Glucometer

In the 6th week of the mouse study, mice were fasted for 16 h and then weighted. Blood was taken from the tail vein for measurement of initial blood glucose level using a glucometer (Roche, Shanghai, China). After that, mice were intraperitoneally injected with glucose (Beyotime, Shanghai, China) at a dose of 1.5 g/kg⋅ BW. glucose was measured in whole blood from the tail vein at 0, 30, 60, 90, and 120 min using a glucometer.
Mice were fasted for 6 h before ITT. Blood was taken from the tail vein for measurement of initial blood glucose level. Mice were intraperitoneally injected with 1 IU/kg⋅ BW recombinant human insulin (Beyotime, Shanghai, China) and the glucose levels in whole blood from the tail vein were measured at 0, 30, 60, 90, and 120 min using Roche glucometer.

For the fasting blood glucose, glucose tolerance assays, and insulin tolerance exam as described by Amir [21 (link)]. After 12 weeks of treatment, fasted mice (16 h, paper bedding) by monitoring glucose levels after a glucose bolus (1 g/kg of body weight (BW)) or insulin (0.5 U/kg BW) by intraperitoneal (IP) injection. The exam of glucose was carried from fasted (4–16 h) or re-fed animals (15 min to 1 h). Re-feeding was conducted by injecting a bolus of glucose (1 g/kg of BW) IP as mentioned above. The first drop of blood was thrown away and then the second drop of blood was detected by the glucometer (Roche Diagnostics, Mannheim, Germany). We collected the tail blood samples at 0, 15, 30, 60, and 120 min after glucose loading and detected the blood glucose value by the glucometer (Roche Diagnostics, Mannheim, Germany).

After intraperitoneal injection of acacetin, oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were respectively carried out in line with a previous description [16 ].
For OGTT, the mice with miR-23b-3p mimic and NEU1 overexpression were fasted overnight for 12 h before each experiment. A tail cut (1–2 mm) was performed with a sterilized scissor (FS001; Beyotime, Shanghai, China), and around 30 μL of blood sample was collected using fresh capillary tube (15401–560, VWR, Atlanta, GA) for quantifying basal blood glucose level (= time point 0) with a glucometer (Roche Diagnostics, Switzerland). 15 min (min) after oral administration of D-glucose (2 g/kg mice body weight, Sigma-Aldrich), about 30 μL blood was collected to measure blood glucose level.
For ITT, mice with miR-23b-3p mimic and NEU1 overexpression were fasted overnight for 12 h before each experiment, and basal blood glucose level was measured at 0 min. Insulin (0.5 U/kg mice body weight, I5500, Sigma-Aldrich) was intraperitoneally injected into the mice, and whole blood glucose level was measured by a glucometer (Roche Diagnostics, Switzerland) after 15 min.