scholarly journals α-Poly-l-lysine functions as an adipogenic inducer in 3T3-L1 preadipocytes

Amino Acids ◽  
2021 ◽  
Author(s):  
Kyeong Won Lee ◽  
Young Jun An ◽  
Janet Lee ◽  
Jung-Hyun Lee ◽  
Hyung-Soon Yim
Keyword(s):  
Molecular Weight ◽  
Insulin Signaling ◽  
Adipocyte Differentiation ◽  
Cell Attachment ◽  
Underlying Mechanism ◽  
High Concentration ◽  
Expansion Phase ◽  
Regulatory Pathways ◽  
Mitotic Clonal Expansion ◽  
Positive Effect

Abstractα-Poly-l-lysine (PLL) has been used for various purposes such as cell attachment, immunization, and molecular delivery, and is known to be cytotoxic to several cell lines. Here, we studied the effect of PLL on the adipogenesis of 3T3-L1 cells and investigated the underlying mechanism. Differentiation media containing PLL with a molecular weight (MW) greater than 4 kDa enhanced lipid droplet formation and increased adipogenic marker levels, indicating an increase in adipocyte differentiation. PLL with a molecular weight between 30 and 70 kDa was more effective than PLL of other sizes in 3T3-L1 cell differentiation. Moreover, PLL induced 3T3-L1 adipogenesis in insulin-free adipocyte differentiation medium. Incubation with insulin and PLL exhibited greater adipogenesis than insulin treatment only even at a high concentration. PLL stimulated insulin signaling and augmented the signaling pathway when it was added with insulin. While PLL did not activate the glucocorticoid receptor, which is phosphorylated by dexamethasone (DEX), it showed a positive effect on the cAMP signal pathway when preadipocytes were treated with PLL and 3-isobutyl-1-methylxanthine (IBMX). Consistent with these results, incubation with PLL and DEX without IBMX induced adipocyte differentiation. We also observed that the mitotic clonal expansion phase was the critical stage in adipogenesis for inducing the effects of PLL. These results suggest that PLL functions as an adipogenic inducer in 3T3-L1 preadipocytes and PLL has a direct effect on insulin signaling, one of the main regulatory pathways.

scholarly journals A Synergistic Anti-Diabetic Effect by Ginsenosides Rb1 and Rg3 through Adipogenic and Insulin Signaling Pathways in 3T3-L1 Cells

2021 ◽  
Vol 11 (4) ◽  
pp. 1725
Author(s):  
Hee-Do Hong ◽  
Sun-Il Choi ◽  
Ok-Hwan Lee ◽  
Young-Cheul Kim
Keyword(s):  
Transcription Factors ◽  
Insulin Signaling ◽  
Adipocyte Differentiation ◽  
Red Ginseng ◽  
Inhibitory Effects ◽  
Rt Pcr ◽  
High Concentration ◽  
Expression Of Genes ◽  
The Individual ◽  
Maximal Expression

Although ginsenosides Rb1 and Rg3 have been identified as the significant ginsenosides found in red ginseng that confer anti-diabetic actions, it is unclear whether insulin-sensitizing effects are mediated by the individual compounds or by their combination. To determine the effect of ginsenosides Rb1 and Rg3 on adipocyte differentiation, 3T3-L1 preadipocytes were induced to differentiate the standard hormonal inducers in the absence or presence of ginsenosides Rb1 or Rg3. Additionally, we determined the effects of Rb1, Rg3, or their combination on the expression of genes related to adipocyte differentiation, adipogenic transcription factors, and the insulin signaling pathway in 3T3-L1 cells using semi-quantitative RT-PCR. Rb1 significantly increased the expression of CEBPα, PPARγ, and aP2 mRNAs. However, Rg3 exerted its maximal stimulatory effect on these genes at 1 μM concentration, while a high concentration (50 μM) showed inhibitory effects. Similarly, treatment with Rb1 and Rg3 (1 μM) increased the expression of IRS-1, Akt, PI3K, GLUT4, and adiponectin. Importantly, co-treatment of Rb1 and Rg3 (9:1) induced the maximal expression levels of these mRNAs. Our data indicate that the anti-diabetic activity of red ginseng is, in part, mediated by synergistic actions of Rb1 and Rg3, further supporting the significance of minor Rg3.

scholarly journals The nature of conditioning nutrients for human malignant melanoma cultures

1983 ◽  
Vol 62 (1) ◽  
pp. 249-266
Author(s):  
K.A. Ellem ◽  
G.F. Kay
Keyword(s):  
Molecular Weight ◽  
Cell Density ◽  
Cell Attachment ◽  
Cell Types ◽  
Fresh Medium ◽  
Low Molecular Weight ◽  
High Concentration ◽  
Cell Content ◽  
Human Malignant Melanoma ◽  
Serum Factors

From a human melanoma line (MM96), showing some dependence of its rate of growth and cell attachment on serum concentration, sublines were selected for even greater dependence on serum factors. These sublines were used to identify the production of substances by other melanoma cells in culture that would supplement or replace the requirement for serum. Most of the sublines showed higher colony-forming efficiency in medium conditioned by one of several cell types in the presence of a low concentration of serum (2.5%) compared with fresh medium containing a high concentration of serum (10%). The conditioning factor(s) were found to be moderately heat-stable, nonlipophilic, and to be of low molecular weight (less than or greater than 400). Screening of a variety of non-essential low molecular weight nutrients, which have been reported to potentiate the growth of a variety of cell types in low-density culture, was positive for the MM96 sublines only for pyruvate. In particular, L-alanine, L-serine, putrescine and alpha MSH (melanocyte-stimulating hormone) were ineffective. Despite the problems of comparing conditioned media with fresh medium, a reasonable correlation between the stimulatory effect and the cell content of added 2-oxocarboxylates was apparent. As would be anticipated, MM96 cultures showed a population density-dependent enhancement of growth up to a cell density of 2 to 4 × 10(4) cells cm-2. Further increase in the initial cell density of these cultures led to a decline in growth rate. An important additional observation was that simple dilution of the ingredients of RPMI1640 with phosphate-buffered saline or Hanks' balanced salt solution led to a reversal of growth inhibition accompanying a serum shift-down.

scholarly journals Curcumin Attenuates Adipogenesis by Inducing Preadipocyte Apoptosis and Inhibiting Adipocyte Differentiation

Nutrients ◽  
2019 ◽  
Vol 11 (10) ◽  
pp. 2307 ◽  
Author(s):  
Liang-Yi Wu ◽  
Chien-Wei Chen ◽  
Luen-Kui Chen ◽  
Hsiang-Yun Chou ◽  
Chih-Ling Chang ◽  
...  
Keyword(s):  
Metabolic Syndrome ◽  
Lipid Accumulation ◽  
Adipocyte Differentiation ◽  
Underlying Mechanism ◽  
Tunel Assay ◽  
Caspase Activation ◽  
Increased Risk ◽  
Diphenyltetrazolium Bromide ◽  
Mitotic Clonal Expansion

Patients with metabolic syndrome are at an increased risk of developing type 2 diabetes and cardiovascular diseases. The principal risk factor for development of metabolic syndrome is obesity, defined as a state of pathological hyperplasia or/and hypertrophy of adipose tissue. The number of mature adipocytes is determined by adipocyte differentiation from preadipocytes. The purpose of the present study is to investigate the effects of curcumin on adipogenesis and the underlying mechanism. To examine cell toxicity of curcumin, 3T3-L1 preadipocytes were treated with 0–50 µM curcumin for 24, 48, or 72 h, then cell viability was measured using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The effect of curcumin on the cell cycle was determined by flow cytometry. Curcumin-induced cell apoptosis was determined by the TUNEL assay and curcumin-induced caspase activation was measured by immunoblotting. The effect of curcumin on adipocyte differentiation was determined by measuring mitotic clonal expansion (MCE), expression of adipogenic transcription factors, and lipid accumulation. Results showed the viability of preadipocytes was significantly decreased by treatment with 30 µM curcumin, a concentration that caused apoptosis in preadipocytes, as assessed by the TUNEL assay, and caused activation of caspases 8, 9, and 3. A non-cytotoxic dose of curcumin (15 µM) inhibited MCE, downregulated the expression of PPARγ and C/EBPα, prevented differentiation medium-induced β-catenin downregulation, and decreased the lipid accumulation in 3T3-L1 adipocytes. In conclusion, our data show that curcumin can induce preadipocyte apoptosis and inhibit adipocyte differentiation, leading to suppression of adipogenesis.

Metathesis Polymerization of Phenylacetylene by Tungsten Aryloxo Complexes

1995 ◽  
Vol 60 (3) ◽  
pp. 489-497 ◽  
Author(s):  
Hynek Balcar ◽  
Jan Sedláček ◽  
Marta Pacovská ◽  
Vratislav Blechta
Keyword(s):  
Molecular Weight ◽  
Catalytic Activity ◽  
Induction Period ◽  
Average Molecular Weight ◽  
Molar Fraction ◽  
Molar Ratio ◽  
Weight Average Molecular Weight ◽  
Polymer Yield ◽  
Positive Effect ◽  
Ligand Addition

Catalytic activity of the tungsten aryloxo complexes WCl5(OAr) and WOCl3(OAr), where Ar = 4-t-C4H9C6H4, 2,6-(t-C4H9)2C6H3, 2,6-Cl2C6H3, 2,4,6-Cl3C6H2, and 2,4,6-Br3C6H2 in polymerization of phenylacetylene (20 °C, monomer to catalyst molar ratio = 1 000) was studied. The activity of WCl5(OAr) as unicomponent catalysts increases with increasing electron withdrawing character of the -OAr ligand. Addition of two equivalents of organotin cocatalysts (Me4Sn, Bu4Sn, Ph4Sn, Bu3SnH) to WCl5(O-C6H2Cl3-2,4 ,6) has only slight positive effect (slightly higher polymer yield and/or molecular weight of poly(phenylacetylene)s was achieved). However, in the case of WOCl3(O-C6H3Cl2-2, 6) catalyst, it enhances the activity considerably by eliminating the induction period. Poly(phenylacetylene)s prepared with the catalysts studied have weight-average molecular weight ranging from 100 000 to 200 000. They are trans-prevailing and have relatively low molar fraction of monomer units comprised in cyclohexadiene sequences (about 6%).

Metabolomics strategy reveals the osteogenic mechanism of yak (Bos grunniens) bone collagen peptides on ovariectomy-induced osteoporosis in rats

2020 ◽  
Vol 11 (2) ◽  
pp. 1498-1512 ◽  
Author(s):  
Mengliang Ye ◽  
Chunhui Zhang ◽  
Wei Jia ◽  
Qingshan Shen ◽  
Xiaojie Qin ◽  
...  
Keyword(s):  
Underlying Mechanism ◽  
Bone Collagen ◽  
Targeted Metabolomics ◽  
First Report ◽  
Bos Grunniens ◽  
Collagen Peptides ◽  
Positive Effect

This is the first report on the positive effect and underlying mechanism of yak bone collagen peptides in the treatment of osteoporotic rats based on non-targeted metabolomics.

Compatibility Study of Condensate and Heavy Oil for Storage in an Iranian Reservoir

2021 ◽  
pp. 1-13
Author(s):  
K. Zobeidi ◽  
M. Ganjeh-Ghazvini ◽  
V. Hematfar
Keyword(s):  
Heavy Oil ◽  
Oil And Gas ◽  
Production Performance ◽  
Compatibility Study ◽  
Asphaltene Precipitation ◽  
High Concentration ◽  
Full Capacity ◽  
Potential Risks ◽  
Operational Criteria ◽  
Positive Effect

Summary During the years 2017–2020, when Iran faced restrictions on the sale of oil and gas condensate and due to the need for domestic consumption and gas sales commitments, it was inevitable to produce gas at full capacity. This coercion has led to significant production of gas condensates. Some of these condensates were sold, some were converted into products such as gasoline in domestic refineries, and some of these condensates needed to be stored, but the storage capacity was limited. For the purpose of underground condensate storage, a heavy oil reservoir was selected based on some technical and operational criteria. A feasibility study was conducted to evaluate the potential risks of condensate injection into the reservoir. The results of tests on asphaltene precipitation, as the most important risk, indicated no severe precipitation would occur even if high concentration of condensate mixed with the reservoir heavy oil. The recovery of condensate and the production performance of the reservoir were simulated in three different injection-production scenarios. The results showed a positive effect of condensate injection on production rate of the reservoir. Moreover, satisfactory volume of condensate could be recovered in a reasonable period of time.

Abstract 1376: The Nadph Oxidase Nox4 Controls Mkp1 Expression and Thereby the Development of White Adipose Tissue

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Katrin Schröder ◽  
Judith G Schreiber ◽  
Ralf P Brandes
Keyword(s):  
Adipose Tissue ◽  
Risk Factor ◽  
Cardiovascular Risk ◽  
Nadph Oxidase ◽  
Opposite Effect ◽  
Adipocyte Differentiation ◽  
Underlying Mechanism ◽  
3T3 Fibroblasts ◽  
Proliferation And Differentiation

Obesity is a cardiovascular risk factor. Insulin promotes the formation of adipocytes by promoting fibroblast into adipocyte differentiation. Insulin however also acts as a mitogen for fibroblasts and the balance of insulin-induced proliferation and differentiation is incompletely understood. We hypothesize that the NADPH oxidase Nox4 acts as a central switch between these two processes. In 3T3-fibroblasts Nox4 siRNA reduced the cellular radical formation and attenuated the insulin-stimulated differentiation from fibroblasts into adipocytes. Importantly, insulin-induced proliferation was enhanced by Nox4 siRNA. Accordingly, Nox4 overexpression enhanced differentiation and reduced insulin-stimulated proliferation. We generated Nox4 knockout mice to study the in vivo relevance of this observation: The amount of adipose tissue was reduced in the Nox4−/− animals as compared to their wildtype littermates suggesting indeed an attenuated adipocyte differentiation, whereas fibroblasts obtained from Nox4−/− mice presented with enhanced proliferation. To uncover the underlying mechanism, we focused on the Erk1/2 pathway. Erk1/2 signaling is prevented by dephosphorylation through the dualspecific phosphatase MKP1. Luciferase reportergene assays using the full-length MKP1 promotor revealed that Nox4 siRNA reduced, and overexpression of Nox4 as well as treatment of the cells with H 2 O 2 induced MKP1 promotor activity. Indeed, Nox4 siRNA reduced MKP1 protein expression and thus enhanced basal and insulin-induced activation of Erk1/2 in fibroblasts. Nox4 overexpression had the opposite effect. Importantly, MKP1 expression was also reduced in adipose tissue from Nox4−/− mice. Overexpression of MKP1 in fibroblastes increased insulin-induced differentiation and attenuated proliferation, whereas MKP1 siRNA had the opposite effect. We conclude that Nox4, by regulating MKP1, is essentially involved in adipocyte differentiation and development of obesity.

STUDIES ON SMALL MOLECULAR WEIGHT ADHESION PROTEINS (SAPs) FROM CONNECTIVE TISSUES

1987 ◽  
Author(s):  
S Wasi ◽  
P Alles ◽  
D Gauthier ◽  
U Bhargava ◽  
J Farsi ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Polyclonal Antibody ◽  
Type I Collagen ◽  
Cell Attachment ◽  
Binding Capacity ◽  
Guanidine Hydrochloride ◽  
Cell Types ◽  
Type I ◽  
Cell Binding ◽  
Connective Tissues

We have identified a family of low molecular weight proteins with cell attachment properties in a variety of soft and mineralised connective tissues (Wong et al., Biochem. J. 232, 119, 1985). For further characterisation of these proteins we extracted porcine bones with 4 M guanidine hydrochloride and purified the proteins on a series of gel filtration columns The purifed SAPs comprise three bands with Mr -14 000 -17 000. All three proteins bound to heparin-sepahrose in both the presence and absence of 4M urea, and when eluted with 2 M NaCl they retained their cell binding capacity. These proteins promoted the adhesion and spreading of a variety of cell types, including normal fibroblasts, osteoblasts, and epithelial cells, and tumour (osteosarcoma) cells. On Western blotting SAPs did not cross-react with antibodies against fibronectin, laminin or type I collagen; however, they were recognised by a monoclonal antibody to human vitronectin, a polyclonal antibody to bovine vitronectin and polyclonal antibody to human somatomedin B. Dose response experiments indicated that maximum attachment of human gingival fibroblasts occurred in the presence or absence of fetal bovine serum on wells precoated with 2.5 μg/cm2 of SAPs. Attachment of cells to these proteins was partially inhibited by the synthetic pentapeptide Gly-Arg-Gly-Asp-Ser. Utilising the nitrocellulose cell binding assay of Hayman et al (J. Cell. Biol. 95, 20, 1982), the cell attachment to these proteins could be completely inhibited by heparin (100 units/mL) whereas up to 1000 units/mL of heparin had no inhibitory effect on cell attachment to fibronectin and vitronectin. The occurrence of these proteins in a variety of connective tissues and their recognition by different cell types may reflect their general biological role in adhesive mechanisms in both hard and soft connective tissues. Currently, we are investigating the relationship between SAPs and vitronectin, since it is possible that SAPs represent a tissue-processed form of vitronectin or may be novel attachment proteins with regions of homology with vitronectin

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