For the establishment of an Ox-LDL-induced apoptosis model and measurement of Gypenoside XVII’s protective effect, the HUVECs are seeded in 96-well plates at a density of 10⁵ cells per mL and grown for 24 h. Then, the HUVECs are pretreated with Gypenoside XVII (6.25, 12, 25, 50, 100 μg/mL) for 12 h in serum-free endothelial cell basal medium, followed by incubation with Ox-LDL (100 μg/mL, 24 h) which does not have Gypenoside XVII. After 24 h, the treated HUVECs are incubated with 5 mg/mL MTT in fresh medium for an additional 4 h. Absorbance is measured at 570 nm using a plate reader^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[1]
A total of 42 ApoE^-/- mice and 12 male C57BL/6J mice with the same age and body weight (approximately 20 g in body weight and 6 weeks old) are randomly divided into 4 groups (12 mice for each group) and orally administered the following once a: C57 control group (vehicle; 0.5% sodium carboxymethylcellulose, CMC-Na); ApoE^-/- model group (vehicle; 0.5% CMC-Na); ApoE^-/-+Gypenoside XVII group (Gypenoside XVII, 50 mg/kg via i.g. ); ApoE^-/-+probucol group (Probucol, 2 mg/kg via i.g.). Probucol is an antioxidant drug used as a positive control. After two weeks of acclimatization, all the mice are fed a high-fat diet including 0.3% cholesterol and 20% pork fat for 10 weeks. They are maintained in pathogen-free conditions at approximately 22±1°C on a 12 h light-dark cycle with free access to food and water. The body weights are determined every two weeks. After 10 weeks of the treatments, all animals are anesthetized with pentobarbital sodium and killed after being deprived of food overnight. Serum is immediately separated from blood samples by centrifugation at 3600 rpm for 15 min, and the tissue samples (heart and aorta) are rapidly removed and frozen in -8°C.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Yang K, et al. Gypenoside XVII Prevents Atherosclerosis by Attenuating Endothelial Apoptosis and Oxidative Stress: Insight into the ERα-Mediated PI3K/Akt Pathway. Int J Mol Sci. 2017 Feb 9;18(2). pii: E77.  [Content Brief]