Journal: Cells

Article Title: Dapagliflozin Restores Impaired Autophagy and Suppresses Inflammation in High Glucose-Treated HK-2 Cells

doi: 10.3390/cells10061457

Figure Lengend Snippet: Sequences of the small interfering RNAs used in this study.

Article Snippet: The SGLT2 inhibitor dapagliflozin (DAPA, AstraZeneca, Cambridge, UK) was used at a final concentration of 10–100 μM for 48 h.

Techniques:

Journal: Cells

Article Title: Dapagliflozin Restores Impaired Autophagy and Suppresses Inflammation in High Glucose-Treated HK-2 Cells

doi: 10.3390/cells10061457

Figure Lengend Snippet: Antibodies used for western blotting and immunofluorescence.

Article Snippet: The SGLT2 inhibitor dapagliflozin (DAPA, AstraZeneca, Cambridge, UK) was used at a final concentration of 10–100 μM for 48 h.

Techniques: Western Blot, Immunofluorescence

Journal: Cells

Article Title: Dapagliflozin Restores Impaired Autophagy and Suppresses Inflammation in High Glucose-Treated HK-2 Cells

doi: 10.3390/cells10061457

Figure Lengend Snippet: Dapagliflozin and SGLT2 inhibition suppressed glucose uptake in HK-2 cells. ( A ) HK-2 cells were incubated with Cont or HG (30 mM) medium and treated with or without dapagliflozin (20 µM) for 48 h. SGLT2 was assessed by western blot densitometric evaluation (n = 3) of ( A ). ( B ) Glucose uptake analysis of 2-DG by dapagliflozin-treated HK2 cells (n = 6). ( C ) HK-2 cells were transfected with control siRNA or SGLT2 siRNA (200 nM) for 6 h and then, the medium was replaced with fresh medium with or without HG (30 mM) for 48 h. The SGLT2 level was assessed by western blotting (n = 3). ( D ) Glucose uptake analysis of 2-DG by SGLT2-knockdown HK-2 cells (n = 6). All data represent the means ± standard deviation (SD). * p < 0.05 vs. the indicated group, ** p < 0.01 vs. the indicated group, *** p < 0.001 vs. the indicated group. Cont, control; HG, high glucose; DAPA, dapagliflozin; siCON, control siRNA; and siSGLT2, SGLT2 siRNA.

Article Snippet: The SGLT2 inhibitor dapagliflozin (DAPA, AstraZeneca, Cambridge, UK) was used at a final concentration of 10–100 μM for 48 h.

Techniques: Inhibition, Incubation, Western Blot, Transfection, Control, Knockdown, Standard Deviation

Journal: Cells

Article Title: Dapagliflozin Restores Impaired Autophagy and Suppresses Inflammation in High Glucose-Treated HK-2 Cells

doi: 10.3390/cells10061457

Figure Lengend Snippet: SGLT2 inhibition suppressed HG-induced inflammatory alterations. ( A ) HK-2 cells were incubated with Cont, HG (30 mM), or HG (30 mM) dapagliflozin (20 µM) for 48 h, and p-AMPK, AMPK, IL-1β, and TNFα were assessed by western blotting. ( B ) Densitometric evaluation of ( A ) (n = 3). ( C ) Immunofluorescence of IL-6 in HK-2 cells cultured with Cont, HG (30 mM), or HG (30 mM) dapagliflozin (20 µM) for 48 h. ( D ) HK-2 cells were transfected with SGLT2 siRNA (200 nM) or control siRNA for 6 h and then incubated with Cont or HG (30 mM) medium for 48 h. P-AMPK, AMPK, IL-1β, and TNFα were assessed by western blotting. ( E ) Densitometric evaluation of ( D ) (n = 3). ( F ) Immunofluorescence of IL-6 in HK-2 cells transfected with SGLT2 siRNA (200 nM) or control siRNA for 6 h and then treated with Cont or HG (30 mM) medium for 48 h. All data represent the mean ± standard deviation (SD). * p < 0.05 vs. the indicated group, ** p < 0.01 vs. the indicated group (n = 3). Cont, control; HG, high glucose; DAPA, dapagliflozin; siCON, control siRNA; siSGLT2; and SGLT2 siRNA.

Article Snippet: The SGLT2 inhibitor dapagliflozin (DAPA, AstraZeneca, Cambridge, UK) was used at a final concentration of 10–100 μM for 48 h.

Techniques: Inhibition, Incubation, Western Blot, Immunofluorescence, Cell Culture, Transfection, Control, Standard Deviation

Journal: Cells

Article Title: Dapagliflozin Restores Impaired Autophagy and Suppresses Inflammation in High Glucose-Treated HK-2 Cells

doi: 10.3390/cells10061457

Figure Lengend Snippet: SGLT2 inhibition suppressed HG-induced nuclear localization of NF-κB p65. ( A ) HK-2 cells were incubated with Cont, HG (30 mM), or HG (30 mM) and dapagliflozin (20 µM) for 48 h, and the levels of NF-κB p65 in nucleoprotein extracts and NLRP3, IL-1β, TNFα in cytoplasmic protein extracts were assessed by western blotting. ( B ) Densitometric evaluation of ( A ) (n = 3). ( C ) HK-2 cells were transfected with SGLT2 siRNA (200 nM) or control siRNA for 6 h and then incubated with Cont or HG (30 mM) medium for 48 h. The levels of NF-κB p65 in nucleoprotein extracts and NLRP3, IL-1β, and TNFα in cytoplasmic protein extracts were assessed by western blotting. ( D ) Densitometric evaluation of ( C ) (n = 3). All data represent means ± standard deviation (SD). * p < 0.05 vs. the indicated group, ** p < 0.01 vs. the indicated group (n = 3). Cont, control; HG, high glucose; DAPA, dapagliflozin; siCON, control siRNA; siSGLT2, SGLT2 siRNA; NE, nucleoprotein extracts; and CE, cytoplasmic protein extracts.

Article Snippet: The SGLT2 inhibitor dapagliflozin (DAPA, AstraZeneca, Cambridge, UK) was used at a final concentration of 10–100 μM for 48 h.

Techniques: Inhibition, Incubation, Western Blot, Transfection, Control, Standard Deviation

Journal: Cells

Article Title: Dapagliflozin Restores Impaired Autophagy and Suppresses Inflammation in High Glucose-Treated HK-2 Cells

doi: 10.3390/cells10061457

Figure Lengend Snippet: The mechanism of SGLT2 inhibition in diabetic renal proximal tubular cells. High glucose-induced decrease in AMPK activity is involved in (1) mTOR activation and autophagic flux reduction and (2) inflammatory alterations (inflammasome activation and increased TNFα and IL-6 expression) due to NF-κB (p65) pathway activation. Dapagliflozin attenuates the alterations caused by high glucose.

Article Snippet: The SGLT2 inhibitor dapagliflozin (DAPA, AstraZeneca, Cambridge, UK) was used at a final concentration of 10–100 μM for 48 h.

Techniques: Inhibition, Activity Assay, Activation Assay, Expressing

Journal: RSC Advances

Article Title: SGLT inhibitors as antidiabetic agents: a comprehensive review

doi: 10.1039/c9ra08706k

Figure Lengend Snippet: Genes, substrates and tissue distribution of SGLT proteins 42

Article Snippet: 5 , WO2010048358A2 (US20100167988A1 ) , Auspex Pharmaceutical, Inc. , 2010 , Ethoxyphenylmethyl inhibitors of SGLT2 ( ) , United States.

Techniques:

Journal: RSC Advances

Article Title: SGLT inhibitors as antidiabetic agents: a comprehensive review

doi: 10.1039/c9ra08706k

Figure Lengend Snippet: Chemical classification of SGLT1/SGLT2 inhibitors.

Article Snippet: 5 , WO2010048358A2 (US20100167988A1 ) , Auspex Pharmaceutical, Inc. , 2010 , Ethoxyphenylmethyl inhibitors of SGLT2 ( ) , United States.

Techniques:

Journal: RSC Advances

Article Title: SGLT inhibitors as antidiabetic agents: a comprehensive review

doi: 10.1039/c9ra08706k

Figure Lengend Snippet: In vitro evaluation results of compounds 33–37 against SGLT2 ( ref. 99 )

Article Snippet: 5 , WO2010048358A2 (US20100167988A1 ) , Auspex Pharmaceutical, Inc. , 2010 , Ethoxyphenylmethyl inhibitors of SGLT2 ( ) , United States.

Techniques: In Vitro

Journal: RSC Advances

Article Title: SGLT inhibitors as antidiabetic agents: a comprehensive review

doi: 10.1039/c9ra08706k

Figure Lengend Snippet: Patents on SGLT inhibitors

Article Snippet: 5 , WO2010048358A2 (US20100167988A1 ) , Auspex Pharmaceutical, Inc. , 2010 , Ethoxyphenylmethyl inhibitors of SGLT2 ( ) , United States.

Techniques: Activity Assay

Journal: RSC Advances

Article Title: SGLT inhibitors as antidiabetic agents: a comprehensive review

doi: 10.1039/c9ra08706k

Figure Lengend Snippet: Genes, substrates and tissue distribution of SGLT proteins 42

Article Snippet: 3 , WO2014159151A1 (US9573959B2, EP3466431A1 , EP2968375B1) , MSD International GmbH , 2014 , Methods for preparing SGLT2 inhibitors , United States, Europe.

Techniques:

Journal: RSC Advances

Article Title: SGLT inhibitors as antidiabetic agents: a comprehensive review

doi: 10.1039/c9ra08706k

Figure Lengend Snippet: Chemical classification of SGLT1/SGLT2 inhibitors.

Article Snippet: 3 , WO2014159151A1 (US9573959B2, EP3466431A1 , EP2968375B1) , MSD International GmbH , 2014 , Methods for preparing SGLT2 inhibitors , United States, Europe.

Techniques:

Journal: RSC Advances

Article Title: SGLT inhibitors as antidiabetic agents: a comprehensive review

doi: 10.1039/c9ra08706k

Figure Lengend Snippet: In vitro evaluation results of compounds 33–37 against SGLT2 ( ref. 99 )

Article Snippet: 3 , WO2014159151A1 (US9573959B2, EP3466431A1 , EP2968375B1) , MSD International GmbH , 2014 , Methods for preparing SGLT2 inhibitors , United States, Europe.

Techniques: In Vitro

Journal: RSC Advances

Article Title: SGLT inhibitors as antidiabetic agents: a comprehensive review

doi: 10.1039/c9ra08706k

Figure Lengend Snippet: Patents on SGLT inhibitors

Article Snippet: 3 , WO2014159151A1 (US9573959B2, EP3466431A1 , EP2968375B1) , MSD International GmbH , 2014 , Methods for preparing SGLT2 inhibitors , United States, Europe.

Techniques: Activity Assay