CLEC-2在中枢神经系统炎症中的表达意义

The expression significance of CLEC-2 in inflammation of CNS

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.