Ginsenoside rb1

The chemical constitutions of PNS and PNE were quantitatively determined using a Waters ACQUITY-UPLC CLASS system (Waters Corp., USA) coupled with an ACQUITY UPLC BEH phenyl column (150 mm × 2.1 mm, 1.7 μm) maintained at 45°C. Elution was performed with a mobile phase of water (A) and Acetonitrile (B) under a gradient program: 0-10 min, 19% B; 10-15 min, 19-35% B; 15-20 min, 35-38% B. The flow rate was 0.4 mL/min, and the injection volume was 2 μL. The analytes were monitored at the UV wavelength of 205 nm. Prior to the next injection, the column was washed with 100% B for 2 min and then equilibrated with the initial mobile phase for 3 min. Notogisenoside R1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1, and ginsenoside Rd (the purities of all standards were higher than 98% by HPLC analysis) were purchased from Chengdu Pufei De Biotech Co., Ltd. (China). Acetonitrile was purchased from RCI Labscan Limited (Thailand) of HPLC grade. Milli-Q water was prepared using a Milli-Q system (Millipore, USA).

Ginsenoside rb1, neocryptotanshinone, caffeic acid and ligustilide (the purities of all standards were higher than 98% by high-performance liquid chromatography analysis) were purchased from Chengdu Pufeide Biotech Co., Ltd. (Chengdu, China).
TRIzol™ Reagent, Fetal bovine serum (FBS), 0.25% Trypsin–EDTA (w/v), Dulbecco’s modified eagle's medium (DMEM), penicillin–streptomycin (10,000 U/mL, P/S), and phosphate-buffered saline (PBS) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Human MTNR1A polyclonal antibody and GGT1 polyclonal antibody were purchased from CLOUD-CLONE CORP. (CCC, USA). Anti-rabbit IgG, HRP-linked antibody was purchased from Cell Signaling Technology (Danvers, MA, USA). β-actin was purchased from COHESION BIOSCIENCES (SUZHOU, CHINA).