Disorder of the Microbiota and Mucosal Immunity of the Genital Tract in Women of Reproductive Age with Different Clinical Courses of Acute Uncomplicated Pyelonephritis

Q4 Biochemistry, Genetics and Molecular Biology
A.V. Rudenko, A.M. Romanenko, S.P. Pasiechnikov, M.V. Mitchenko, O.V. Romashchenko, N.M. Taradiy
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Methods. 246 women of reproductive age suffering from acute uncomplicated pyelonephritis were studied, divided into three variants of the disease clinical course: group 1 - mild (n=105), group 2 - moderate (n=90), and group 3 - severe (n=51) courses. The scrapings of the mucous membrane of the cervical canal and washings from the vagina were obtained before the start of antimicrobial therapy. Quantitative determination of pathogens was carried out by sowing the biological material on solid nutrient environment. Identification of detached bacteria was carried out according to Bergeys. Mollicutes (Ureaplasma spp., Mycoplasma hominis) were determined by the cultural-fermentative method using test systems and the polymerase chain reaction. Vaginal washings of 121 sick women were used for immunological studies. The levels of myeloperoxidase, lysozyme, human β-defensin-2, immunoglobulins M, A, G, secretory IgA, lactoferrin, C3-component of complement, secretory component, and tumor necrosis factor-α were determined. The reference group consisted of 23 clinically healthy women. The Statistica 12.0 program package for Windows was used, and a difference of p<0.05 was considered verifi able. Results. In the majority of patients with acute uncomplicated pyelonephritis (85.6% of cases), concomitant chronic pelvic inflammatory diseases (colpitis, salpingitis, underlying medical condition of cervix) were observed. Classical bacteria were more often detected in vaginal washings of patients of all groups, while mollicutes were detected in scrapings of the mucous membrane of the cervical canal, whereas the frequency of their detection increased with increase in the pyelonephritis severity. The analysis of indicators of local immunity determined disorders of mucosal immunity of the genital tract in the form of increased levels of myeloperoxidase, IgA, and IgG relative to reference values. The highest levels of lysozyme and myeloperoxidase were determined in patients with a severe course of acute uncomplicated pyelonephritis (by 5.3 and 3.6 times more, respectively), and the content of immunoglobulins A, M, and G in patients with a mild course of the disease (by 3.2, 3.1, and 4.0 times more, respectively). An increase in the level of tumor necrosis factor-α was detected in all patients, although no significant differences from controls were found in any group, but the highest median value was recorded in patients with severe pyelonephritis. In the group of patients with a severe clinical course, the lowest median was observed for β-defensin-2, which makes it impossible to inhibit the synthesis of TNF-α and, thus, supports the inflammatory process. Conclusions. It has been proven that in women with acute uncomplicated pyelonephritis, the severity of the course probably correlates with infection of the genital tract by mollicutes, mainly in association with classical bacteria and with level of mucosal immunity disorder. The obtained results of microbiological and immunological studies of biological material collected before the start of antibacterial therapy in patients with acute uncomplicated pyelonephritis proved the need for a mandatory examination by a gynecologist to ascertain concomitant chronic pelvic inflammatory diseases and establish infection with mollicutes to provide relevant etiotropic treatment. 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引用次数: 0

Abstract

The research deals with the study of immuno-microbiological parallels of the genital tract in women with acute uncomplicated pyelonephritis with concomitant chronic pelvic inflammatory diseases. In addition to etiotropic antibacterial therapy, correction of the mucosal immunity state of the genital tract is crucial for the treatment of the disease and prevention of its recurrence. The purpose of the work was to investigate the contamination and state of local immunity of the genital tract in women with acute uncomplicated pyelonephritis with different clinical courses. Methods. 246 women of reproductive age suffering from acute uncomplicated pyelonephritis were studied, divided into three variants of the disease clinical course: group 1 - mild (n=105), group 2 - moderate (n=90), and group 3 - severe (n=51) courses. The scrapings of the mucous membrane of the cervical canal and washings from the vagina were obtained before the start of antimicrobial therapy. Quantitative determination of pathogens was carried out by sowing the biological material on solid nutrient environment. Identification of detached bacteria was carried out according to Bergeys. Mollicutes (Ureaplasma spp., Mycoplasma hominis) were determined by the cultural-fermentative method using test systems and the polymerase chain reaction. Vaginal washings of 121 sick women were used for immunological studies. The levels of myeloperoxidase, lysozyme, human β-defensin-2, immunoglobulins M, A, G, secretory IgA, lactoferrin, C3-component of complement, secretory component, and tumor necrosis factor-α were determined. The reference group consisted of 23 clinically healthy women. The Statistica 12.0 program package for Windows was used, and a difference of p<0.05 was considered verifi able. Results. In the majority of patients with acute uncomplicated pyelonephritis (85.6% of cases), concomitant chronic pelvic inflammatory diseases (colpitis, salpingitis, underlying medical condition of cervix) were observed. Classical bacteria were more often detected in vaginal washings of patients of all groups, while mollicutes were detected in scrapings of the mucous membrane of the cervical canal, whereas the frequency of their detection increased with increase in the pyelonephritis severity. The analysis of indicators of local immunity determined disorders of mucosal immunity of the genital tract in the form of increased levels of myeloperoxidase, IgA, and IgG relative to reference values. The highest levels of lysozyme and myeloperoxidase were determined in patients with a severe course of acute uncomplicated pyelonephritis (by 5.3 and 3.6 times more, respectively), and the content of immunoglobulins A, M, and G in patients with a mild course of the disease (by 3.2, 3.1, and 4.0 times more, respectively). An increase in the level of tumor necrosis factor-α was detected in all patients, although no significant differences from controls were found in any group, but the highest median value was recorded in patients with severe pyelonephritis. In the group of patients with a severe clinical course, the lowest median was observed for β-defensin-2, which makes it impossible to inhibit the synthesis of TNF-α and, thus, supports the inflammatory process. Conclusions. It has been proven that in women with acute uncomplicated pyelonephritis, the severity of the course probably correlates with infection of the genital tract by mollicutes, mainly in association with classical bacteria and with level of mucosal immunity disorder. The obtained results of microbiological and immunological studies of biological material collected before the start of antibacterial therapy in patients with acute uncomplicated pyelonephritis proved the need for a mandatory examination by a gynecologist to ascertain concomitant chronic pelvic inflammatory diseases and establish infection with mollicutes to provide relevant etiotropic treatment. The above is the basis for adding the data regarding laboratory examination and treatment to the management protocols of patients with acute uncomplicated pyelonephritis.
急性无并发症肾盂肾炎不同病程育龄妇女生殖道微生物群及黏膜免疫紊乱
本研究探讨了急性无并发症肾盂肾炎伴慢性盆腔炎妇女生殖道的免疫-微生物相似性。除致病因性抗菌药物治疗外,纠正生殖道黏膜免疫状态对治疗本病和预防其复发至关重要。目的是调查不同临床病程的急性无并发症肾盂肾炎妇女生殖道污染和局部免疫状况。方法:246例育龄妇女急性无并发症肾盂肾炎,按病程分为3组:1组轻度病程(105例),2组中度病程(90例),3组重度病程(51例)。在开始抗菌治疗前,获得宫颈黏膜刮片和阴道冲洗。采用在固体营养环境下播种的方法对病原菌进行定量测定。根据Bergeys法对分离菌进行鉴定。采用培养-发酵法,采用检测系统和聚合酶链反应法测定分子(脲原体、人支原体)。121名患病妇女的阴道冲洗液用于免疫学研究。测定脊髓过氧化物酶、溶菌酶、人β-防御素-2、免疫球蛋白M、A、G、分泌IgA、乳铁蛋白、补体c3组分、分泌组分、肿瘤坏死因子-α的水平。参照组由23名临床健康妇女组成。采用Windows的Statistica 12.0程序包,差异0.05为可验证。结果。大多数急性无并发症肾盂肾炎患者(85.6%)伴有慢性盆腔炎(阴道炎、输卵管炎、宫颈基础疾病)。各组患者阴道洗液中检出经典细菌较多,宫颈粘膜刮擦中检出莫里特菌,且随肾盂肾炎严重程度的增加检出频率增加。局部免疫指标分析确定了生殖道粘膜免疫紊乱,表现为髓过氧化物酶、IgA和IgG水平相对于参考值升高。急性无并发症肾盂肾炎重症患者溶菌酶和髓过氧化物酶水平最高(分别高出5.3倍和3.6倍),免疫球蛋白a、M和G含量在轻度病程患者中最高(分别高出3.2倍、3.1倍和4.0倍)。所有患者均检测到肿瘤坏死因子-α水平升高,尽管与对照组相比,任何组均无显著差异,但严重肾盂肾炎患者的中位数最高。在临床病程严重的患者组中,β-防御素-2的中位数最低,这使得它无法抑制TNF-α的合成,从而支持炎症过程。结论。经证实,女性急性无并发症肾盂肾炎病程的严重程度可能与分子感染生殖道有关,主要与经典细菌和粘膜免疫紊乱程度有关。急性无并发症肾盂肾炎患者开始抗菌治疗前收集的生物材料的微生物学和免疫学研究结果证明,需要由妇科医生进行强制性检查,以确定是否伴有慢性盆腔炎,并确定是否有分子感染,以提供相关的致病因治疗。以上是将实验室检查和治疗资料加入急性无并发症肾盂肾炎患者管理方案的基础。
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Mikrobiolohichnyi zhurnal
Mikrobiolohichnyi zhurnal Medicine-Microbiology (medical)
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