Fetal bovine serum (fbs)

YD sample (batch no. 20201045) was purchased from Guizhou Bailing Group Pharmaceutical Co., Ltd. (Guiyang, China). Seven intermediates under the preparation procedure of YD [10 ], including extract of Ginkgo Folium (GF), ethanol extract of Salviae Miltiorrhizae Radix et Rhizoma (ESM), water extract of Salviae Miltiorrhizae Radix et Rhizoma (WSM), mixed extract of Notoginseng Radix et Rhizoma, Crateagi Fructus, and Gynostemma pentaphyllum (Mix), water extract of Erigerontis Herba (EH), Borneolum (BN), oil of Allii Sativi Bulbus (AS) were offered from Guizhou Bailing Group Pharmaceutical Co., Ltd. (Guiyang, China).
The reference standard of scutellarin, ginsenoside Rd, salvianolic acid B, kaempferol-3-O-2ʺ-(6ʺ-p-coumaroyl) glucosyl rhamnoside, and ginsenoside Rb1 were purchased from Chengdu Must Bio-technology Co., Ltd (Sichuan, China). Dexamethasone acetate (DXM) and propafenone hydrochloride (PPH) were purchased from Macklin Co. (Shanghai, China). The purity of all reference standards was not less than 98%. Silica gel (5 μm, 100 Å) was purchased from Bonna-Agela Co. (Tianjing, China) and was activated at 105 ℃ for 120 min before use.
The LC/MS-grade acetonitrile and HPLC-grade methanol were purchased from Merck (Darmstadt, Germany). Deionized water (18 MΩ·cm) was purified by a Milli-Q water purification system from Millipore (Bedford, MA, USA). The HPLC-grade ammonium acetate was purchased from ROE Scientific Inc. (USA). Dulbecco’s modified Eagle’s medium (DMEM) and phosphate buffered saline (PBS) were purchased from Nanjing Keygen Biotech Co. (Nanjing, China). Fetal bovine serum (FBS) and trypsin were purchased from Gibco Life Technology Co. (Australia).

Human monocytic THP-1 cells were maintained in culture in Roswell Park Memorial Institute medium (RPMI 1640, Invitrogen) culture medium containing 10 % of heat inactivated fetal bovine serum (Invitrogen) and supplemented with 10 mM Hepes (Gibco, #15630-056), 1 mM pyruvate (Gibco, #11360-039), 2.5 g/l D-glucose (Merck) and 50 pM ß-mercaptoethanol (Gibco; 31350–010). THP-1 monocytes are differentiated into macrophages by 24 h incubation with 150 nM phorbol 12-myristate 13-acetate (PMA, Sigma, P8139) followed by 24 h incubation in RPMI medium. Macrophages were polarized in M1 macrophages by incubation with 20 ng/ml of IFN-γ (R&D system, #285-IF) and 10 pg/ml of LPS (Sigma, #8630). Macrophage M2 polarization was obtained by incubation with 20 ng/ml of interleukin 4 (R&D Systems, #204-IL) and 20 ng/ml of interleukin 13 (R&D Systems, #213-ILB). HepG2 and A549 cells were respectively cultivated in Dulbecco’s modified Eagle's minimal essential medium (DMEM medium 1 g glucose/l) (Gibco) and Minimum Essential Medium Eagle medium (MEM) (Gibco), both containing 10 % fetal bovine serum. In the co-culture experiments, THP-1 monocytes were differentiated in 6 Transwell inserts (membrane pore size of 0.4 μm, Corning, #3450). Macrophages and HepG2 cells were co-cultured in CO₂ independent medium supplemented with 0.5 mM L-glutamine (Sigma, # G3126) and 3.75 g/l of D-glucose (Sigma, #50-99-7) for 16 h before being incubated with or without 50 μM etoposide (Sigma, #E1383) for 24 h. Macrophages and A549 cells were co-cultured in CO₂ independent medium supplemented with 0.5 mM L-glutamine and 2.5 g/l of D-glucose for 24 h before being incubated with or without 50 μM etoposide for 16 h. In the monoculture experiments, 0.8 x 10⁶ THP-1 monocytes were differentiated and polarized in 6 well plates. Next, they were incubated in CO₂ independent medium supplemented with 0.5 mM L-glutamine (Sigma, # G3126) and 3.75 g/l of D-glucose (Sigma, #50-99-7) for 16 h before being incubated with or without 50 μM etoposide (Sigma, #E1383) for 24 h.