Neferine

Cell experiment:

Cardiac fibroblasts (CFs) are isolated from neonatal mouse ventricular tissues. After starvation in serum-free medium for 24 h, CFs are incubated in DMEM containing 5.6 mM glucose (normal glucose; NG), 30 mM D-glucose (HG), 30 mM D-glucose plus 1 μM Neferine, 30 mM D-glucose plus 2 μM Neferine, 30 mM D-glucose plus 5 μM Neferine, and 5.6 mM glucose plus 27.5 mM mannose. Cells are harvested at 24 h, 48 h, and 72h. Cell proliferation is measured with the Cell Counting Kit-8 (CCK-8) and the Cell-LightTM EdU assay[2].

Animal experiment:

Mice[2]Eight-week-old C57BL/6J male mice are used. Diabetes is induced by intraperitoneal injection of Streptozotocin dissolved in citrate buffer (pH 4.5) at 60 mg/kg body weight for five consecutive days. Control mice are injected with citrate buffer only. Whole blood glucose in mouse tail blood is detected with an Accu-Check Active glucometer. Mice with blood glucose concentrations higher than 18 mM are considered as diabetic animals and used in this study. The animals are randomly divided into four groups of eight animals each. Diabetic mice are divided into three groups: group 1, the diabetic control group (DM); group 2, which receive Neferine at a dose of 60 mg/kg/day (DM-NL); and group 3, which receive Neferine at a dose of 120 mg/kg/day (DM-NH). Neferine is administered twice per day by intragastric gavage for 12 weeks. Equivalent volumes of normal sodium are administered to the normal and DM control groups by gavage. Mice are anaesthetized and sacrificed at the end of the 12-week treatment[2].

References:

[1]. Baskaran R, et al. Neferine prevents NF-κB translocation and protects muscle cells from oxidative stress and apoptosis induced by hypoxia. Biofactors. 2016 Jul 8;42(4):407-17.
[2]. Liu X, et al. Neferine inhibits proliferation and collagen synthesis induced by high glucose in cardiac fibroblasts and reduces cardiac fibrosis in diabetic mice. Oncotarget. 2016 Sep 20;7(38):61703-61715.