整合素β8在星形胶质细胞氧糖剥夺后对TGF-β1的活化影响  

Effect of integrin β_8 on TGF-β1 activation in astrocytes with oxygen glucose deprivation

作　　者：李晋辉[1] 李德渊[1] 陈大鹏[1] 母得志[1] 屈艺[1] 

机构地区：[1]四川大学华西第二医院儿科/华西儿童医学中心,出生缺陷相关妇儿疾病教育部重点实验室,四川成都610041

出　　处：《中国当代儿科杂志》2014年第1期73-76,共4页Chinese Journal of Contemporary Pediatrics

基　　金：国家重大科学研究计划项目(2013CB967404);国家自然科学基金(31171020、81172174、81270724、81100457);四川省科技厅科技支撑项目(2010SZ0280、2013SZ0031);国家临床重点专科(儿科新生儿专业)建设项目

摘　　要：目的探讨在氧糖剥夺(OGD)后,星形胶质细胞中整合素β8(β8)的表达对转化生长因子-β1(TGF-β1)活化的影响。方法体外培养星形胶质细胞,OGD模拟缺氧缺血。免疫细胞化学法检测β8蛋白在星形胶质细胞中的表达及分布,Western blot定量检测OGD处理后复氧12 h、1 d及2 d时β8蛋白表达的变化。建立星形胶质细胞与荧光素酶报道细胞(TMLC)共培养体系,利用RNA干扰技术抑制星形胶质细胞β8表达,观察β8对TGF-β1活化的影响。结果β8主要分布于星形胶质细胞的胞浆和树突;星形胶质细胞中β8蛋白表达在复氧后12 h即有增加,1 d时达高峰;星形胶质细胞与TMLC共培养体系在复氧后各时间点TGF-β1活性增高趋势与星形胶质细胞β8表达趋势相似,抑制β8表达后,TGF-β1活性在各时间点均显著降低。结论新生鼠在缺血缺氧时,星形胶质细胞高表达的β8对中枢神经系统中TGF-β1的活化起重要调控作用。Objective To study the effect of β8 expression on transforming growth factor β1（TGF-β1） activation in astrocytes with oxygen glucose deprivation （OGD）. Methods Astrocytes were cultured and then subjected to OGD to generate hypoxia-ischemia （HI） model in vitro. Immunocytochemistry was used to detect the expression and distribution of β8 in nomoxia cultured cells. β8 protein expression was quantified by Western blot at 12 hours, 1 day and 2 days after OGD. Astrocytes and luciferase reporter cells （TMLC） were co-cultured. β8 RNA interference system was established to specifically inhibit β8 expression in cultured astrocytes. TGF-β1 activation was then detected in the co- culture system. Results β8 was mainly located in the cytoplasm and neurites of astrocytes. OGD resulted in increase of β8 protein expression at 12 hours after reoxygenation in astrocytes, which was peaked at 1 day after reoxygenation. TGF-β1 activation was in accordance with β8 expression in astrocyte-TMLC co-culture system after reoxygenation. After the inhibition of β8, TGF-β1 activation was significantly reduced in all time points. Conclusions The highly expressed β8 plays important roles in the regulation of TGF-β1 activation in neonatal rats with hypoxic-ischemic brain damage.

关 键 词：缺氧缺血 星形胶质细胞 整合素 转化生长因子 新生大鼠 

分 类 号：R722.1[医药卫生—儿科]