AMP-activated protein kinase complexes containing the β2 regulatory subunit are up-regulated during and contribute to adipogenesis

Katwan, Omar J.; Alghamdi, Fatmah; Almabrouk, Tarek Ali Mohamed; Mancini, Sarah J.; Kennedy, Simon; Oakhill, Jonathan S.; Scott, John W.; Salt, Ian P.

doi:10.1042/bcj20180714

Cited by 24 publications

(12 citation statements)

References 52 publications

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“…3T3-L1 fibroblasts were originally obtained from ATCC and are confirmed to be mycoplasma negative. The cells were cultured and differentiated into adipocytes as described previously (30) and used between passages 7 and 12. Experiments were performed at days 8-12 post differentiation.…”

Section: Culture Stimulation and Lysis Of 3t3-l1 Adipocytesmentioning

confidence: 99%

A-769662 inhibits adipocyte glucose uptake in an AMPK-independent manner

Kopietz

Alshuweishi

Bijland

et al. 2021

Biochemical Journal

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Activation of AMP-activated protein kinase (AMPK) is considered a valid strategy for the treatment of type 2 diabetes. However, despite the importance of adipose tissue for whole-body energy homeostasis, the effect of AMPK activation in adipocytes has only been studied to a limited extent and mainly with the AMP-mimetic 5-aminoimidazole-4-carboxamide-1-b-d-ribofuranoside (AICAR), which has limited specificity. The aim of this study was to evaluate the effect of the allosteric AMPK activators A‑769662 and 991 on glucose uptake in adipocytes. For this purpose, primary rat or human adipocytes, and cultured 3T3-L1 adipocytes, were treated with either of the allosteric activators, or AICAR, and basal and insulin-stimulated glucose uptake was assessed. Additionally, the effect of AMPK activators on insulin-stimulated phosphorylation of Akt and Akt substrate of 160 kDa was assessed. Furthermore, primary adipocytes from ADaM site binding drug-resistant AMPKb1 S108A knock-in mice were employed to investigate specificity of the drugs for the observed effects. Our results show that insulin-stimulated adipocyte glucose uptake was significantly reduced by A‑769662 but not 991, yet neither activator had any clear effects on basal or insulin-stimulated Akt/AS160 signaling. The use of AMPKb1 S108A mutant-expressing adipocytes revealed that the observed inhibition of glucose uptake by A‑769662 is most likely AMPK-independent, a finding which is supported by the rapid inhibitory effect A-769662 exerts on glucose uptake in 3T3-L1 adipocytes. These data suggest that AMPK activation per se does not inhibit glucose uptake in adipocytes and that the effects of AICAR and A-769662 are AMPK-independent.

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Section: Culture Stimulation and Lysis Of 3t3-l1 Adipocytesmentioning

confidence: 99%

A-769662 inhibits adipocyte glucose uptake in an AMPK-independent manner

Kopietz

Alshuweishi

Bijland

et al. 2021

Biochemical Journal

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“…Also, berberine was reported to be antidiabetic effects by AMPK activation and decreased HOMA-IR value and insulin resistance [38]. However, the AMPK activation may produce different effects because the activation of different subunits of AMPK has effective at different ability and potential in different tissues [4,39]. Although all the treatments in the current study could not change HOMA-IR levels, these treatments may prevent gluconeogenesis in the liver and improve the pancreatic regeneration by AMPK activation.…”

Section: Discussionmentioning

confidence: 63%

Potential antidiabetic activity of benzimidazole derivative albendazole and lansoprazole drugs in different doses in experimental type 2 diabetic rats

Dik¹,

Coşkun²,

Bahcivan³

et al. 2021

Turk J Med Sci

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IntroductionType 2 diabetes is defined as a chronic inflammatory metabolic disease that is characterized with the impaired insulin effect and high blood glucose level [1]. The disease has degenerative effects on many tissues and a high incidence worldwide. Although many drugs are used for the treatment of this disease, more effective treatment strategies have yet to be found [2,3].Adenosine 5′-monophosphate activated protein kinase (AMPK) is considered to be an important potential therapeutic target for the treatment of type 2 diabetes (T2D). AMPK is defined as serine/threonine protein kinase complex. The kinase consists catalytic α (α1, α2), regulatory β (β1, β2), and regulatory γ (γ1, γ2, γ3) subunits and express in each tissue [3]. Although the isoforms have been reported to have different effects and tissue-specific, the effects of isoforms have not been fully cleared. However, α1 isoform predominates in the liver and adipose tissue; whereas, α2 predominates in the brain, heart, and skeletal muscles [4]. The AMPK acts as a sensor that determines the AMP/ATP or ADP/ATP ratio. The AMPK is activated by phosphorylation of Thr-172 residue in the α subunit and increasing AMP/ATP ratio in the cell [3][4][5][6]. The activated AMPK provides the phosphorylation of key metabolic proteins that inhibits anabolic activities and increase catabolic activities [3]. The AMPK activation suppresses genes mediating gluconeogenesis and lipogenesis in the liver [7], and the activation of AMPK has been reported to increase insulin secretion from pancreatic β cells [3,8]. On the contrary, the excessive increased insulin, leptin and diacylglycerol levels and hyperglycemia, hyperlipidemia inhibit the AMPK activation [6]. Therefore, the AMPK activation increases insulin sensitivity, stimulates glycogen synthesis while inhibits glycolysis. Thus, it is an important Aim: The aim of this study is to determine the effects of different concentrations of albendazole and lansoprazole, which were benzimidazole derivatives, on endocrinologic and biochemical parameters in experimental type 2 diabetic (T2D) rats. Materials and methods:In this study, 46 male Wistar Albino rats were used. Animals were divided as healthy control (0.1 mL/rat/day saline, s.c, n = 6), diabetes control (0.1 mL/rat/day saline, s.c, n = 8), diabetes+low-dose albendazole (5 mg/kg, oral, n = 8), diabetes+highdose albendazole (10 mg/kg, oral n = 8), diabetes+low-dose lansoprazole (15 mg/kg, subcutaneous, n = 8), and diabetes+high-dose lansoprazole (30 mg/kg, subcutaneous, n = 8). All groups were treated for 8 weeks. The blood samples were analyzed by autoanalyzer and ELISA kits for biochemical and endocrinological parameters, respectively.Results: Glucose, HbA1c, triglyceride, low density cholesterol (LDL), leptin, and Homeostatic Model Assessment for insulin resistance (HOMA-IR) levels increased and insulin and HOMA-β levels decreased in the diabetic rats compared to the healthy control group. The glucose, HbA1c, and triglyceride levels were partially decreased; however, insu...

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“…3T3-L1 fibroblasts were originally obtained from ATCC (#CL-173). The cells were cultured and differentiated into adipocytes as described previously [ 47 ] and used between passages 7 and 12. Experiments were performed at days 8–12 post differentiation.…”

Section: Methodsmentioning

confidence: 99%

Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965

Ahwazi

Neopane

Markby

et al. 2021

Biochemical Journal

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SBI-0206965, originally identified as an inhibitor of the autophagy initiator kinase ULK1, has recently been reported as a more potent and selective AMPK inhibitor relative to the widely used, but promiscuous inhibitor Compound C/Dorsomorphin. Here, we studied the effects of SBI-0206965 on AMPK signalling and metabolic readouts in multiple cell types, including hepatocytes, skeletal muscle cells and adipocytes. We observed SBI-0206965 dose dependently attenuated AMPK-activator (991)-stimulated ACC phosphorylation and inhibition of lipogenesis in hepatocytes. SBI-0206965 (≥ 25 μM) modestly inhibited AMPK signalling in C2C12 myotubes, but also inhibited insulin signalling, insulin-mediated/AMPK-independent glucose uptake, and AICA-riboside uptake. We performed an extended screen of SBI-0206965 against a panel of 140 human protein kinases in vitro, which showed SBI-0206965 inhibits several kinases, including members of AMPK-related kinases (NUAK1, MARK3/4), equally or more potently than AMPK or ULK1. This screen, together with molecular modelling, revealed that most SBI-0206965-sensitive kinases contain a large gatekeeper residue with a preference for methionine at this position. We observed that mutation of the gatekeeper methionine to a smaller side chain amino acid (threonine) rendered AMPK and ULK1 resistant to SBI-0206965 inhibition. These results demonstrate that although SBI-0206965 has utility for delineating AMPK or ULK1 signalling and cellular functions, the compound potently inhibits several other kinases and critical cellular functions such as glucose and nucleoside uptake. Our study demonstrates a role for the gatekeeper residue as a determinant of the inhibitor sensitivity and inhibitor-resistant mutant forms could be exploited as potential controls to probe specific cellular effects of SBI-0206965.

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AMP-activated protein kinase complexes containing the β2 regulatory subunit are up-regulated during and contribute to adipogenesis

Cited by 24 publications

References 52 publications

A-769662 inhibits adipocyte glucose uptake in an AMPK-independent manner

A-769662 inhibits adipocyte glucose uptake in an AMPK-independent manner

Potential antidiabetic activity of benzimidazole derivative albendazole and lansoprazole drugs in different doses in experimental type 2 diabetic rats

Investigation of the specificity and mechanism of action of the ULK1/AMPK inhibitor SBI-0206965

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