目的:观察深部致病性真菌新生隐球菌分泌的胞外蛋白水解酶对脑微血管内皮细胞中基质金属蛋白酶(MMP)-9、微管相关蛋白(Tau—LRP)表达的影响以及抑肽酶的干预作用。方法:在含有成熟的脑微血管内皮细胞的培养皿中,分别加入胞外蛋白水解酶相关成分及其特异性抑制剂后,应用免疫组化技术检测内皮细胞中MMP-9、Tau—LRP表达的变化。结果:加入新生隐球菌分泌的胞外蛋白水解酶和(或)丝氨酸蛋白酶后内皮细胞的MMP-9、Tau—LRP表达均升高,与对照组比较,差异有统计学意义(P〈0.01);抑制剂组的MMP-9、Tau—LRP与对照组比较,差异无统计学意义(P〉0.05)。结论:新生隐球菌分泌的胞外蛋白水解酶通过上调MMP-9和(或)Tau—LRP表达,诱导微血管内皮细胞基质降解和细胞自身微管结构及紧密连接发生变化,可能是最终导致血脑屏障通透性增加的原因之一。
Objective: To study the influence of extracellular proteinase in the supernatant in the mediums for culturing deep pathogenic fungus Cneoformans on the expression of matrix metalloproteinase-9 (MMP-9) and Tau-LRP of brain microvascular endothelial cells, and the effect of interference of aprotinin. Methods: Adding the supernatant with or without aprotinin into cultured plate containing mature brain microvascular endothelial cells of rat respectively. The expression of MMP-9 and Tau-LRP was measured by immunohistochemical assay. Results: After adding the supernatant, the expression of MMP-9 and Tau-LRP was markedly increased compared with that in the control group (P〈 0.01). In aprotinin-adding group, the MMP-9 and Tau-LRP expression had no significant difference compared with that in the control group (P〉 0.05). Conclusion: Extracellular proteinase in the supernatant might up-regulates the MMP-9 and Tau-LRP expression of microvascular endothelial cells, which may cause increased permeability of brain-blood barrier.