Lipopolysaccaride lps

Notoginsenoside R₁ (R₁), ginsenoside Rb₁ (Rb₁), ginsenoside Rb₂ (Rb₂), ginsenoside Rb₃ (Rb₃), ginsenoside Rg₁ (Rg₁), ginsenoside Rg₂ (Rg₂), ginsenoside Rf (Rf), ginsenoside Ro (Ro), ginsenoside Rd (Rd), ginsenoside Re (Re), (20S)-ginsenoside Rg₃ ((20S)-Rg₃), (20R)-ginsenoside Rg₃ ((20R)-Rg₃), ginsenoside Rk₂ (Rk₂) (≥98% pure) were provided by the Shanghai Yuanye Biotechnology Co., ltd. (Shanghai, China). P. ginseng, A. macrocephala, P. cocos and G. uralensis applied in the study were purchased from Fusong Shenyuan Changbaishan Ginseng Technology Co., ltd. (Fusong, China).
Acetonitrile was purchased from Fisher Scientific (Waltham, MA, USA). Water was purified through a Milli-Q water purification system (Millipore, MA, USA). TNF-α, IL-1β, IL-6 and NO ELISA kits were provided by Jiangsu Mai Sha Industry Co., ltd (Yancheng, China). Dextran sulfate sodium (DSS) was obtained from MP Biomedicals (California, USA). Cell Counting Kit-8 (CCK-8) was obtained from Biosharp Life Science (Shanghai, China). Lipopolysaccaride (LPS) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Griess reagents were purchased from Promega (Madison, USA).

FLSs were isolated from synovial tissues. Briefly, the synovial tissues were sectioned and digested with 0.25% trypsin to isolate the synoviocytes at 37˚C for 30 min. Following culturing overnight in DMEM supplemented with 10% FBS as well as penicillin (100 IU/ml) and streptomycin (100 µg/ml) at 37˚C, the non-adherent cells were removed and the adherent cells were cultured in DMEM (Thermo Fisher Scientific, Inc.) containing 10% FBS and 1% mixture of penicillin and streptomycin at 37˚C in a humidified atmosphere containing 5% CO₂. Cells were collected and used for subsequent assays at passage 4-8. FLSs were stimulated with 10 µg/ml lipopolysaccaride (LPS; Sigma-Aldrich; Merck KGaA) for 24 h at 37˚C to establish the RA model.