摘要
目的探讨siRNA沉默表皮生长因子受体(EGFR)表达对大鼠脑出血后脑损伤的影响及相关机制。方法以Ⅶ型胶原酶注入苍白球构建大鼠脑出血模型,予10μL空病毒载体或EGFR siRNA慢病毒表达载体侧脑室注射,进行神经功能评分,应用HE染色观察脑组织病理改变,干湿重法测定脑组织含水量,免疫组织化学染色测定脑组织中EGFR表达,荧光实时定量PCR分析神经胶质原纤维酸性蛋白(GFAP)mRNA水平,Western blot检测EGFR、GFAP、信号传导蛋白和转录激活物3(STAT3)、磷酸化STAT3(p-STAT3)蛋白表达。结果随着脑出血时间的延长,血肿周围脑组织EGFR表达逐渐上调,第7天达到高峰(P<0.01)。与模型组比较,EGFR siRNA缓解脑组织病理损害,减少神经功能评分、脑组织含水量,并降低血肿周围脑组织EGFR、GFAP及p-STAT3表达水平(P<0.01)。结论 EGFR基因沉默通过阻碍STAT3磷酸化抑制星形胶质细胞活化,进而减轻大鼠脑出血后脑损伤。
Objective To explore the effects of siRNA for epidermal growth factor receptor（ EGFR） on rat brain injury after intracerebral hemorrhage and its related mechanisms. Methods The intracerebral hemorrhage model was established by injecting of Ⅶ collagenase into the caudate nucleus in rats. Rats were then treated with injection of 10 μL empty viral vector or EGFR siRNA lentivirus expression vector into lateral ventricles. The neurological function score was evaluated,and HE staining was used to observe the pathological changes of cerebral tissues.Water content in brain was examined by the dried and wet weight method. EGFR expression was measured using immunohistostaining. Glial fibrillary acidic protein（ GFAP） mRNA level was analyzed through fluorescence realtime quantitative PCR. Western blot was used to detect EGFR,GFAP,signal transducers and activators of transcription 3（ STAT3） and phosphorylated STAT3（ p-STAT3）. Results With the extension of intracerebral hemorrhage time,ERGF expression in cerebral tissues around hemotomas was gradually upregulated,and reached thepeak on day 7（ P〈 0. 01）. In comparison with model group,EGFR siRNA relieved pathological impairment of cerebral tissues,reduced neurological function score and brain water content,and decreased the expression levels of EGFR,GFAP and p-STAT3 in cerebral tissues around hemotomas（ P〈 0. 01）. Conclusions Gene silence of EGFR inhibits the activation of astrocytes through blockade of STAT3 phosphorylation,thus leading to reduction of brain injury after intracerebral hemorrhage.
出处
《基础医学与临床》
CSCD
2016年第12期1681-1686,共6页
Basic and Clinical Medicine
基金
湖南省教育厅资助科研项目(14C0906)
