目的:研究C型凝集素家族重要成员C型凝集素样受体2(C-type lectin-like receptor-2,CLEC-2)在中枢神经系统(central nervous system,CNS)炎症反应中的表达变化及其生物学作用。方法:在大鼠侧脑室内注射脂多糖(lipopolysaccharide,LPS)构建大脑炎症模型,运用Western Blot方法检测CLEC-2在LPS侧脑室内注射的大脑炎症及LPS诱导的原代小胶质细胞炎性活化模型中的蛋白表达变化;运用免疫荧光方法检测CLEC-2在大脑皮层中的细胞定位。结果:Western Blot显示大脑皮层中CLEC-2的蛋白表达呈现先上升后下降的趋势。免疫荧光分析显示CLEC-2与小胶质细胞存在共定位,且侧脑室注射LPS 12 h后CLEC-2在小胶质细胞中的表达增加。在体外原代小胶质细胞活化模型中,CLEC-2和P-Syk蛋白的表达水平也明显升高。结论:脑炎症中CLEC-2表达上调,其主要定位在活化的小胶质细胞上,表明在中枢神经系统炎症中CLEC-2可能发挥着重要的作用。
Objective: To investigate the expression and localization changes of C-type lectin domain family 2(CLEC-2) in lipopolysaccharide(LPS) intracerebroventricular injected cerebral cortexs to demostrate the role of CLEC-2 in the regulation of central nervous system(CNS) inflammation. Methods: The model of brain and primary rat cerebral cortex microglia inflammation was prepared in rats by the stimulation of LPS. The content of CLEC-2 protein were assayed by Western Blot. The location of CLEC-2 in cerebral cortexs were detected by double immumofluorescence. Results: Following LPS intracerebroventricular injection, expression of CLEC-2 in the cerebral cortexs of LPS intracerebroventricular injected rats was upregulated as compared to the nomal cerebral cortexs. Double labeling demonstrated the elevated CLEC-2 mostly located in active microglia. The expression of CLEC-2 was increased in the cerebral cortexs microglia after stimulated with LPS. Furthermore, the expression of CLEC-2 in primary rat cerebral cortex microglia was increased after LPS stimulation, as well as the expression of P-Syk. Conclusions: Injection of LPS within intracerebroventricular upregulated CLEC-2 expression in the whole inflammatory phase. In the cerebral cortex of rats, CLEC-2 immunoreactivity was mostly co-localized with activated microglia, indicating that CLEC-2 may play an important role in the inflammation of CNS.