新生隐球菌通过S100A10活化尿激酶-纤溶酶系统侵袭血脑屏障的机制研究

中国真菌学杂志　 2016, Vol. 11 　Issue (3): 129-134.

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新生隐球菌通过S100A10活化尿激酶-纤溶酶系统侵袭血脑屏障的机制研究

邱天文, 朱红梅, 温海

上海市长征医院皮肤科上海市医学真菌分子生物学重点实验室, 上海 200003

收稿日期:2016-05-16 出版日期:2016-06-28 发布日期:2016-06-28
通讯作者: 温海,E-mail:wenhaipfk@126.com;朱红梅,E-mail:hmzhu_cn@163.com E-mail:wenhaipfk@126.com;hmzhu_cn@163.com
作者简介:邱天文,男(汉族),硕士研究生在读.E-mail:872280290@qq.com
基金资助:
国家自然科学基金（31470252，81271800，31270181）

Cryptococcus traversal across the Blood-Brain Barrier though S100A10-dependent activation of urokinase-plasmin system

QIU Tian wen, ZHU Hong-mei, WEN Hai

PLA Key Laboratory of Fungal Diseases and Shanghai Key Laboratory of Molecular Medical Mycology, Department of Dermatology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China

Received:2016-05-16 Online:2016-06-28 Published:2016-06-28

摘要/Abstract

摘要：

目的验证隐球菌可以上调脑微血管内皮细胞S100A10基因来活化尿激酶-纤溶酶系统，从而更易穿过血脑屏障。方法 ①将小鼠脑微血管内皮细胞bEnd.3和新生隐球菌B3501共孵育，检测加菌的实验组和不加菌对照组S100A10、UPA基因和蛋白的表达量。②用慢病毒感染血管内皮细胞，比较S100A10基因正常组和沉默组的S100A10和UPA基因和蛋白的表达量。③构建Transwell血脑屏障模型，比较S100A10基因正常组和沉默组隐球菌通过的差异性，用底物显色法检测尿激酶-纤溶酶系统的相关活化情况。结果 ①新生隐球菌B3501和bEnd.3共孵育，Real-time PCR显示S100A10和UPA的mRNA高表达；Western-blot显示S100A10目的蛋白和UPA蛋白高表达。②沉默S100A10基因后，S100A10和UPA基因及蛋白表达分别受到抑制。③隐球菌添加到Transwell体外血脑屏障模型中，S100A10基因未沉默组通过血脑屏障的隐球菌数量比S100A10基因沉默组多，尤其是8 h、16 h、24 h，两组差异有统计学意义，P<0.05；未沉默S100A10基因的对照组的纤溶酶原和尿激酶的活化程度高于沉默S100A10基因的实验组，其中纤溶酶原在8 h、16 h、24 h，尿激酶在16 h、24 h，两组差异有统计学意义，P<0.05。结论隐球菌能同时使内皮细胞S100A10基因、UPA基因的高表达，活化尿激酶-纤溶酶系统，提高其通过血脑屏障的能力。

关键词: 隐球菌, S100A10, 尿激酶, 纤溶酶, 血脑屏障

Abstract:

Objective To find if Cryptococcus traversal across the blood-brain barrier though S100A10-dependent activation of urokinase-plasmin system.Methods ①The mouse brain microvascular endothelial cells bEnd.3 and Cryptococcus neoformans B3501 were co-incubated.Then the expression of S100A10 gene and UPA genewere tested.②Expressionof S100A10 and UPA in S100A10 normal and silent groups of endothelial cellsInfected with lentivirus were also compared.③The abilities of Cryptococcus S100A10 normal and silent groups traversal across the transwell blood-brain barrier model were compared.Urokinase-related activation of plasminogen was tested by the chromogenic substrate assay.Results ①In bEnd.3 incubated with Cryptococcus neoformans B3501,Real-time PCR display high expression of mRNA of S100A10 and UPA;Western-blot show S100A10 protein and UPA protein overexpression.②While S100A10 gene been silenced,S100A10 and UPA expressions were suppressed.③Cryptococcal traversal across the transwell in vitro BBB model in S100A10 silenced group was less than that of controlgroup.The diffences on 8 h,16 h,24 h were especially statistically significant,P<0.05.The activation of urokinase and plasmin was also lower than that of control group.The diffences of plasminogen activation on 8 h,16 h and 24 h in two groups,the diffences of urokinase activation on 16 h and 24 h in two groups,were statistically significant,respectively.Conclusion Cryptococcus traversal across the blood-brain barrier might be though S100A10-dependent activation of urokinase-plasmin system.In the meantime,the S100A10 gene and UPA gene of brain microvascular endothelial cells were highly expressed.

Key words: Cryptococcus neoformans, S100A10, urokinase, plasmin, blood-brain barrier

中图分类号:

R379.5

邱天文, 朱红梅, 温海. 新生隐球菌通过S100A10活化尿激酶-纤溶酶系统侵袭血脑屏障的机制研究[J]. 中国真菌学杂志, 2016, 11(3): 129-134.

QIU Tian wen, ZHU Hong-mei, WEN Hai. Cryptococcus traversal across the Blood-Brain Barrier though S100A10-dependent activation of urokinase-plasmin system[J]. Chinese Journal of Mycology, 2016, 11(3): 129-134.

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新生隐球菌通过S100A10活化尿激酶-纤溶酶系统侵袭血脑屏障的机制研究

Cryptococcus traversal across the Blood-Brain Barrier though S100A10-dependent activation of urokinase-plasmin system

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