中国药理学通报Issue(2)：216-222,7.DOI:10.3969/j.issn.1001-1978.2016.02.014
EGFR siRNA通过阻碍STAT3磷酸化抑制大鼠星形胶质细胞活化
EGFR siRNA inhibits activation of astrocytes derived from rats through blockade of STAT3 phosphorylation
摘要
Abstract
Aim To observe the expression of epider-mal growth factor receptor ( EGFR) in cerebral tissues around hematomas after intracerebral hemorrhage, and explore the effects of EGFR on activation of astrocytes derived from rats and the involved mechanisms. Meth-ods The specimens of cerebral tissues around hemo-tomas after intracerebral hemorrhage undergoing hemo-tomas removal operation were collected and then divid-ed into 4 groups according to the time of intracerebral hemorrhage: <1 d, 1 ~5 d, 6 ~10 d and >10 d groups. Each group included 20 cases. At the same time, 20 dropped brain tissues distant to hemorrhage in the operative process were collected as control group. Immunohistostaining and Western blot were used to measure the expression of EGFR. After isolation and culturing, the astrocytes of rat cortex were treated with culture solution ( control group) , CNTF that was used to activate astrocytes, scramble siRNA + CNTF and EGFR siRNA +CNTF for 24h, respectively. The ex-pression of glial fibrillary acidic protein ( GFAP) mR-NA was detected through fluorescence real-time quanti-tative PCR. In addition, the protein levels of GFAP, signal transducers and activators of transcription 3 ( STAT3 ) and phosphorylated STAT3 ( p-STAT3 ) were examined using Western blot. Results With the ex-tension of intracerebral hemorrhage time, positive sig-nal index and protein expression levels of EGFR gradu-ally elevated, reached the peak on 6 ~10d, and then decreased after 10 d. There was statistical difference ( P<0. 01 ) . The expression levels of GFAP mRNA and protein as well as p-STAT3 were significantly in-creased in cells treated with CNTF alone as compared to control group ( P <0. 01 ) , whereas these effects were almost completely reversed by EGFR siRNA transfection ( P <0. 01 ) . Additionally, there was no statistical difference in STAT3 protein levels among groups ( P >0. 05 ) . Conclusions EGFR expression is upregulated in the cerebral tissues around hemotomas after intracerebral hemorrhage. Gene silence of EGFR contributes to suppressing the activation of astrocytes derived from rats, which may be involved in the block-ade of STAT3 phosphorylation.关键词
星形胶质细胞/脑出血/表皮生长因子受体/小干扰RNA/神经胶质原纤维酸性蛋白/信号传导蛋白和转录激活物3Key words
astrocytes/intracerebral hemorrhage/epi-dermal growth factor receptor/siRNA/glial fibrillary acidic protein/signal transducers and activators of transcription 3分类
医药卫生引用本文复制引用
钱红,刘理静,李艳春,谢明,武衡,刘双喜,武斌..EGFR siRNA通过阻碍STAT3磷酸化抑制大鼠星形胶质细胞活化[J].中国药理学通报,2016,(2):216-222,7.基金项目
湖南省教育厅科研项目(No 14C0906) （No 14C0906）
