Effect and mechanism of FAM134B-mediated ER-phagy on cerebral ischemia-reperfusion injury in mice
Objective To observe the effects of ER-phagy mediated by family with sequence similarity 134 member B(FAM134B)on cerebral ischemia-reperfusion(IR)injury in mice and to explore its mechanism.Methods Male C57BL/6 mice were randomly divided into the sham operation group(Sham group),cerebral IR group(IR group),and ce-rebral IR group with silenced FAM134B group(IR+siFAM134B group),with 6 mice in each group.Mice in the IR+si-FAM134B group received stereotactic injection in the brain of FAM134B silencing lentivirus,while mice in the Sham group and IR group were normally fed without lentivirus transfection.Seven days after transfection,mice in the IR group and IR+siFAM134B group were subjected to middle cerebral artery occlusion using thread occlusion method,while in the Sham group,we only isolated the middle cerebral artery without inserting the thread.After 24 h of reperfusion,HE and Nissl staining were used to observe brain damage in each group.Immunofluorescence was employed to observe apoptosis rate in the brain tissues,ELISA was used to detect the levels of oxidative stress indices[malondialdehyde(MDA),super-oxide dismutase(SOD),and catalase(CAT)],and Western blotting was utilized to detect the ER-phagy-related proteins(FAM134B,LC3B,Beclin-1,and p62).Results In the Sham group,no obvious pathological changes and apoptotic cells were seen in neuronal cells.In the IR group,brain tissues exhibited abnormal pathology,with irregular neuronal mor-phology,shrunken or absent nuclei,and a large number of apoptotic cells.While the IR+siFAM134B group showed re-duced pathological damage and fewer apoptotic cells in comparison with those of the IR group.Compared with the Sham group,the activity of SOD and CAT decreased,while MDA content increased in the IR group(all P<0.05).Compared with the IR group,the levels of SOD,CAT increased,whereas MDA content decreased in IR+siFAM134B group(all P<0.05).Compared with the Sham group,IR group had decreased level of P62 and increased levels of FAM134B,LC3B,and Beclin-1(all P<0.05).Compared with the IR group,the level of P62 increased,whereas FAM134B,LC3B and Be-clin-1 decreased in the IR+siFAM134B group(all P<0.05).Conclusion Inhibiting ER-phagy mediated by FAM134B can alleviate the cerebral IR injury in mice,and the mechanism may be related to reducing oxidative stress and inhibiting apoptosis in the brain tissues.
ER-phagysequence similarity 134 member Bcerebral ischemia reperfusion injuryoxidative stressapoptosis
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维普
