Ginsenoside rg1

The LYG formula contains a 1:2 ratio of Ginseng and Radix Rehmanniae Preparata. The formula was extracted by water, and its concentration was determined by the amount of the crude drug in solution. The Ginseng and Radix Rehmanniae Preparata were provided by Kangmei Pharmaceutical (Guangzhou, China), and the LYG was produced according to the detailed specifications of Jing Yue’s Complete Work. Ginsenoside Rg1 (Lot: A0503AS) and catalpol (Lot: 50728AS) were provided by Meilunbio (Dalian, China). All drugs were dissolved in dimethylsulfoxide (DMSO) and were diluted in E. coli OP50 solution when the drugs were added during the preparation of agar plates. Detailed information about the drugs used in the different groups is shown in Table 1.Table 1

The components, dose proportions, and original concentrations of the drugs in the different groups

Groups and drugs	Components	Dose proportion	Original concentration (mg/ml)a
LYG	Ginseng and Radix Rehmanniae Preparata	1:2	1 mg/ml
Rg1	Ginsenoside Rg1	–	150 µg/ml
Catalpol	Catalpol	–	150 µg/ml
Rg1 + Catalpol	Ginsenoside Rg and catalpol	1:1	150 µg/ml
Control	DMSO	–	Equal to the other groups

^aThe concentrations of the drugs used to treat N2 wild-type nematodes in this study were determined by a dose–effect curve and were chosen based on the dose with the greatest effect on the lifespan of N2 wild-type nematodes; DMSO, dimethylsulfoxide

Ginsenoside Rg1, Rg3, Rg5, Rb1, Rh2, CK (purity > 98% as determined by HPLC) and methotrexate were purchased from Dalian Meilun Biotech Co., Ltd. (Dalian, China). 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide (MTT), tumor necrosis factor (TNF-α), lipopolysaccharide (LPS), Type IV Collagenase were provided by Sigma-Aldrich Company (St. Louis, MO, USA). Propidium iodide (PI) and Annexin V-FITC were acquired from BioVision Inc. (Milpitas, CA, USA). Complete Freund’s adjuvant (CFA) and Type II bovine collagen were obtained from Chondrex, Inc. (Redmond, WA, USA). Mouse antibody CD45-Percp-Cyanine5.5, CD4-FITC, CD8-PE-Cy7, F4/80-FITC, CD86-PE were purchased from BioLegend Co, Ltd. (San Diego, CA, USA). TNF-α and the IL-6 ELISA Kit were gained from Thermo Fisher Scientific (Shanghai, China).