miR-21 Regulates Immune Balance Mediated by Th17/Treg in Peripheral Blood of Septic Rats during the Early Phase through Apoptosis Pathway

Figure 7

The levels of caspase-3 and caspase-9 in lymphocytes were changed after transfection of miR-21 activator or inhibitor into lymphocytes of septic rats. ((a) and (b)) The relative expressions of caspase-3 and caspase-9 mRNA in eight-paired NC rat group, SP rat group, SP rat + miR21 mimic group, and SP rat + miR21 inhibitor group were determined by real-time quantitative PCR [t value (caspase-3: NC versus SP) = 25.960,  < 0.01; t value (caspase-3: miR-21 mimic versus SP) = 10.874,  < 0.01; t value (caspase-3: miR-21 inhibitor versus SP) = 5.934,  < 0.01; t value (caspase-9: NC versus SP) = 13.300,  < 0.01; t value (caspase-9: miR-21 mimic versus SP) = 3.711,  < 0.01; t value (caspase-9: miR-21 inhibitor versus SP) = 12.930,  < 0.01]. ((c) and (d)) The expressions of caspase-3 and caspase-9 protein in eight-paired NC rat group, SP rat group, SP rat + miR21 mimic group, and SP rat + miR21 inhibitor group were determined by immunofluorescence microscope. ((e)–(g)) In in vivo experiments, the protein levels of caspase-3 and caspase-9 in eight-paired NC rat group, SP rat group, SP rat + miR21 mimic group, and SP rat + miR21 inhibitor group were determined by Western blot [t value (caspase-3: NC versus SP) = 10.643,  < 0.01; t value (caspase-3: miR-21 mimic versus SP) = 4.089,  < 0.01; t value (caspase-3: miR-21 inhibitor versus SP) = 4.545,  < 0.01; t value (caspase-9: NC versus SP) = 7.345,  < 0.01; t value (caspase-9: miR-21 mimic versus SP) = 4.664,  < 0.01; t value (caspase-9: miR-21 inhibitor versus SP) = 2.681,  < 0.05]. ((h)–(j)) In in vitro culture, the protein levels of caspase-3 and caspase-9 in eight-paired NC rat group, SP rat group, SP rat + miR21 mimic group, and SP rat + miR21 inhibitor group were determined by Western blot. NC group: normal control group; SP group: sepsis group; SP rat + miR21 mimic group: sepsis group with miR-21 activator transfected into lymphocytes; SP rat + miR21 inhibitor group: sepsis group with miR-21 inhibitor transfected into lymphocytes.  < 0.01.

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