Kac1211

The concentration of secreted protein was assessed by commercial ELISA kits depending on the method proposed by the manufacturer. For the detection of inflammation factors, tumor necrosis factor-α (TNF-α, KHC3011, Invitrogen, CA, USA), interleukin-6 (IL-6, KAC1261, Invitrogen, CA, USA), interleukin-1β (IL-1β, KAC1211, Invitrogen, CA USA), and oxidative stress indexes, malonic dialdehyde (MDA, S0131, Beyotime, Shanghai, China), superoxide dismutase (SOD, S0109, Beyotime, Shanghai, China) and glutathione peroxidase (GSH-px, S0052, Beyotime, Shanghai, China) in culture medium, the culture supernatant of HMCs after different treatments were collected and centrifuged to remove the cell debris. The clear supernatant was used for ELISA based on manufacturer’s instructions. Subsequently, the optical density (OD) value was detected using a Synergy H1 microplate reader (Winooski, Vermont, USA). The standard curve was created based on the concentration and OD value of the standard molecule provided in the kit. Finally, the concentration of cytokines and oxidative molecules were calculated based on the standard curve.