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For short, an equal amount of proteins was subjected to 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). After transferring the proteins onto polyvinylidene difluoride (PVDF) membranes, the membranes were washed with TTBS for 5 min and then incubated with skim milk powder solution for 1 h. Primary antibody against NF-��B subunit p65 (1:500), p-I��B�� (1:500), I��B�� (1:500), p-ERK (1:500), ERK (1:500), p-JNK (1:500), JNK (1:500), p-p38 (1:500), p38 (1:500), ��-actin (1:1000), or Histone H3 (1:500) was added and the membranes were incubated at 4��C overnight. Following washing with TTBS, the membranes were incubated with HRP-conjugated IgG secondary antibodies selleck chemical (1:5000) for 45 min at 37��C. After washing, the blots were developed using Beyo ECL Plus reagent and scanned in the Gel Imaging System. The relative expression levels of the target proteins were calculated with Gel-Pro-Analyzer (Media Cybernetics, USA). Statistical Analysis PLX3397 All the data were expressed in the form of mean �� SD. Multiple comparisons were conducted by Duncan method using general liner model with a significant level of 0.05. All the statistical analysis and image manipulation were conducted using R language version 3.2.1 (R Foundation for Statistical Computing). Results Administration of GP Alleviated the Impairments on the Cardiac Function and Structure Induced by I/R Injury in Rats Detail data of LVESP, LVEDP, and FS detection was shown in Table ?Table11. I/R injury caused significant reduction of LVESP and FS, but the value of LVEDP was increased post I/R injury induction. Administration of GP significantly ALOX15 alleviated the impairments on cardiac function due to I/R injury. For LVEDP and LVESP, average levels in I/R + GPM and I/R + GPH groups were significantly higher than those in I/R group (P