A xanthene derivative, DS20060511, attenuates glucose intolerance by inducing skeletal muscle-specific GLUT4 translocation in mice - PubMed (original) (raw)
A xanthene derivative, DS20060511, attenuates glucose intolerance by inducing skeletal muscle-specific GLUT4 translocation in mice
Shinji Furuzono et al. Commun Biol. 2021.
Abstract
Reduced glucose uptake into the skeletal muscle is an important pathophysiological abnormality in type 2 diabetes, and is caused by impaired translocation of glucose transporter 4 (GLUT4) to the skeletal muscle cell surface. Here, we show a xanthene derivative, DS20060511, induces GLUT4 translocation to the skeletal muscle cell surface, thereby stimulating glucose uptake into the tissue. DS20060511 induced GLUT4 translocation and stimulated glucose uptake into differentiated L6-myotubes and into the skeletal muscles in mice. These effects were completely abolished in GLUT4 knockout mice. Induction of GLUT4 translocation by DS20060511 was independent of the insulin signaling pathways including IRS1-Akt-AS160 phosphorylation and IRS1-Rac1-actin polymerization, eNOS pathway, and AMPK pathway. Acute and chronic DS20060511 treatment attenuated the glucose intolerance in obese diabetic mice. Taken together, DS20060511 acts as a skeletal muscle-specific GLUT4 translocation enhancer to facilitate glucose uptake. Further studies of DS20060511 may pave the way for the development of novel antidiabetic medicines.
© 2021. The Author(s).
Conflict of interest statement
The authors declare the following competing interests: S.F., J.T., M.K., A.N., and H.K. are employees of Daiichi Sankyo Co., Ltd. (Tokyo, Japan). The remaining authors declare no competing interests.
Figures
Fig. 1. A xanthene derivative, DS20060511, induced skeletal muscle-specific GLUT4 translocation.
a Chemical structure of DS20060511. b, c Concentration-dependent induction of GLUT4 translocation by DS20060511 and insulin in L6-GLUT4myc myotubes (b) and 3T3-L1-GLUT4myc adipocytes (c). d, e 2-DG uptake evaluated in L6-GLUT4myc myotubes (d) and 3T3-L1-GLUT4myc adipocytes (e). Values shown are means ± SEM, n = 3. **P < 0.01, ***P < 0.001 vs. control by one-way ANOVA followed by Dunnett’s test.
Fig. 2. Treatment with DS20060511 decreases the blood glucose levels via inducing GLUT4 translocation and increasing skeletal muscle glucose uptake.
a, b Blood glucose levels after treatment with DS20060511 (30 mg kg−1) in C57BL/6 mice that had received continued access to food (a) and mice that had been denied access to food overnight (b) (n = 8). Values shown are means ± SEM. **P < 0.01 vs. 0 min by one-way ANOVA followed by Dunnett’s test. c Blood glucose and plasma insulin levels during oral GTT in the C57BL/6 mice (n = 5–6). The mice received oral administration of vehicle or DS20060511 at the indicated dose, 15 min prior to glucose administration (1.5 g kg−1). Values shown are means ± SEM. *P < 0.05, **P < 0.01 vs. vehicle by one-way ANOVA followed by Williams’ test. d [3H]-2-DG uptake in the soleus muscle, gastrocnemius muscle (Gastro.), heart, and white adipose tissue (WAT) at 60 min during the intraperitoneal GTT in the C57BL/6 mice (n = 3). The mice received oral administration of vehicle or DS20060511 (30 mg kg−1), 15 min prior to glucose administration (1 g kg−1 glucose containing [3H]-2-DG). Values shown are means ± SEM. *P < 0.05, **P < 0.01 vs. vehicle by the _t_-test. e Protein levels of GLUT4 and Na,K-ATPaseα in the plasma membrane fraction of the triceps surae muscle excised from the C57BL/6 mice (n = 2) treated with DS20060511 (10 mg kg−1), insulin (5 U kg−1), or saline as vehicle, via the inferior vena cava 10 min after the treatment. Uncropped blots for e can be found in Supplementary Fig. 6.
Fig. 3. The glucose-lowering effect of DS20060511 is totally dependent on GLUT4.
a Blood glucose and plasma insulin levels during oral GTT in wild-type (WT, n = 5) and GLUT4 knockout (KO, n = 6) mice. Mice received oral administration of vehicle or DS20060511 (30 mg kg−1), 15 min prior to the glucose administration (1.5 g kg−1). Values shown are means ± SEM. *P < 0.05, **P < 0.01 vs. vehicle by the _t_-test. b DS20060511-stimulated [3H]-2-DG uptake in the isolated soleus and EDL muscles excised from WT (n = 6) and KO (n = 6) mice. Values shown are means ± SEM. **P < 0.01 vs. vehicle by the _t_-test.
Fig. 4. The mechanism underlying DS20060511-induced GLUT4 translocation is distinct from that of insulin-induced GLUT4 translocation.
a, b Phosphorylation of IRβ, IRS1, Akt (Ser473), and AS160 of the triceps surae muscle excised from C57BL/6 mice (n = 2) treated with DS20060511 (10 mg kg−1), insulin (5 U kg−1), or saline as vehicle, via inferior vena cava 10 min after the treatment. c Fluorescence immunostaining of cell surface GLUT4 and intracellular actin fibers in L6-GLUT4myc myotubes treated with 30 μM of DS20060511 or 100 nM of insulin. Arrowheads indicate the characteristic ruffled structure of the polymerized actin and actin-associated surface GLUT4. d GLUT4 translocation activity of 30 μM DS20060511 or 100 nM insulin in the presence of the actin polymerization inhibitor, Latrunculin B, at the indicated concentrations. Values shown are means ± SEM, n = 3. **P < 0.01 vs. 0 nM Latrunculin B by one-way ANOVA followed by Dunnett’s test. e Concentration-dependent insulin-stimulated GLUT4 translocation in L6-GLUT4myc myotubes with or without 30 μM DS20060511 (n = 3). f Concentration-dependent DS20060511-stimulated 2-DG uptake with 100 nM insulin in isolated muscles from C57BL/6 mice (n = 3). Values shown are means ± SEM. ***P < 0.001 by one-way ANOVA followed by Tukey’s test. g Blood glucose levels during ITT in STZ-treated C57BL/6 mice (n = 6–7). Vehicle or indicated dose of DS20060511 was given orally at the same time as 0.1 U kg−1 insulin injection intraperitoneally. Values shown are means ± SEM. *P < 0.05 vs. vehicle by one-way ANOVA followed by Dunnett’s test. c Scale bar in all panels, 5 μm. Uncropped blots for a and b can be found in Supplementary Fig. 6.
Fig. 5. DS20060511 increases glucose oxidation during exercise.
a–c Respiratory exchange ratio (RER), estimated glucose oxidation, and fat oxidation during stepwise treadmill running in C57BL/6 mice (n = 7). Vehicle or DS20060511 (30 mg kg−1) was given orally 15 min before starting running. Treadmill started from the velocity of 10 m min−1 and increased by 2 m min−1 each 3 min. Values shown are means ± SEM. *P < 0.05 vs. vehicle by the _t_-test.
Fig. 6. DS20060511 increases glucose uptake without affecting AMPK phosphorylation.
a Concentration-dependent DS20060511-stimulated 2-DG uptake with muscle contraction (5 Hz electrical stimulation) in isolated muscles from C57BL/6 mice (n = 3). ***P < 0.001 by one-way ANOVA followed by Tukey’s test. b Muscle contraction (5 Hz electrical stimulation) induced AMPK (Thr172) phosphorylation with or without 10 μM DS20060511 in isolated muscles from C57BL/6 mice. c Phosphorylation levels of AMPKα of the Triceps surae muscles excised from C57BL/6 mice (n = 2) treated with DS20060511 (10 mg kg−1) or saline as vehicle via inferior vena cava 10 min after the treatment. Uncropped blots for b and c can be found in Supplementary Fig. 7.
Fig. 7. DS20060511 treatment decreases blood glucose levels in both WT and eNOSKO mice during oral GTT and ITT.
a, b Blood glucose levels during oral GTT in wild-type (WT, n = 5) and eNOS-knockout (KO, n = 5–6) mice. Mice received vehicle or DS20060511 (10 mg kg−1) orally 15 min before the glucose administration (3.0 g kg−1). c, d Blood glucose levels during ITT in WT (n = 4) and KO (n = 5) mice. Vehicle or DS20060511 (30 mg kg−1) was given orally at the same time as 0.5 U kg−1 insulin injection intraperitoneally. Values shown are means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. vehicle by the _t_-test.
Fig. 8. Acute and chronic DS20060511 treatment improves glucose intolerance in HFD-fed and db/db mice.
a Blood glucose and plasma insulin levels during an oral GTT in normal-chow (NC)- and high-fat diet (HFD)-fed mice (n = 5). Vehicle or DS20060511 (30 mg kg−1) was given orally 15 min before the oral glucose administration (1.5 g kg−1). Values shown are means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. the HFD vehicle by the _t_-test. b Effects of 10 μM DS20060511 and 100 nM insulin on 2-DG uptake in muscles isolated from NC-fed (n = 6) and HFD-fed (n = 5) mice. Values shown are means ± SEM. **P < 0.01 by one-way ANOVA followed by Tukey’s test. c, d Changes in the blood glucose levels and AUC on day 1 and day 28 during refeeding (n = 6) in db/db mice treated chronically with DS20060511 (10 mg kg−1 day−1). Values shown are means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. vehicle by the _t_-test. e Change of the HbA1c levels in the db/db (n = 6) mice at 4 weeks. Values shown are means ± SEM. *P < 0.05 vs. vehicle by the _t_-test.
References
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Research Materials