Differential Expression of mRNA Encoding Cytokines and Chemokines in the Reproductive Tract after Infection of Mice with Chlamydia trachomatis

K. L. Cerny, Maranda Van Fleet, Anatoly V. Slepenkin, E. Peterson, P. Bridges
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引用次数: 5

Abstract

Infection with Chlamydia trachomatis targets epithelial cells within the genital tract which respond by secreting chemokines and cytokines. Persistent inflammation can lead to fibrosis, tubal infertility and/or ectopic pregnancy; many infections are asymptomatic. Most studies have investigated the inflammatory response in the initial stages of infection, less is known about the later stages of infection, especially with a low, potentially asymptomatic, bacterial load. Our objective was to determine the inflammatory mediators involved in clearance of low-grade infection and the potential involvement in chronic inflammation. Six to eight week old C3H/HeJ mice were pretreated with 2.5 mg medroxyprogesterone acetate on day -10 and -3 before infection. Mice (n=3 for 28 d, n=3 for 35 d) were infected with 5 × 102 inclusion-forming units of C. trachomatis, serovar D; vaginal cultures were obtained weekly to monitor infection. Control mice (n=3 for 28 d, n=3 for 35 d) were sham infected. Mice were killed on day 28 (experiment 1) and day 35 (experiment 2) post-infection and vaginal tissue, uterine horns and oviducts collected for analysis of mRNAs encoding inflammatory cytokines and chemokines. Total RNA was isolated and a superarray analysis performed using mouse Cytokines and Chemokines PCR arrays (Qiagen, Valencia, CA). Statistical differences in gene expression were determined using a paired Students t-test. At 28 days after infection, the expression of mRNA encoding 6, 35 and 3 inflammatory genes differed from controls in vaginal, uterine and oviductal tissues, respectively (P<0.05). At 35 days after infection, the expression of mRNA encoding 16, 38 and 14 inflammatory genes differed from controls in vaginal, uterine and oviductal tissues, respectively (P<0.05). Understanding the mechanisms involved in the inflammatory response at later stages of infection should aid in the development of treatment options that minimize the development of asymptomatic, chronic inflammation-induced infertility.
沙眼衣原体感染小鼠生殖道细胞因子和趋化因子mRNA的差异表达
沙眼衣原体感染的目标是生殖道内的上皮细胞,其反应是分泌趋化因子和细胞因子。持续的炎症可导致纤维化、输卵管不孕和/或异位妊娠;许多感染是无症状的。大多数研究都研究了感染初期的炎症反应,但对感染后期,特别是低水平、可能无症状的细菌载量,了解较少。我们的目的是确定参与清除低级别感染的炎症介质和慢性炎症的潜在参与。6 ~ 8周龄C3H/HeJ小鼠在感染前第10天和第3天分别给予2.5 mg醋酸甲孕酮预处理。小鼠(n=3, 28 d, n=3, 35 d)感染5 × 102个包涵体单位沙眼衣原体,血清型d;每周进行阴道培养以监测感染情况。对照小鼠(n=3只,28 d, n=3只,35 d)进行假感染。在感染后第28天(实验1)和第35天(实验2)处死小鼠,收集阴道组织、子宫角和输卵管,分析编码炎症细胞因子和趋化因子的mrna。分离总RNA,并使用小鼠细胞因子和趋化因子PCR阵列(Qiagen, Valencia, CA)进行超阵列分析。使用配对学生t检验确定基因表达的统计差异。感染后28 d,阴道、子宫和输卵管组织中编码6、35和3个炎症基因的mRNA表达量分别高于对照组(P<0.05)。感染后35 d,阴道、子宫和输卵管组织中编码16、38和14个炎症基因的mRNA表达量分别高于对照组(P<0.05)。了解感染后期炎症反应的机制应该有助于制定治疗方案,最大限度地减少无症状慢性炎症性不孕的发展。
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