The blood samples collected from the experimental animal were allowed to stand and centrifuged at 1000× g for 15 min to obtain serum. The mouse IL-1β and IL-6 ELISA Kit (CUSABIO, Wuhan, China) were used to determine the levels of IL-1β and IL-6, respectively.
After treating bEnd.3 cells with the corresponding drugs, the supernatant was collected for the measurement of IL-1β (CUSABIO, Wuhan, China). In addition, the treated bEnd.3 cells were washed with PBS and collected with cell lysis buffer. After centrifuging at 12,000× g for 15 min at 4 °C, the supernatant was collected for the measurement of sAC, cAMP, and CaMKII according to the protocol of sAC ELISA Kit, cAMP ELISA Kit (Elabscience, Wuhan, China) and CaMKII ELISA Kit (Meimian, Yancheng, China). The quantification of succinate was measured by succinate colorimetric assay kit (Sigma, MAK184, St. Louis, MO, USA) The activity of CaN and SDH were detected according to the calcineurin assay kit (Jiancheng, A068-1-1, Nanjing, China) and succinate dehydrogenase assay kit (Jiancheng, A022-1-1, Nanjing, China).
After treating bEnd.3 cells with the corresponding drugs, the supernatant was collected for the measurement of IL-1β (CUSABIO, Wuhan, China). In addition, the treated bEnd.3 cells were washed with PBS and collected with cell lysis buffer. After centrifuging at 12,000× g for 15 min at 4 °C, the supernatant was collected for the measurement of sAC, cAMP, and CaMKII according to the protocol of sAC ELISA Kit, cAMP ELISA Kit (Elabscience, Wuhan, China) and CaMKII ELISA Kit (Meimian, Yancheng, China). The quantification of succinate was measured by succinate colorimetric assay kit (Sigma, MAK184, St. Louis, MO, USA) The activity of CaN and SDH were detected according to the calcineurin assay kit (Jiancheng, A068-1-1, Nanjing, China) and succinate dehydrogenase assay kit (Jiancheng, A022-1-1, Nanjing, China).