Ab213909

Plasma levels of Angiopoietin-2 (MANG20, R&D Systems, Minneapolis, MN, USA) and Endocan-1 (MBS039900, MyBioSource, San Diego, CA, USA) were measured as markers of endothelial dysfunction by ELISA according to the manufacturer’s protocol. Plasma, kidney, and lung IL-1β (RLB00, R&D Systems) and IL-18 (ab213909, Abcam, Boston, MA, USA) were quantified by ELISA according to the manufacturer’s protocol. Kidneys and lung lysates were prepared with the BioPlex Cell Lysis Kit (171304011, BioRad, Hercules, CA, USA) according to the manufacturer’s protocol.

The expressions of interleukin-18 (IL-18) (Abcam, USA, No. ab213909), IL-6 (Abcam, USA, No. ab234570), IL-1β (Abcam, USA, No. ab255730), tumor necrosis factor-α (TNF-α) (Abcam, USA, No. ab236712), and reactive oxygen species (ROS; BlueGene Biotech, Shanghai, China, No. 21175D-96) in serum samples were quantified according to the instructions of the ELISA kits. In brief, uterus and fallopian tube tissues were placed in precooled phosphate-buffered saline (PBS) and homogenized. The supernatant was collected by centrifugation at 4000 r/min for 10 min at 4°C. The BCA assay kit (Thermo Fisher Scientific, USA) was used to determine the protein content of the supernatant. 15 ng/μL of the protein samples was used for incubation with corresponding antibodies. The absorbance was measured at 450 nm wavelength and was estimated using an enzyme-linked immune monitor (Thermo Fisher Scientific, USA). The concentration of these proteins in the sample was calculated from the standard curve.

The levels of interleukin-1β (IL-1β) and IL-18 in HMC3 cell lysates were measured by ELISA using a human IL-1β ELISA kit (ab214025, Abcam) and a human IL-18 ELISA kit (ab215539, Abcam) according to the manufacturer's instructions. IL-1β and IL-18 levels in rat serum were detected by a rat IL-1β ELISA kit (ab100768, Abcam) and a rat IL-18 ELISA kit (ab213909, Abcam), respectively.