Journal: Biomedicines

Article Title: Sympathetic Activation Promotes Sodium Glucose Co-Transporter-1 Protein Expression in Rodent Skeletal Muscle

doi: 10.3390/biomedicines12071456

Figure Lengend Snippet: SGLT1 protein levels are elevated in differentiated L6 skeletal cells following SGLT2i but not combined SGLT1/2i. Cells were treated with vehicle, EMPA, or SOTAG after 7 days of differentiation; data represented as means ± SEM; n = 4/group; statistical analysis was conducted using one-way ANOVA; * p = 0.049.

Article Snippet: Based on the previously published literature, the SGLT2i Empagliflozin (EMPA; Ark Pharma Scientific Limited, Wuhan, China) was used at 30 μM for in vitro purposes [ , ].

Techniques:

Journal: Heart Rhythm O2

Article Title: Sodium-glucose cotransporter-2 inhibitor use in type 2 diabetes mellitus is associated with a lower rate of atrial arrhythmias in a hospitalized real-world population

doi: 10.1016/j.hroo.2024.12.004

Figure Lengend Snippet: Study consort diagram. IPTW = inverse probability treatment weighting; PS = propensity score; SGLT2i = sodium-glucose cotransporter-2 inhibitor. (Figure created in Biorender.com.)

Article Snippet: Dr Chew has received a grant in aid for a separate study of SGLT2i from Astra Zeneca.

Techniques:

Journal: Heart Rhythm O2

Article Title: Sodium-glucose cotransporter-2 inhibitor use in type 2 diabetes mellitus is associated with a lower rate of atrial arrhythmias in a hospitalized real-world population

doi: 10.1016/j.hroo.2024.12.004

Figure Lengend Snippet: Baseline characteristics before propensity matching

Article Snippet: Dr Chew has received a grant in aid for a separate study of SGLT2i from Astra Zeneca.

Techniques: Filtration, Medications

Journal: Heart Rhythm O2

Article Title: Sodium-glucose cotransporter-2 inhibitor use in type 2 diabetes mellitus is associated with a lower rate of atrial arrhythmias in a hospitalized real-world population

doi: 10.1016/j.hroo.2024.12.004

Figure Lengend Snippet: Baseline characteristics after inverse probability treatment weighting

Article Snippet: Dr Chew has received a grant in aid for a separate study of SGLT2i from Astra Zeneca.

Techniques: Filtration, Medications

Journal: Heart Rhythm O2

Article Title: Sodium-glucose cotransporter-2 inhibitor use in type 2 diabetes mellitus is associated with a lower rate of atrial arrhythmias in a hospitalized real-world population

doi: 10.1016/j.hroo.2024.12.004

Figure Lengend Snippet: HRs of incidence of cardiac arrhythmias associated with SGLT2i therapy

Article Snippet: Dr Chew has received a grant in aid for a separate study of SGLT2i from Astra Zeneca.

Techniques:

Journal: Heart Rhythm O2

Article Title: Sodium-glucose cotransporter-2 inhibitor use in type 2 diabetes mellitus is associated with a lower rate of atrial arrhythmias in a hospitalized real-world population

doi: 10.1016/j.hroo.2024.12.004

Figure Lengend Snippet: Kaplan-Meier curve showing cumulative incidence of atrial arrhythmias (percentage) over 2 years of follow up. Blue line indicates no sodium-glucose cotransporter-2 inhibitor (SGLT2i) use. Red line indicates SGLT2i use. Hazard ratio (HR) associated 0.17, 95% confidence interval (CI) 0.07–0.41, P <.001.

Article Snippet: Dr Chew has received a grant in aid for a separate study of SGLT2i from Astra Zeneca.

Techniques:

Journal: Heart Rhythm O2

Article Title: Sodium-glucose cotransporter-2 inhibitor use in type 2 diabetes mellitus is associated with a lower rate of atrial arrhythmias in a hospitalized real-world population

doi: 10.1016/j.hroo.2024.12.004

Figure Lengend Snippet: Cumulative incidence of atrial arrhythmias (percentage) over 2 years of follow up according to SGLT2i use. Blue line indicates no SGLT2i use. Red line indicates SGLT2i use. Left: Patients with a previous history of atrial fibrillation (HR 0.35, 95% CI 0.13–0.90, P = .030). Right: Patients without a previous history of atrial fibrillation (HR 0.07, 95% CI 0.01–0.51, P = .009). Abbreviations as in Figure 2 .

Article Snippet: Dr Chew has received a grant in aid for a separate study of SGLT2i from Astra Zeneca.

Techniques:

Journal: Cell Reports

Article Title: Proximal Tubule mTORC1 Is a Central Player in the Pathophysiology of Diabetic Nephropathy and Its Correction by SGLT2 Inhibitors

doi: 10.1016/j.celrep.2020.107954

Figure Lengend Snippet: Effects of Short- and Long-Term Treatment with SGLT2i on Glomerular Size, Renal Injury, and Tubule-Interstitial Fibrosis Akita mice aged 7–8 weeks were treated with and without dapagliflozin (10 mg/kg/day) for 5 days or 3 months. (A) Immunofluorescence staining for the tubule injury marker cystatin-C at the age of 2 months. (B and C) Representative periodic acid-Schiff (PAS) staining and quantifications of glomerular and Bowman’s space cross-sectional areas at the ages of 2 months (B) and 5 months (C). (D and E) Immunofluorescence staining for the fibrosis marker collagen III (D) and the proximal tubule injury marker cystatin-C (E). Representative images and quantification of fluorescence intensity are shown. (F) Representative PAS staining of the glomeruli and quantification of PAS-positive percentage of mesangial area. Scale bar, 50 μm. Data represent the mean ± SEM of six to eight mice per group. ∗∗ p < 0.01 relative to the wild-type control group; ## p < 0.01 relative to the untreated Akita mice group.

Article Snippet: For studying S6 phosphorylation, cells were starved in serum-free DMEM for 1 h and then exposed to either 5 mM or 30 mM D-glucose (low and high glucose, respectively) in the presence or absence of SGLT2i (dapagliflozin; 5 μM, Cayman Chemical) for 0.5 h. For immunofluorescence, cells were exposed to either 5 mM or 30 mM D-glucose in the presence or absence of SGLT2i (5 μM) for 48 h. Rapamycin and Torin-1 (Cell Signaling) were added to the medium at a concentration of 100 and 250 nM, respectively.

Techniques: Immunofluorescence, Staining, Marker, Fluorescence, Control

Journal: Cell Reports

Article Title: Proximal Tubule mTORC1 Is a Central Player in the Pathophysiology of Diabetic Nephropathy and Its Correction by SGLT2 Inhibitors

doi: 10.1016/j.celrep.2020.107954

Figure Lengend Snippet: Effects of Diabetes and of Treatment with SGLT2i on RPTCs mTORC1 Activity (A) Immunofluorescence staining for pS6 on kidney sections of 2-month-old wild-type and Akita mice. (B) Immunofluorescence staining for YFP and pS6 in lineage-traced Sglt2-Cre;Rosa26-YFP + reporter wild-type and Akita mice. (C) Correlation coefficient of YFP and pS6 co-expression. (D and E) pS6 fluorescence intensity (D) and (E) quantifications of RPTC area. (F and G) Immunofluorescence staining for pS6 on kidney sections of streptozotocin (STZ)-induced diabetic mice and in db/db mice (F). Quantification of pS6 fluorescence intensity is shown in (G). (H and I) Immunofluorescence staining for pS6 on kidney sections of wild-type and Akita mice treated with and without dapagliflozin for 12 weeks (H). Insets shown below are higher magnification of the area surrounded by a square; quantifications are shown in (I). (J) Western blotting for pS6 on whole-kidney extracts of wild-type and Akita mice treated with and without dapagliflozin. Scale bar, 50 μm. Data represent the mean ± SEM of six to eight mice per group. ∗ p < 0.05 and ∗∗ p < 0.01 relative to the wild-type control group; ## p < 0.01 relative to the untreated Akita mice group.

Article Snippet: For studying S6 phosphorylation, cells were starved in serum-free DMEM for 1 h and then exposed to either 5 mM or 30 mM D-glucose (low and high glucose, respectively) in the presence or absence of SGLT2i (dapagliflozin; 5 μM, Cayman Chemical) for 0.5 h. For immunofluorescence, cells were exposed to either 5 mM or 30 mM D-glucose in the presence or absence of SGLT2i (5 μM) for 48 h. Rapamycin and Torin-1 (Cell Signaling) were added to the medium at a concentration of 100 and 250 nM, respectively.

Techniques: Activity Assay, Immunofluorescence, Staining, Expressing, Fluorescence, Western Blot, Control

Journal: Cell Reports

Article Title: Proximal Tubule mTORC1 Is a Central Player in the Pathophysiology of Diabetic Nephropathy and Its Correction by SGLT2 Inhibitors

doi: 10.1016/j.celrep.2020.107954

Figure Lengend Snippet: Effects of High Glucose and SGLT2i on mTORC1 Activity, Glycolysis, and Mitochondrial Respiration Cultured RPTCs (LLC-PK1 and HK2) were incubated at 5 mM glucose (low-glu) or 30 mM glucose (high-glu) with or without dapagliflozin (5 μM) for 48 h. The mTORC1 inhibitors rapamycin (rapa) and Torin-1 were used as controls. (A and B) Immunofluorescence staining for pS6 (A) and quantification of fluorescence intensity (B) in LLC-PK1 cells. (C) Western blotting for pS6 in LLC-PK1 cells. (D) Immunofluorescence staining for pS6 and quantification of fluorescence intensity in HK2 cells. (E–G) Oxygen consumption rate (OCR) (E and F) and (G) extracellular acidification rate (ECAR) in LLC-PK1 cells. Scale bar, 50 μm. Data represent the mean ± SEM of three independent experiments. ∗ p < 0.05 and ∗∗ p < 0.01 relative to cells incubated at low glucose; # p < 0.05 and ## p < 0.01 relative to the high-glucose group.

Article Snippet: For studying S6 phosphorylation, cells were starved in serum-free DMEM for 1 h and then exposed to either 5 mM or 30 mM D-glucose (low and high glucose, respectively) in the presence or absence of SGLT2i (dapagliflozin; 5 μM, Cayman Chemical) for 0.5 h. For immunofluorescence, cells were exposed to either 5 mM or 30 mM D-glucose in the presence or absence of SGLT2i (5 μM) for 48 h. Rapamycin and Torin-1 (Cell Signaling) were added to the medium at a concentration of 100 and 250 nM, respectively.

Techniques: Activity Assay, Cell Culture, Incubation, Immunofluorescence, Staining, Fluorescence, Western Blot

Journal: Cell Reports

Article Title: Proximal Tubule mTORC1 Is a Central Player in the Pathophysiology of Diabetic Nephropathy and Its Correction by SGLT2 Inhibitors

doi: 10.1016/j.celrep.2020.107954

Figure Lengend Snippet: Activation of mTORC1 in Akita RPTCs Abrogates the Renal-Protective Effect of SGLT2i RPTC-Tsc1 -KO Akita mice were treated with or without dapagliflozin (10 mg/kg/day in drinking water) for 12 weeks and compared with control Akita mice. (A and B) pS6 expression by immunofluorescence (A) and by western blotting (B). (C and D) Immunofluorescence for cystatin-C (C) and (D) for collagen III. (E–I) Parameters of kidney injury and function: urine ACR (E), urine KIM-1 levels (F), BUN (G), serum creatinine (H), and creatinine clearance (I). Scale bar, 50 μm. Data represent the mean ± SEM of three or four mice per group. ∗ p < 0.05 and ∗∗ p < 0.01 relative to the control Akita group; ## p < 0.01 relative to the Akita - Tsc1 fl/fl group.

Article Snippet: For studying S6 phosphorylation, cells were starved in serum-free DMEM for 1 h and then exposed to either 5 mM or 30 mM D-glucose (low and high glucose, respectively) in the presence or absence of SGLT2i (dapagliflozin; 5 μM, Cayman Chemical) for 0.5 h. For immunofluorescence, cells were exposed to either 5 mM or 30 mM D-glucose in the presence or absence of SGLT2i (5 μM) for 48 h. Rapamycin and Torin-1 (Cell Signaling) were added to the medium at a concentration of 100 and 250 nM, respectively.

Techniques: Activation Assay, Control, Expressing, Immunofluorescence, Western Blot

Journal: Cell Reports

Article Title: Proximal Tubule mTORC1 Is a Central Player in the Pathophysiology of Diabetic Nephropathy and Its Correction by SGLT2 Inhibitors

doi: 10.1016/j.celrep.2020.107954

Figure Lengend Snippet: mTORC1 Regulation of Fibrogenesis, Amino Acid and Glucose Transport, Oxidative Stress, and Pro-inflammatory Genes (A) Gene expression in kidney cortex extracts of 5-month-old RPTC-specific Tsc1-KO ( Sglt2Cre;Tsc1 fl/fl ) mice compared with controls. (B) 4E-BP1 phosphorylation. (C and D) 4E-BP1 phosphorylation (C) and gene expression (D) in kidney cortex extracts of 5-month-old Akita mice and RPTC-specific Raptor-KO ( Sglt2Cre;Raptor fl/+ ) Akita mice. (E and F) Expression of collagen, amino acid transporters (E), and BCAA-degrading enzymes (F) in Akita mice treated with and without dapagliflozin for 3 months compared with wild-type (WT) controls. (G) LLC-PK1 cells treated with 20 mM BCH or left untreated for 3 and 24 h. mTORC1 activity was analyzed using pS6 immunostaining. Quantification of fluorescence intensity is shown. (H) Wild-type and Akita mice were intraperitoneally (i.p.) injected with BCH (2 μM/g body weight per day) for 5 days. mTORC1 activity was analyzed using immunostaining for pS6 and quantification of pS6 fluorescence intensity in RPTCs. (I) A mechanistic model of the effects of diabetes and of treatment with SGLT2i on the development and progression of DKD (see text for details). Scale bar, 50 μm. Data represent the mean ± SEM of three to five mice per group. ∗ p < 0.05 and ∗∗ p < 0.01 relative to the control wild-type or Akita groups; # p < 0.05 and ## p < 0.01 relative to the Akita control group; ππ p < 0.01 relative to the BCH untreated control group.

Article Snippet: For studying S6 phosphorylation, cells were starved in serum-free DMEM for 1 h and then exposed to either 5 mM or 30 mM D-glucose (low and high glucose, respectively) in the presence or absence of SGLT2i (dapagliflozin; 5 μM, Cayman Chemical) for 0.5 h. For immunofluorescence, cells were exposed to either 5 mM or 30 mM D-glucose in the presence or absence of SGLT2i (5 μM) for 48 h. Rapamycin and Torin-1 (Cell Signaling) were added to the medium at a concentration of 100 and 250 nM, respectively.

Techniques: Gene Expression, Phospho-proteomics, Expressing, Activity Assay, Immunostaining, Fluorescence, Injection, Control