E el h0088c

SZ95 sebocytes were seeded in 24-well plates (1×10⁵ cells/well). After 48-h exposure to different concentrations of PM2.5 suspension, the supernatant was collected at 4°C, centrifuged for 20 min at 1,000 × g at 4°C, and the supernatant was collected and stored at −20°C for subsequent assays. Concentrations of IL-1α, IL-6 and IL-8 were determined using commercial ELISA kits (E-EL-H0088c, E-EL-H0102c and E-EL-H0048c; ElabScience Biotechnology Co., Ltd., Wuhan, China) according to the manufacturer's instructions.

The levels of TNF-α (KHC3011, Thermo, USA), IL-6 (EH2IL6, Thermo, USA), and IL-1 β (E-EL-H0088c, Elabscience, USA) in tissue lysates and cell culture medium were detected by using commercial ELISA assay kit following manufacturer's protocols.

MT-4 cells were inoculated in 6-well plates at a density of 1 × 10⁶ cells/well, incubated at 37°C with 5% CO₂ for 24 h. The supernatant was discarded and 1 mL/well of titrated HIV-1 virus supernatant was added and incubated for 48 h. High, medium and low concentrations of serum containing JACKP and positive control drug lentinan (800ug/mL) 2mL/well were added to each well, then incubated for 24 h. The cells and supernatant of each well were collected, washed with PBS solution and stored at −80°C for analysis. ELISA assay was performed according to the reagent manufacturer’s instructions to determine the level of inflammatory cytokine IL-1ɑ (E-EL-H0088c, Elabscience, China), IL-1β (EK101BHS-96, MULTI SCIENCES, China), IL-2 (EK102HS-96, MULTI SCIENCES, China), IL-6 (EK106HS-96, MULTI SCIENCES, China), IL-6ST (E-EL-H6015, Elabscience, China) and IL-10 (EK110HS-96, MULTI SCIENCES, China) in the cells.