EGFR siRNA inhibits activation of astrocytes derived from rats through blockade of STAT3 phosphorylation
10.3969/j.issn.1001-1978.2016.02.014
- VernacularTitle:EGFR siRNA通过阻碍STAT3磷酸化抑制大鼠星形胶质细胞活化
- Author:
Hong QIAN
;
Lijing LIU
;
Yanchun LI
;
Ming XIE
;
Heng WU
;
Shuangxi LIU
;
Bin WU
- Publication Type:Journal Article
- Keywords:
astrocytes;
intracerebral hemorrhage;
epi-dermal growth factor receptor;
siRNA;
glial fibrillary acidic protein;
signal transducers and activators of transcription 3
- From:
Chinese Pharmacological Bulletin
2016;(2):216-222
- CountryChina
- Language:Chinese
-
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.
