电针上调Cezanne表达抑制NF-κB信号通路介导的大鼠脑缺血/再灌注炎性损伤
摘要
Objective To investigate the roles of electroacupuncture (EA) in the upregulation of zinc finger protein Cezanne and in its neuroprotective effect on inflammatory injury induced by focal cerebral ischemia/reperfusion (I/R) in rats. Methods SD rats were randomized into sham group, EA group, I/R group and I/R+EA group. Cezanne silencing lentivirus (LV-shCezanne) was injection into the cerebral cortex to study the possible effect of Cezanne silencing on I/R injury, and injection of blank lentivirus (vehicle) as the control. The I/R model was established by conventional method of middle cerebral artery occlusion with ischemia for 2 h followed by reperfusion for 6, 12, 24, 48 and 72 h and 7 d, respectively, and then EA was adopted to stimulate the Baihui acupoint and the Hegu and Taichong acupoints on the affected side. Garcia score, TTC staining, Western blotting, cell counting and immunofluorescence assay were performed in each group, so as to detect the neurological deficit score (NDS), infarct size, Cezanne expression in the ischemic cerebral cortex, number and distribution of Cezanne positive cells, and expression of nuclear factor kappa B (NF- κB) p65 in the nucleus and cytoplasm. Results Compared with the sham group, the protein level of Cezanne was increased in EA group at 6 and 48 h (P<0.05), and also in the I/R group at 12, 24, 48 and 72 h (P<0.05). While the level was further increased in the I/R+EA group at 12, 24, 72 and 7 d than the I/R group (P<0.05). The NDS was significantly lower in the I/R group than the sham group (P<0.05), whereas the number of Cezanne positive cells and the size of cerebral infarct were obviously larger (P<0.05). Compared with the I/R group, the I/R+EA group had higher NDS, more Cezanne positive cells, and smaller cerebral infarct size (P<0.05), and the ratios of both p-IκBα/IκBα and nucleus-p65 to cytoplasm-p65 were decreased in the MCAO/R+EA group (P<0.05). Immunofluorescence assay showed that Cezanne was mainly distributed in the