COP9信号小体基因缺失对新型隐球酵母菌嗜中枢性的影响与机制研究

Influence of COP9 signalosome complex gene deletion on addicted to infringe central nervous system of Cryptococcus neoformans and its mechanism

目的探讨COP9信号小体（CSN）基因缺失对新生隐球酵母菌嗜中枢性的影响及可能机制,为研究抗真菌药物作用靶点提供实验数据。方法 2014年7月以环磷酰胺免疫抑制C57BL/6小鼠,尾静脉注射H99株、COP9信号小体CSN1201基因缺失株及生理盐水,构建小鼠静脉注射感染模型H99组、CSN1201△及对照组各10只;所有数据采用SPSS 15.0进行分析。结果 H99组小鼠脑组织真菌培养在12h即出现菌落生长,生长速度迅速,PAS染色可见隐球酵母菌菌体;CSN1201△组及对照组脑组织培养结果为阴性,PAS染色未见隐球酵母菌菌体;H99组小鼠脑组织RhoA蛋白表达水平值随时间增加而增加,与对照组相比,24~72h该蛋白表达显著增加（P〈0.05）;Rac1及Cdc42蛋白表达未随时间变化,但与对照组相比蛋白表达也明显增加（P〈0.05）。结论COP9信号小体CSN1201基因缺失能够降低新生隐球酵母菌穿越血脑屏障的能力,其影响途径可能与脑微血管内皮细胞紧密连接蛋白细胞骨架重排的调控有关。

OBJECTIVE To explore the influence of COP9signalosome（CSN）complex gene deletion on addicted to infringe central nervous system of Cryptococcus neoformans and analyze the possible mechanisms so as to provide experimental data for research on the targets of antifungal drugs.METHODS The cyclophosphamide immunosuppressive C57BL/6mice were treated with tail intravenous injection of H99 strains,COP9signalosome CSN1201 gene deletion strains,and normal saline so as to build the mice intravenous injection infection models：the H99 group,the CSN1201 △ group,and the control group,with 10 mice in each.All of the data were statistically analyzed by using SPSS 15.0software.RESULTS The colony growth of the H99 group was found after the fungal culture of mice brain tissues was performed for 12 hours,with the growth rate rapid,and C.neoformans strains were displayed by PAS staining.The culture of brain tissues was negative in the CSN1201△ group and the control group,and C.neoformans strains were not displayed by PAS staining.The level of expression of RhoA protein in the mice brain tissues of the H99 group was increased with the time,as compared with the control group,it was increased significantly after the culture for 24-72 hours.The level of expression of Rac1 or Cdc42protein did not change with the time,however,as compared with the control group,they were increased significantly（P0.05）.CONCLUSIONThe COP9 signalosome CSN1201gene deletion may reduce the C.neoformans ability to pass through the blood-brain barrier,and the effect pathway may be associated with the regulation of brain microvascular endothelial cells tight junction protein cytoskeleton rearrangement.