or quantity. Increased adipocyte quantity is a result of differentiation of stem cells into adipose tissue. Numerous studies have investigated the expression of key transcription factors, including PPARG and CEBPB during adipocyte differentiation in murine cells such as 3T3-L1 cell lines. To better understand the expression changes during the process of fat accumulation in adipose-derived stem cells (ASCs), we compared the expression of DYRK1B, PPARG, and ẟB in ASCs between the patient (harboring DYRK1B R102C) and control (healthy individuals) groups. Methods: Gene expression was evaluated on the eighth day before induction and days 1, 5, and 15 postinduction. The pluripotent capacity of ASCs and the potential for differentiation into adipocytes were confirmed by flow cytometry analysis of surface markers (CD34, CD44, CD105, and CD90), and Oil Red O staining, respectively. The Expression of DYRK1B, PPARG, and CEBPB were assessed by real-time-polymerase chain reaction in patients and normal individuals. The effects of AZ191, a potent small molecule inhibitor on DYRK1B and CEBPB expression in patients' samples were studied. Result: The expression of DYRK1B kinase and transcription factors (CEBPB and PPARG) are higher in ASCs harboring DYRK1B R102C compared with noncarriers on days 5 and 15 during adipocyte differentiation. These proteins may be helpful to elucidate the mechanisms underlying obesity and obesity-related disorders like MetS. Furthermore, the new compound AZ191 exhibited inhibitory activity toward DYRK1B and CEBPB. We suggest that AZ191 may be helpful in defining the potential roles of DYRK1B and CEBPB in adipogenesis.

Isolation and cytotoxic activities of undescribed iridoid and xanthone glycosides from Centaurium erythraea Rafn. (Gentianaceae)

Centaurium erythraea Rafn. (Gentianaceae) is used in internal traditional therapy as an anthelmintic, hypotensive, antipyretic, and antidiabetic. It is used externally for the treatment of wounds. Ursolic acid, maslinic acid, secologanin, secologanin dimethyl acetal, centauroside A, erythraeaxanthone I, erythraeaxanthone II, and demethyleustomin were isolated from aerial parts of Centaurium erythraea and were identified using spectroscopic methods, including NMR and mass spectrometry. The cytotoxic potency of undescribed compounds was evaluated by the XTT assay against human breast cancer MCF-7, MDA-MB-453 and mouse fibroblast 3T3-L1 cell lines. Erythraeaxanthone II was found to have the most potent cytotoxic activity.

Identification of Selective PPAR-γ Modulators by Combining Pharmacophore Modeling, Molecular Docking, and Adipogenesis Assay.

The clinically used glitazones (rosiglitazone and pioglitazone) for type 2 diabetes mellitus therapy have been linked to serious side effects such as fluid retention, congestive heart failure, weight gain, bone loss, and an increased risk of bladder cancer. The complete activation of PPAR-γ receptors in target tissues is linked to these effects. Many studies have demonstrated that partial PPAR-γ activators (GW0072, PAT5A, GQ16) give equivalent therapeutic benefits to full PPAR-γ agonists without the associated side effects. These breakthroughs cleared the path for the development of partial agonists or selective PPAR-γ modulators (SPPARγMs). This study combined pharmacophore modeling, molecular docking, and an adipogenesis experiment to identify thiazolidine analogs as SPPARMs/partial agonists. A custom library of 220 molecules was created and virtual screened to discover 90 compounds as SPPARγMs/ partial agonists. The chosen eight compounds were synthesized and tested for adipogenesis using 3T3L1 cell lines. These compounds' partial agonistic activity was evaluated in 3T3L1 cell lines by comparing their capacity to stimulate PPAR-γ mediated adipogenesis to that of a full agonist, rosiglitazone. The findings of the adipogenesis experiment demonstrate that all eight compounds examined had a partial potential to stimulate adipogenesis when compared to the full agonist, rosiglitazone. The current investigation identified eight possible PPAR-γ partial agonists or SPPARγMs that may be effective in the treatment of type 2 diabetes mellitus.

Chemical composition and nutraceutical characterization of Balanites roxburghii seed oil

Balanites roxburghii is an underutilized species native to India and known for its medicinal and nutritional properties. Seeds possess a high amount of oil, which could be a new source of edible oil. Here we report the compositional analysis of B. roxburghii seed oil (BRSO) and its variability among the South Indian populations. Oil content ranged from 43.2 % to 61.0 %. Oleic, linoleic, palmitic, and stearic were the major fatty acids, and their content varied greatly among the accessions. Carotenoid content was up to 22.6 mg/kg oil. α-Tocopherol ranged between 1.21 and 10.6 mg/100 g, whereas β and γ-tocopherols ranged between 0.89 and 4.98 mg/100 g. β-Sitosterol was the major phytosterol (52.5–147 mg/100 g), and squalene quantity was significant (up to 65.4 mg/100 g). BRSO has no cytotoxic effects on 3T3-L1 cell lines. Results suggest that the BRSO is a significant source of oil and nutraceuticals.

Antidiabetic Activity of Kamdhenu Ark Extracts of Ethnomedicinal plant Penicum millaceum in 3T3 L1 Cell Lines

Diabetes is a “chronic, metabolic disorder characterized by elevated levels of blood glucose (or blood sugar), which leads over time to serious damage to the heart, blood vessels, eyes, kidneys and nerves” according to WHO. Diabetes mellitus (DM) is becoming more prevalent over the world at an astounding level. The consistent use of allopathic medications creates various secondary health impacts, hence there is an urge for adequate natural remedies for its therapy. The potential health benefits of Ethnomedicinal plant Penicum Millaceum (Proso millet) commonly referred as Nutracereals is unexplored. Kamdhenu (Cow) is regarded as sacred animal. Cow urine also cleanses the human body from the inside out by draining out all pollutants, lowering a variety of health risks such as diabetes, obesity and high blood pressure. Present study explores the antidiabetic properties of Kamdhenu Ark extracts of Ethnomedicinal plant Proso Millet, in vitro through α Amylase Inhibition Assay, α Glucosidase Inhibition assay and Glucose up take Assay in 3t3L1 Cell lines. It also emphases on the cytotoxicity Assay on 3t3L1 Cell lines. The Kamdhenu Ark extracts of Penicum Millaceum exhibited prominent inhibitory activity towards α Amylase and α Glucosidase enzymes with IC50 values of 250±0.5μg/ml and 1520±0.2μg/ml respectively. KPM enchanced glucose Uptake by 40.56% and has negligible cytotoxic effect with reference to toxic control in 3t3L1 cell lines model.