Accu chek performa glucometer

Plasma total cholesterol (TC), TG and HDL-c were determined before and after the experimental protocol by enzymatic colorimetric kit (Labtest, Brazil) and glucose by Accu-Chek^® Performa glucometer (Roche, Brazil). CETP activity was determined by the transfer of ¹⁴C-cholesteryl oleate from human HDL to human VLDL and LDL, after incubation with plasma from CETP-tg mice (Escolà-Gil et al., 2001 (link)).

Mice were separated into 8 groups of 5 mice each and received an intraperitoneal (IP) injection with the following treatments: Group 1 control, saline solution (4 mL/kg); Group 2, glibenclamide as a positive control (10 mg/kg); Groups 3, 4, and 5, different doses of aqueous alkaloid-free extract (125, 250, and 500 mg/kg, respectively); Group 6, aqueous fraction (250 mg/kg); Group 7, organic fraction (250 mg/kg); and Group 8, precipitate (250 mg/kg). Blood samples were collected from the tail vein before (basal glycemia, t=0) and 2, 4, and 6 h after treatments. Glucose quantification was carried out using the glucose oxidase method with an Accu-Chek™ Performa glucometer (Roche Diagnostics, USA).

Venous blood samples were collected immediately after exercise, and blood analysis was performed using whole blood and serum. To collect the serum, the venous blood samples were allowed to clot at 24–25 °C for 30 min, centrifuged at 2000× g for 15 min at 4 °C, and transferred to new tubes before being stored at −80 °C. Whole blood was used to measure the concentration of blood glucose (ACCU CHEK Performa Glucometer, Roche, Diagnostics, Penzberg, Germany), lactate (Lactate Pro2, LT-1730, ARKRAY, Kyoto, Japan), and TG (Standard LipidoCare Strips–Lipid Profile, 02LA10G, SD Biosensor, Suwon, Korea). Serum was used in ELISA kits for the analysis of the concentration of glycerol (EGLY-200, BioAssay System, Hayward, CA, USA) and FFAs (K612-100, BioVision, Milpitas, CA, USA).

The body weight of mice and glycemic events were monitored at regular intervals. Mice were fasted for 4 h, and glucose levels (mmol/L) were measured using an Accu-Chek Performa Glucometer (Roche Diagnostics, Mannheim, Germany). Fasting insulin levels in serum were measured using the Mouse Insulin ELISA Kit (CSB E05071m, Cusabio biotech). Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) was quantified as blood glucose level multiplied by serum insulin level divided by 22.5. Soluble galectin 3 levels in serum were measured using the galectin 3 mouse ELISA Kit (ab203369, Abcam). After sacrifice, the amount of the epididymal visceral adipose tissue (VAT) was measured for each mouse. Serum concentrations of lipids were measured using the Olympus AU600 Chemistry Immuno Analyzer (Olympus, Tokyo, Japan).

Male wild type BKS mice (C57BLKS/J, the Jackson Laboratory stock number 000662) and BKS db mice (BKS.Cg^-Dock7^m+/+Leprdb/J, the Jackson Lab stock number 000642), aged six- to eight-weeks, were supplied by the Model Animal Research Center of Nanjing University (MARC). The mice were provided with individually ventilated cages in a specific pathogen free (SPF) animal rooms and were maintained temperature (24 ± 2 °C), humidity (60–80%), under 12 h light/dark cycle. All mice were fed sterile water and a normal chow diet ad libitum.
The mice for this study were administered according to the guidelines of protocol HAIFAJIZI-2013-3 (approval date: 9 December 2013) approved by the Institute of Oceanology committees for care and use of laboratory animals. A week after the mice adapted to the environment, all diabetic BKS db mice which accord with the experimental requirements were randomly divided into 3 groups: diabetic model group (db group), metformin-treated group (Met group, 100 mg.kg⁻¹.body⁻¹.day⁻¹), BDB-treated group (BDB group, 100 mg.kg⁻¹.body⁻¹.day⁻¹). Age-matched male wild type BKS mice were used as the normal control (BKS group). After 7 weeks, we measured the body weights, food intake, water intake and fasting glucose levels using an Accu-Chek Performa glucometer (Roche, Mannheim, Germany).

Alloxan monohydrate (Sigma Chemical Company, USA) was used to induce diabetes in mice and Glibenclamide (Hoechst Pharmaceuticals, Mumbai) was used as a standard hypoglycemic drug. Ethanol (BDH Ltd., England) and distilled water were used for extraction of the plant materials. ACCU CHEK Performa Glucometer (Roche Diagnostics India Pvt. Ltd., India) was used to measure the blood glucose level. For evaporating the solvents, BUCHI Rotavapour R-200, Switzerland and Lyophilizer (freeze dryer) (type: Heto power dry LL3000 Wag tech) was used. The following chemicals were used for phytochemical screening test: Chloroform and Ethyl acetate (ACS, Merck); Hydrochloric acid, Ferric sulphate, Lead acetate and Potassium ferrocyanide (BDH Ltd., England); Petroleum ether 60-80 °C (Labmerk Chemicals LTD India); Sulphuric acid (Farm Italia Carrloerba, Italy); Acetic anhydride and Methanol HPLC grade (Techno Pharmchem, Bahadurgarm, India); n-Hexane (Rathburn Chemicals Ltd., England); Acetonitrile (Sigma Aldrich, Germany) and Ferric chloride (FISHER Scientific Company, USA). All the chemicals were of analytical grades.

After 11 weeks of diet treatment and colon carcinogenesis induction, the mice were fasted for 6 h prior to receiving intraperitoneal injections of glucose (2 g/kg BW). Blood samples were collected from the tail vein before and at 15, 30, 60, and 90 min post-glucose injection. The levels of glucose were measured by Accu-Chek^® Performa glucometer (Roche^®, São Paulo, SP, Brazil). Differences in glycaemia before and during glucose administration were used to calculate the area under the curve (AUC).