Inhibition of apoB secretion from HepG2 cells by insulin is amplified by naringenin, independent of the insulin receptor

Wu-Mei-Wan (WMW) is a Chinese herbal formula used to treat type 2 diabetes. In this study, we aimed to explore the effects and mechanisms of WMW on insulin resistance in HepG2 cells. HepG2 cells were pretreated with palmitate (0.25 mM) to impair the insulin signaling pathway. Then, they were treated with different doses of WMW-containing medicated serum and stimulated with 100 nM insulin. Results showed that palmitate could reduce the glucose consumption rate in HepG2 cells and impair insulin signaling related to phosphorylation of insulin receptor (IR) and insulin receptor substrate-1 (IRS-1), thereby regulating the downstream signaling pathways. However, medicated serum of WMW restored impaired insulin signaling, upregulated the expression of phospho-IR (pIR), phosphatidylinositol 3-kinase p85 subunit, phosphoprotein kinase B, and glucose transporter 4, and decreased IRS serine phosphorylation. In addition, it decreased the expression of interleukin-1β and tumor necrosis factor-α, which are the key proinflammatory cytokines involved in insulin resistance; besides, it reduced the expression of NLRP3 inflammasome. These results suggested that WMW could alleviate palmitate-induced insulin resistance in HepG2 cells via inhibition of NLRP3 inflammasome and reduction of proinflammatory cytokine production.

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Selective Attenuation of Metabolic Branch of Insulin Receptor Down-signaling by High Glucose in a Hepatoma Cell Line, HepG2 Cells

Journal of Biological Chemistry ◽

10.1074/jbc.m905410199 ◽

2000 ◽

Vol 275 (27) ◽

pp. 20880-20886 ◽

Cited By ~ 74

Author(s):

Koji Nakajima ◽

Keishi Yamauchi ◽

Satoshi Shigematsu ◽

Sachiko Ikeo ◽

Mitsuhisa Komatsu ◽

...

Keyword(s):

Insulin Receptor ◽

Cell Line ◽

High Glucose ◽

Hepg2 Cells ◽

Hepatoma Cell ◽

Hepatoma Cell Line ◽

Hepatoma Cell Line Hepg2

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911 Iron and insulin resistance: effect of iron manipulation on insulin receptor in HEPG2 cells

Hepatology ◽

10.1016/s0270-9139(03)80950-3 ◽

2003 ◽

Vol 38 ◽

pp. 597-597

Author(s):

P DONGIOVANNI ◽

L VALENTI ◽

L CARMAGNOLA ◽

F DICATERINO ◽

A FRACANZANI ◽

...

Keyword(s):

Insulin Resistance ◽

Insulin Receptor ◽

Hepg2 Cells

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Insulin receptor is phosphorylated in response to treatment of HepG2 cells with insulin-like growth factor I

Biochemical Journal ◽

10.1042/bj2700027 ◽

1990 ◽

Vol 270 (1) ◽

pp. 27-32 ◽

Cited By ~ 7

Author(s):

V Duronio

Keyword(s):

Growth Factor ◽

Insulin Receptor ◽

Hepg2 Cells ◽

Insulin Binding ◽

Response To Treatment ◽

Insulin Like Growth Factor ◽

Factor I ◽

Igf I ◽

Growth Factor I ◽

In Cells

1. Binding of insulin and insulin-like growth factor I (IGF-I) to HepG2 cells was analysed with regard to competition by both insulin and IGF-I. At concentrations of insulin that caused maximal phosphorylation of the insulin receptor, virtually no displacement of IGF-I binding was observed. Similarly, at concentrations of IGF-I that caused maximal phosphorylation of the IGF-I receptor, no displacement of insulin binding was observed. 2. When the phosphorylation of both receptors was examined individually by using specific monoclonal antibodies to immunoprecipitate the receptors, phosphorylation of the insulin receptor was found to increase on both serine and tyrosine residues in cells treated with 100 ng of IGF-I/ml. In contrast, no increased phosphorylation of IGF-I receptor was observed in cells treated with 100 ng of insulin/ml. 3. The increase in phosphorylation of insulin receptor in response to IGF-I correlated with the dose-response of IGF-I-stimulated phosphorylation of the IGF-I receptor. 4. The IGF-I-stimulated phosphorylation of the insulin receptor could be blocked by preincubation with a monoclonal antibody that blocks IGF-I binding to the IGF-I receptor.

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Glucosylceramide synthase inhibitors D-PDMP and D-EtDO-P4 decrease the GM3 ganglioside level, differ in their effects on insulin receptor autophosphorylation but increase Akt1 kinase phosphorylation in human hepatoma HepG2 cells.

Acta Biochimica Polonica ◽

10.18388/abp.2014_930 ◽

2016 ◽

Vol 63 (2) ◽

Cited By ~ 2

Author(s):

Natalia Fedoryszak-Kuśka ◽

Mirosława Panasiewicz ◽

Hanna Domek ◽

Tadeusz Pacuszka

Keyword(s):

Insulin Receptor ◽

Hepg2 Cells ◽

Culture Medium ◽

Control Level ◽

Glucosylceramide Synthase ◽

Control Value ◽

Human Hepatoma Hepg2 Cells ◽

High Level ◽

Kinase Phosphorylation ◽

Hepatoma Hepg2

Gangliosides function as modulators of several cell growth related receptors. It was shown for caveolin-rich adipocytes, that GM3 ganglioside binds to insulin receptor (IR), dissociates its complex with caveolin, and thus lowers IR autophosphorylation following insulin treatment. We extended those studies into human hepatocyte-derived HepG2 cells, characterized by a high level of IR but low of caveolin. To lower the glycosphingolipid content, estimated by GM3 concentration, two glucosylceramide synthase inhibitors d-threo-1-pheny-2-decanoylamino-3-morpholino-1-propanol (d-PDMP) and d-threo-1-(3,4,-ethylenedioxy)phenyl-2-palmitoylamino-3-pyrrolidino-1-propanol (d-EtDO-P4) were used. d-PDMP at 40 µM or d-EtDO-P4 at 1 µM concentrations in culture medium decreased the GM3 content to 22.3% (17.8-26.1%) and 18.1% (13.7-24.4%), respectively, of the control value. The reduction of GM3 obtained with d-PDMP was accompanied by a 185.1% (153.5-423.8%) significant increase in the level of IR autophosphorylation following cell stimulation with 100 nM insulin. The effect of d-EtDO-P4 on IR autophosphorylation was smaller amounting to an increase by 134.8% (111.3-167.8%) of the control level and statistically non-significant. The effects of d-PDMP and d-EtDO-P4 could also be detected at the level of Akt1 kinase. In cells grown in the presence of d-PDMP the level of phosphorylated Akt1 was 286.0% (151.4%-621.1%) of that in the control. In this case the effect of d-EtDO-P4 was similar: 223.0% (181.4-315.4%) significant increase in phosphorylated Akt1. We assume that glycosphingolipid depletion in HepG2 cells may affect not only IR autophosphorylation but also, independently, the phosphorylation of Akt1, by modifying the membrane microenvironment of this kinase.

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Effects of media conditions, insulin, and dexamethasone on insulin-receptor mRNA and promoter activity in HepG2 cells

Diabetes ◽

10.2337/diabetes.40.1.58 ◽

1991 ◽

Vol 40 (1) ◽

pp. 58-65 ◽

Cited By ~ 6

Author(s):

J. R. Levy ◽

G. Krystal ◽

P. Glickman ◽

F. Dastvan

Keyword(s):

Insulin Receptor ◽

Hepg2 Cells ◽

Promoter Activity ◽

Receptor Mrna ◽

Effects Of Media

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Naringenin, a citrus flavonoid, inhibits apoB secretion from HepG2 cells, an effect associated with selective inhibition of ACAT-2 expression

Atherosclerosis ◽

10.1016/s0021-9150(00)81313-1 ◽

2000 ◽

Vol 151 (1) ◽

pp. 289

Author(s):

NicaM. Borradaile ◽

LisaJ. Wilcox ◽

LindaE. de Dreu ◽

MurrayW. Huff

Keyword(s):

Hepg2 Cells ◽

Selective Inhibition ◽

Apob Secretion

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Carpachromene Ameliorates Insulin Resistance in HepG2 Cells via Modulating IR/IRS1/PI3k/Akt/GSK3/FoxO1 Pathway

Molecules ◽

10.3390/molecules26247629 ◽

2021 ◽

Vol 26 (24) ◽

pp. 7629

Author(s):

Rania Alaaeldin ◽

Iman A. M. Abdel-Rahman ◽

Heba Ali Hassan ◽

Nancy Youssef ◽

Ahmed E. Allam ◽

...

Keyword(s):

Insulin Resistance ◽

Enzyme Activity ◽

Insulin Receptor ◽

Glucose Concentration ◽

Hepg2 Cells ◽

Phosphoenolpyruvate Carboxykinase ◽

Glycogen Content ◽

Insulin Signalling ◽

Antidiabetic Activity ◽

Insulin Resistant

Insulin resistance contributes to several disorders including type 2 diabetes and cardiovascular diseases. Carpachromene is a natural active compound that inhibits α-glucosidase enzyme. The aim of the present study is to investigate the potential activity of carpachromene on glucose consumption, metabolism and insulin signalling in a HepG2 cells insulin resistant model. A HepG2 insulin resistant cell model (HepG2/IRM) was established. Cell viability assay of HepG2/IRM cells was performed after carpachromene/metformin treatment. Glucose concentration and glycogen content were determined. Western blot analysis of insulin receptor, IRS1, IRS2, PI3k, Akt, GSK3, FoxO1 proteins after carpachromene treatment was performed. Phosphoenolpyruvate carboxykinase (PEPCK) and hexokinase (HK) enzymes activity was also estimated. Viability of HepG2/IRM cells was over 90% after carpachromene treatment at concentrations 6.3, 10, and 20 µg/mL. Treatment of HepG2/IRM cells with carpachromene decreased glucose concentration in a concentration- and time-dependant manner. In addition, carpachromene increased glycogen content of HepG2/IRM cells. Moreover, carpachromene treatment of HepG2/IRM cells significantly increased the expression of phosphorylated/total ratios of IR, IRS1, PI3K, Akt, GSK3, and FoxO1 proteins. Furthermore, PEPCK enzyme activity was significantly decreased, and HK enzyme activity was significantly increased after carpachromene treatment. The present study examined, for the first time, the potential antidiabetic activity of carpachromene on a biochemical and molecular basis. It increased the expression ratio of insulin receptor and IRS1 which further phosphorylated/activated PI3K/Akt pathway and phosphorylated/inhibited GSK3 and FoxO1 proteins. Our findings revealed that carpachromene showed central molecular regulation of glucose metabolism and insulin signalling via IR/IRS1/ PI3K/Akt/GSK3/FoxO1 pathway.

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