Enzymelinked immunosorbent assay (ELISA)
Blood samples were collected from the tail vein of the rats, centrifuged at 3000 rpm for 10 min, and serum was collected. The concentration gradient of the standard was diluted according to the instructions of the IL-6 ELISA kit and the SP ELISA kit. IL-6 standard concentration dilution gradient: 160, 80, 40, 20, 10, 5 pg/mL; SP standard concentration dilution gradient: 8, 4, 2, 1, 0.5, 0.25 ng/mL. 50 μL of different concentrations of standards were added to each standard well, and 50 μL of the tested sample was added to the sample well to be tested. Except for blank wells, 100 μL of horseradish peroxidase labelled detection antibody was added to each well of standard wells and sample wells to be tested, gently shaken to mix evenly, and incubated at 37℃ for 1 h. After repeated washing five times, 50 μL of each substrate A and B were added to each well and incubated at 37 ° C in the dark for 15 min. Finally, 50μL of termination solution was added to each well, and the liquid changed from blue to yellow to terminate the reaction. The absorbance values of each well were measured at 450 nm using a microplate reader.