用时间生物学方法研究女性生殖道中微共生体过氧化氢酶的活性

Marina V. Nikolenko, E. M. Vaseva, Natalya V. Baryshnikova, O. I. Malishevskaya
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摘要

过氧化氢酶是一种含血红素的酶,属于破坏过氧化物化合物的保护因子。过氧化氢酶活性的存在是微生物的一项重要能力,可使其免受不利因素的影响,并适应大型生物的环境。过氧化氢酶和超氧化物歧化酶在病原体抵抗吞噬细胞依赖氧气的杀菌机制方面发挥着重要作用。本研究的目的是利用时间生物学方法研究正常菌群失调和念珠菌菌群失调情况下女性生殖道微生菌过氧化氢酶的活性。研究对象是从女性生殖道微生菌病中分离出来的临床分离株。过氧化氢酶活性是通过分光光度法测定的,以冬季 24 小时过氧化氢减少量为基础,间隔 3 小时。动态过氧化氢在 3-5 个实验重复中进行评估。在一些乳酸菌属中发现了不含血红素基团的过氧化氢酶--假过氧化氢酶。时间生物学方法揭示了所有微生物共生体的酶活性。从健康女性体内分离出的优势微生物群和关联微生物群,其过氧化氢酶活性的昼夜(24 小时)节律为清晨 5 点(р 0.05)。过氧化氢会自发分解或通过非酶催化剂分解,微生物会在这种条件下应对这种情况。在女性生殖道菌群失调的微共生菌中,通常会发现大量乳酸杆菌,在早晨(上午 8 点)和傍晚(晚上 8 点),过氧化氢酶活性的 12 小时和 8 小时谐波与尖峰相的超昼夜节律被记录下来。因此,所研究参数的节律在不同阴道不育程度下的贡献反映了病原体对生存条件的适应能力,可作为研究相关调节机制的基础。间期和振幅相位稳定性是通用的节律参数,用于评估患者的状况,与物种分配无关。
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Chronobiological approach to study microsymbiont catalase activity in female reproductive tract
Catalase is a heme-containing enzyme belonging to protection factors that destroys peroxide compounds. The presence of catalase activity is an important ability of microorganisms that allows them to be protected from unfavorable factors as well as adapt to macroorganism conditions. Catalase along with superoxide dismutase plays an important role in pathogen resistance to phagocyte oxygen-dependent bactericidal mechanisms. The aim of the study was to investigate microsymbiont catalase activity from female reproductive tract in normocenosis and candidiasis dysbiosis using the chronobiological approach. The study was conducted on clinical isolates, isolated from female reproductive tract microsymbiocenosis. The catalase activity was determined by spectrophotometry based on 24 hour-long hydrogen peroxide reduction with 3-hours interval in winter season. Dynamic hydrogen peroxide was assessed in 3–5 experiment replicates. In some Lactobacillus spp., catalase was found containing no heme group — pseudocatalase. Chronobiological approach allowed to reveal enzyme activity from all microsymbionts. The dominant and associative microbiota isolated from healthy females was characterized by circadian (24 hours) rhythms of catalase activity early in the morning — 5 a.m. (р 0.05). Hydrogen peroxide decomposes spontaneously or via non-enzymatic catalysts, and microorganisms cope with this situation under such conditions. In microsymbionts characteristic of female reproductive tract dysbiosis, and usually found in large numbers along with decreased Lactobacillus spp. ultradian rhythms with 12- and 8-hour harmonics of catalase activity with acrophase were recorded in the morning (8 a.m.) and evening hours (8 p.m.). The minimum values of enzyme production in all cultures were recorded at 12 p.m. and 5 p.m. Therefore, the contribution of the rhythm of the studied parameter at varying degree of vaginal sterility reflects the adaptive pathogen capabilities to the conditions of existence and can be the basis for studying related regulatory mechanisms. Mesor and amplitude phase stability are universal rhythmometric parameters used to evaluate patient’s condition independent of species assignment.
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