Animal experiments
Male KM mice (7-8 weeks, 18-22 g, RRID:MGI:5651867) were purchased from
the Laboratory Animal Centre of Sun Yat-sen University (Guangzhou,
China) and male Zucker Diabetic Fatty (ZDF) rats including ZDF-Leprfa/fa
rats and ZDF-Leprfa/ctrl rats (7-8 weeks, 220~240 g,
RRID:RGD_2314027, RRID:RGD_2311071) were purchased from Beijing Vital
River Laboratory Animal Technology (Beijing, China). All animal
procedures were approved by the Sun Yat-sen University Committee on
Ethics for the Use of Laboratory Animals in accordance with the Animal
Welfare Legislation of China. Animal studies are reported in compliance
with the ARRIVE guidelines.
KM mice were kept at 22 ± 1 °C on a 12-hour light/dark cycle with free
access to food and water. These mice were randomly divided into 6
groups, including lean group (without glucose administrated), vehicle
group, metformin group, Dapagliflozin group, AST group andCAG group, each group included 10 mice. Metformin (100 mg/kg),
Dapagliflozin (1 mg/kg), AST (5 mg/kg), CAG (5 mg/kg)
were dissolved in normal saline containing 1% CMC-Na and taken gavage
administration in mice. The lean and the vehicle groups were
administered the same volume. After acclimatization, oral glucose
tolerance test (OGTT) was performed. Briefly, mice were fasted for a
period of 14 h and then administrated with glucose (2 g/kg) orally. The
plasma glucose of 0, 15, 30, 60, and 120 min were detected by using a
blood glucose meter (Roche), respectively.
ZDF-Leprfa/ctrl rats were fed a chow diet provided by
the Animal Center of Sun Yat-sen University and
ZDF-Leprfa/fa rats were fed high fat diet 5008
(Labdiet, USA) for one month. Randomly selected the rats with blood
glucose over 11.1 mmol/L from ZDF-Leprfa/fa rats
before experiments. These ZDF rats were divided into 5 subgroups: lean
group (ZDF-Leprfa/ctrl, saline), vehicle group
(ZDF-Leprfa/fa, model), Dapagliflozin (1 mg/kg/d,p.o. ), CAG (1 and 5 mg/kg/d, p.o. ). The OGTT
experiment was preceded after the administration of CAG as
described above. The urinary glucose, urinary sodium, and urine efflux
were determined after 1- or 5-week treatment, respectively.