中华皮肤科杂志 ›› 2019, Vol. 52 ›› Issue (8): 554-560.doi: 10.3760/cma.j.issn.0412-4030.2019.08.008

• 论著 • 上一篇    下一篇

不同途径接种衣原体后小鼠多脏器衣原体检测分析

邵丽丽    马璟玥    练婷婷    魏世娟    任杰    刘全忠   

  1. 天津医科大学总医院皮肤科  天津性传播疾病研究所  300052
  • 收稿日期:2018-08-16 修回日期:2019-03-15 出版日期:2019-08-15 发布日期:2019-07-30
  • 通讯作者: 刘全忠 E-mail:liuqztjykdxzyy@163.com
  • 基金资助:
    国家自然科学基金(31500157)

Detection of Chlamydia in multiple visceral organs of mice inoculated with Chlamydia through different routes

Shao Lili, Ma Jingyue, Lian Tingting, Wei Shijuan, Ren Jie, Liu Quanzhong   

  1. Department of Dermatology, General Hospital of Tianjin Medical University, Institute of Sexually Transmitted Diseases, Tianjin 300052, China
  • Received:2018-08-16 Revised:2019-03-15 Online:2019-08-15 Published:2019-07-30
  • Contact: Liu Quanzhong E-mail:liuqztjykdxzyy@163.com
  • Supported by:
    National Natural Science Foundation of China (31500157)

摘要: 【摘要】 目的 验证衣原体能够从小鼠生殖道传播至胃肠道,并在胃肠道长期定植。方法 5 ~ 6周龄雌性C57BL/6J小鼠120只,分为阴道接种组(35只)、灌胃接种组(30只)、肛门直肠接种组(30只)、眶后静脉丛接种组(5只),感染鼠衣原体,同时每组均设阴性对照,5只/组。各组小鼠均在感染后第3、7天及此后每隔7天用拭子取阴道及直肠分泌物,检测拭子中脱落细胞感染衣原体的数量。应用间接免疫荧光、定量PCR检测接种后第7、14、28、56、105天阴道接种组小鼠生殖道(阴道、子宫、输卵管及卵巢)、胃肠道(胃、小肠、盲肠、结肠、直肠)、肠外组织(心、肝、脾、肺、肾)中的衣原体活菌量及基因组(活菌量与基因拷贝数以10为底数对数转换),观察生殖道输卵管积水及炎症程度、胃肠道的组织病理变化;检测接种后第28天、56天灌胃接种组、肛门直肠接种组小鼠衣原体在生殖道及胃肠道的感染及致病情况。眶后静脉丛接种组在接种后的第3、5、7、10、14天小鼠尾静脉取血,检测血中衣原体的活菌量及基因组,观察感染第56天时生殖道、胃肠道致病情况。结果 阴道接种后第7天,所有小鼠的生殖道、胃肠道及肠外组织均检测到活菌和基因组,阴道的菌量及其基因拷贝数常用对数值最高分别为6.26 ± 0.56、7.30 ± 0.23,直肠的菌量及其基因拷贝数常用对数值分别为2.60 ± 1.95、4.87 ± 0.09;第28天时,心脏、肺等肠外组织中均未检测到活菌,生殖道和胃肠道组织仍可检测到活衣原体,阴道菌量及其基因拷贝数常用对数值分别为3.47 ± 1.06、5.80 ± 1.49,直肠菌量及其基因拷贝数常用对数值分别为4.00 ± 0.35、5.14 ± 0.81;第56天时,仅在胃肠道组织中检测到活病原体;第105天,在胃肠道中仍检测到衣原体活菌和基因组,直肠的菌量及其基因拷贝数常用对数值可分别达到2.60 ± 0.65、4.29 ± 0.57。灌胃接种组、肛门直肠接种组小鼠,在接种第28、56天时,所有小鼠胃肠道均检测到衣原体活菌和基因组。眶后静脉丛接种组衣原体在血液中生存14 d左右,第14天所有小鼠均在肛门直肠拭子中检测到活菌。阴道接种鼠衣原体56 d后,5只小鼠的生殖道形成严重的输卵管积水及慢性炎症,输卵管扩张,但胃肠道未见到明显的炎症细胞浸润;其他接种途径亦未在胃肠道观察到炎症病理变化。结论 衣原体生殖道感染后可致系统播散,能够传播至胃肠道,在胃肠道定植并长期生存。

关键词: 衣原体感染, 沙眼衣原体, 鼠衣原体, 胃肠道, 生殖道感染, 定植

Abstract: 【Abstract】 Objective To certify that Chlamydia can spread from the genital tract to the gastrointestinal tract for long-lasting colonization. Methods Totally, 120 female C57BL/6J mice aged 5 - 6 weeks were divided into 4 experimental groups to be inoculated with purified Chlamydia muridarum (C. muridarum) elementary bodies in the vagina (n = 35), gastric area (n = 30), anus and rectum (n = 30), retro-orbital venous plexus (n = 5) respectively. Moreover, corresponding negative groups inoculated with sucrose phosphate glutamate buffer (n = 5) were set up for each experimental group. On days 3, 7, and every 7 days, vaginal and rectal discharges were collected with swabs from the mice, and the number of live C. muridarum orgnisms in exfoliated cells infected with C. muridarum in the swabs was determined. Indirect immunofluorescence assay and quantitative PCR (qPCR) were performed to determine the number of live chlamydial organisms and the copy number of chlamydial genomes in the mouse genital tract (vagina, uterus, oviduct and ovary), gastrointestinal tract (stomach, small intestine, cecum, colon, rectum) and parenteral tissues (heart, liver, spleen, lung, kidney) on days 7, 14, 28, 56 and 105 after the inoculation. The number of live chlamydial organisms and copy number of chlamydial genomes were transformed logarithmically with a base of 10. The degree of hydrosalpinx and inflammation in the genital tract, and histopathological changes of the gastrointestinal tract were observed. The infectivity and virulence of C. muridarum in the genital tract and gastrointestinal tract were evaluated in the intragastric inoculation group and intra-anal and intrarectal inoculation group on days 28 and 56 after the inoculation. Blood samples were obtained from the mouse caudal vein in the retro-orbital venous plexus inoculation group on days 3, 5, 7, 10 and 14 after the inoculation, the number of live chlamydial organisms and the copy number of chlamydial genomes in the blood samples were determined, and chlamydial infectivity in the genital tract and gastrointestinal tract was evaluated on day 56. Results On day 7 after the inoculation in the vagina, both C. muridarum live organisms and genomes were detected in the genital tract, gastrointestinal tract and parenteral tissues of all the mice. The largest common logarithm of the number of C. muridarum inclusion forming units (IFU) was observed in the vagina (6.26 ± 0.56), with the common logarithm of the copy number of chlamydial genomes in the vagina being 7.30 ± 0.23, and the common logarithms of the number of Chlamydia IFU and genomic copy were 2.60 ± 1.95 and 4.87 ± 0.09 respectively in the rectum. On day 28, no live Chlamydia was detected in the heart, lung or other parenteral tissues, while live Chlamydia could be found in the genital tract and gastrointestinal tract. The common logarithms of the number of Chlamydia IFU and genomic copy were 3.47 ± 1.06 and 5.80 ± 1.49 respectively in the vagina, and 4.00 ± 0.35 and 5.14 ± 0.81 respectively in the rectum. On day 56, live Chlamydia could only be detected in the gastrointestinal tract. On day 105, live Chlamydia and its genomes could be still detected in the gastrointestinal tract, and the common logarithms of the number of Chlamydia IFU and genomic copy could be up to 2.60 ± 0.65 and 4.29 ± 0.57 respectively in the rectum. On days 28 and 56 after the inoculation, both live Chlamydia and its genomes could be detected in the gastrointestinal tract of all the mice in the intragastric inoculation group and intra-anal and intrarectal inoculation group. Chlamydia could survive in the blood for about 14 days in the retro-orbital venous plexus inoculation group, and live Chlamydia was detected in anal-rectal swabs in all the mice on day 14. On day 56 after the intravaginal inoculation with C. muridarum, severe hydrosalpinx, chronic inflammation and oviduct dilation occurred in the genital tract of 5 mice, but there was no obvious infiltration of inflammatory cells in the gastrointestinal tract, and inflammatory pathological changes were not observed in the gastrointestinal tract of mice after inoculation with Chlamydia through other routes either. Conclusion The infection with Chlamydia in the genital tract can lead to systemic dissemination, and Chlamydia can be spread to the gastrointestinal tract, and colonize and survive in the gastrointestinal tract for a long time.

Key words: Chlamydia infections, Chlamydia trachomatis, Chlamydia muridarum, Gastrointestinal tract, Reproductive tract infections, Colonization