沙眼衣原体持续和急性感染状态McCoy细胞Rab蛋白表达差异的研究  

Differences in Rab protein expressions in McCoy cells with acute versus persistent Cldamydia trachomatis infection

作　　者：马春光[1] 朱慧玲[2] 张馨月[1] 陈木开[1] 韩建德[1] 

机构地区：[1]中山大学附属第一医院皮肤科,广州510080 [2]广州医科大学附属第一医院皮肤科

出　　处：《中华皮肤科杂志》2016年第5期334-337,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81071404）;广东省自然科学基金（S2013010015360）;广州市科技计划项目（2014J4100178）

摘　　要：目的探讨沙眼衣原体（Ct）急性感染和持续感染McCoy细胞后Rab蛋白家族表达的差异。方法ct感染McCoy细胞后，经青霉素G诱导获得持续感染组，未经青霉素G诱导为急性感染组，同时设立无衣原体感染、无青霉素G处理的空白对照组和无衣原体感染、有青霉素G处理的青霉素对照组。感染48h后，裂解McCoy细胞、收集蛋白后，用ELISA检测以上各组Rab4A、Rab6A、Rab10、Rab11A、Rab14蛋白水平，同时提取各组细胞RNA，采用荧光定量PCR检测Rab4A和Rabl4mRNA的相对表达量（用2。坳法计算）。结果ct持续感染组Rab4A、Rab6A、Rab10、Rab11A、Rab14蛋白水平均高于急性感染组，差异有统计学意义（z值均为3．621，均P〈0．001），且急性感染组和持续感染组中5种蛋白水平均显著低于空白对照组（均P〈0．0083），而空白对照组和青霉素对照组中这5种Rab蛋白表达水平差异无统计学意义（均P〉0．05）。在转录水平，Rab4A和Rab14mRNA在各组的表达均显示与其蛋白表达相同的趋势。结论McCoy细胞在ct持续感染状态下，Rab蛋白的转录和表达水平高于急性感染状态。Objective To investigate differences in Rab protein expressions in McCoy cells with acute versus persistent Chlamydia trachomatis （Ct） infection. Methods Cultured McCoy cells were infected with different amounts （400, 500, 550 μl/well） of Ct strain D suspensions , then cultured with the medium containing 100 U/ml penicillin G （persistent Ct infection groups） or that without penicillin G （acute Ct infection groups）. Ct-uninfected McCoy cells receiving no penicillin G treatment served as the blank control group, and those receiving penicillin G treatment as the penicillin group. After 48-hour culture, McCoy cells were lysed, proteins were collected, and total RNA was extracted from the cells. Enzyme-linked immunosorbent assay （ELISA） was conducted to measure protein levels of Rab4A, Rab6A, Rabl0, Rab11A and Rab14, and fluorescence-based quantitative PCR to quantify mRNA expressions of Rab4A and Rab14 （expressed as 2-△△O）. Results Protein levels of Rab4A, Rab6A, Rabl0, Rabl 1A and Rab14 were all significantly lower in the acute than in the persistent Ct infection groups （all Z = 3.621, P 〈 0.001 ）, and lower in the persistent and acute Ct infection groups than in the blank control group （all P 〈 0.008 3 ）, but insignificantly different between the blank control group and penicillin group （all P 〉 0.05）. In addition, the expressions of Rab4A and Rab14 mRNAs were consistent with those of their proteins in these groups. Conclusion The transcriptional and expression levels of Rab proteins are higher in McCoy cells persistently infected with Ct than in those acutely infected with Ct.

关 键 词：沙眼衣原体 衣原体感染 RAB GTP结合蛋白质类 急性感染 持续感染 McCoy细胞 

分 类 号：R518[医药卫生—内科学]