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@#Objective To explore the protective effects of Acacetin on mouse microglial cells injured by oxygen-glucose deprivation/reperfusion (OGD/R) based on autophagy/ROS/NLRP3 inflammasomes signaling pathway.Methods Mouse microglial cells (BV2) were divided into normal control group,OGD model group and OGD+Acacetin group (10 μmol).After hypoxia for 6 hours and reoxygenation for 24 hours,the viability of BV2 cells was detected by 4-methylazolazolium blue (MTT) method.The death rate of cell was measured by lactate dehydrogenase (LDH) method.Reactive oxygen species (ROS) assay kit was used to measure the intracellular ROS level.The expression of LC3-Ⅱ,LC3-Ⅱ/LC3-Ⅰ,beclin-1,NLRP3,caspase-1 and IL-1β were analyzed by western blot.Results Acacetin increased the survival rate of microglia cells,and reduced the release of LDH and production of ROS.Acacetin also increased the expression of LC3-Ⅱ,and further to decrease the expression of NLRP3,caspase-1 and IL-1 β.Conclusions Acacetin protected against cerebral ischemia-reperfusion injury by attenuating the injury of microglia after OGD/R.Its protective mechanism may be related to inhibiting the production of ROS,activating autophagy,and inhibiting the NLRP3 inflammasome.
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Objective To investigate the effects of of inhibition of AMP-activated protein kinase (AMPK) on cytochrome c (CytC) expression in the cortex and hippocampus after focal cerebral ischemia-reperfusion in mice. Methods Thirty-six male C57BL/6 mice were randomly divided into 3 groups: a sham operation group, an ischemia-reperfusion group, and a compound C group (n = 12 in each group). The mice of the compound C group were intraperitonealy injected an AMPK specific inhibitor compound C (20 mg/kg) at the time of ischemia. A model of middle cerebral artery occlusion was induced by a modified suture method. After 24 h of reperfusion, Western blot was used to detect the expression levels of AMPK, phosphorylated-AMPK (p-AMPK), and cytoplasm CytC in the cortex and hippocampus in the ischemic side. Results p-AMPK/MPK levels in the cortex in the sham operation group, ischemia-reperfusion group, and compound C group were 0. 701 ± 0. 197, 1. 408 ± 0. 322, and 0. 930 ± 0. 229, respectively (F = 12. 000, P =0. 001); p-AMPK/MPK levels in the hippocampus were 0. 685 ± 0. 228, 1. 507 ± 0. 418, and 0. 964 ± 0. 378, respectively ( F = 8. 530, P = 0. 003 ); p-AMPK/AMPK levels both in the cortex ( P < 0. 001 ) and hippocampus (P = 0. 001) in the ischemia-reperfusion group were significantly higher than those in the sham operation group, p-AMPK/AMPK levels both in the cortex (P = 0. 005) and hippocampus (P = 0. 017) in the compound C group were significantly lower than those in the ischemia-reperfusion group. CytC levels in the cortex in the sham operation group, ischemia-reperfusion group, and compound C group were 0. 496 ±0. 278, 1. 461 ± 0. 321, and 1. 018 ± 0. 175, respectively (F = 19. 915, P < 0. 001); CytC levels in the hippocampus were 0. 511 ± 0. 257, 1. 610 ± 0. 441, and 0. 921 ± 0. 228 (F = 17. 795, P < 0. 001); CytC levels both in the cortex (P < 0. 001) and hippocampus (P < 0. 001) in the ischemia-reperfusion group were significantly higher than those in the sham operation group, while CytC levels both in the cortex (P = 0. 011) and hippocampus (P = 0. 002) in the compound C group were significantly lower than those in the ischemia-reperfusion group. Conclusion Inhibition of the AMPK may down-regulate the cytoplasm CytC expression in the cortex and hippocampus after cerebral ischemia-reperfusion in mice.