Penicillin and streptomycin

C166, a mouse vascular endothelial cell line used in this study, was purchased from Honsun Biological Technology Co., Ltd. (Shanghai, China). RAW264.7 cell line was provided by Professor Zhang Luo. C166 cells and RAW264.7 cells were cultured with DMEM (Hyclone^TM, USA) containing 10%FBS (Sigma, USA) and 1% antibiotics (penicillin and streptomycin; Biosharp, China) at 37 °C in a 5%CO₂ incubator.
CCK‐8 assay was used to determine endothelium viability (MCE, USA). Briefly, C166 cells were seeded in a 96‐well plate at 5 × 10³ cells·well⁻¹ and treated with LPS at different doses and time points. Then CCK‐8 was added into the culture medium for 3 h and the absorbance at 450 nm was determined by a microplate reader. Cell viability was calculated according to the manufacturer's protocols.

RAW264.7 macrophages were obtained from the Department of Pharmacology of Traditional Chinese Medicines, China Pharmaceutical University (Nanjing, China), and were cultured in Dulbecco’s modified essential medium (DMEM, Gibco) containing 10% fetal bovine serum (FBS, Sigma–Aldrich) and penicillin and streptomycin (Biosharp) at 37 °C in a humidified 5% CO₂ incubator (3111 type, Thermo Fisher Scientific). Cells were seeded in a 96-well culture plate at a density of 10 × 10⁵ cells/well and allowed to attach at 37 °C for 24 h [33 ]. Cells were treated with SGR extracts (final concentration, low (L), medium (M) and high (H) dose of 0.53, 1.06, and 2.13 mg/mL, respectively) and LPS (final concentration, 1 µg/mL) for an additional 24 h. The formazan crystals were dissolved in 150 µL of dimethyl sulfoxide (DMSO) for 15 min, and the absorbance was detected at 570 nm.

The SV40-transformed human embryonic kidney cell line HEK-293T, as well as HNSCC cell line Cal27, were purchased from Procell and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Biology industry, Shanghai, China) and 1% penicillin and streptomycin (Biosharp, Shanghai, China). The human head and neck squamous cell line UM1 was a generous gift from Prof. Anxun Wang (Department of Oral and Maxillofacial Surgery, First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China) [75 (link),76 (link),77 (link),78 (link)], and cultured in DMEM/F12 medium (Hyclone, UT, USA) supplemented with 5% FBS and 1% penicillin and streptomycin. Cell culture was carried out at 37 °C, 5% CO₂ and 95% humidity.