Milliplex map rat cytokine chemokine magnetic bead immunoassay kit

The homogenate tissue levels of inflammatory mediators, including TNF-α, IL-6, IL-4 and IL-10, were measured using a Milliplex MAP Rat Cytokine/Chemokine magnetic bead immunoassay kit (Millipore Corporation, Billerica, MA)[11 (link)]. The values were read with a MAGPIX Luminex xMAP instrument (Luminex Corp, Austin, TX) and analyzed with the MILLIPLEX Analysis software version 3 (Millipore Corporation, Billerica, MA)[11 (link)]. Blood samples (5 mL) were collected for centrifugation at 3000 rpm for 7 min at low temperature, and then, the supernatants were obtained for the alanine transaminase (ALT) and aspartate transaminase (AST) detection with an Automatic Biochemical Analyzer (AU5400, SIEMENS, Munich, Germany).

In the pharmacokinetic experiment, after administration of a single dose of SJD, a 0.5 mL blood sample was collected via the tail vein at 10, 20, and 40 min and 1, 2, 3, 4, 6, 8, and 12 h. After centrifugation at 3000 r/min for 7 min, the serum samples were stored at -80 °C for detection by HPLC-MS/MS. The rats were sacrificed 12 h after administration of SJD; pancreatic tissue samples were homogenized and the supernatants were obtained after centrifugation at 3000 r/min for 7 min and stored at -80 °C for detection.
In the pharmacodynamic experiment, the rats were sacrificed 12 h after administration of SJD. Blood samples (5 mL) were collected to obtain serum samples to measure amylase and lipase, using an Automatic Biochemical Analyzer (AU5400, SIEMENS, Munich, Germany), and to measure interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α levels using a Milliplex MAP Rat Cytokine/Chemokine magnetic bead immunoassay kit (Millipore Corporation, Billerica, MA, United States). Heart, lung, liver, spleen, pancreas, kidney, and intestine tissue samples were collected for pathological examination.