Kovač Urša, Žužek Zala, Raspor Dall'Olio Lucija, Pohar Katka, Ihan Alojz, Moškon Miha, Rozman Damjana, Starčič Erjavec Marjanca
Centre for Functional Genomics and Biochips, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia.
Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia.
Microorganisms. 2021 Apr 17;9(4):869. doi: 10.3390/microorganisms9040869.
Recent research has indicated that dysbiosis of the gut microbiota can lead to an altered circadian clock of the mammalian host. Herein we developed an original system that allows real-time circadian studies of human HepG2 hepatoma cells co-cultured with bacteria. The HepG2 cells with stably integrated firefly luciferase reporter under the control of promoter were co-cultured with strains isolated from human fecal samples from healthy individuals. The two strains differ in the phylogenetic group and the number of ExPEC virulence-associated genes: BJ17 has only two, and BJ23 has 15 of 23 tested. In the first 24 h, the BJ17 affected the HepG2 circadian clock more than BJ23. Cosinor analysis shows a statistically significant change in the amplitude of and and the phase advance of A high percentage of necrotic and apoptotic cells occurred at 72 h, while a correlation between the number of ExPEC genes and the influence on the HepG2 core clock gene expression was observed. Our study reveals that the genetic background is important for the effect on the mammalian circadian clock genes, indicating possible future use of probiotic strains to influence the host circadian clock.
最近的研究表明，肠道微生物群的失调会导致哺乳动物宿主的昼夜节律时钟发生改变。在此，我们开发了一种原始系统，该系统能够对与细菌共培养的人HepG2肝癌细胞进行实时昼夜节律研究。将在启动子控制下稳定整合萤火虫荧光素酶报告基因的HepG2细胞与从健康个体的人类粪便样本中分离出的菌株进行共培养。这两种菌株在系统发育组和ExPEC毒力相关基因的数量上有所不同：BJ17只有两个，而BJ23在23个测试基因中有15个。在最初的24小时内，BJ17对HepG2昼夜节律时钟的影响比BJ23更大。余弦分析显示，在中振幅和相位提前方面有统计学上的显著变化。在72小时时出现了高比例的坏死和凋亡细胞，同时观察到ExPEC基因数量与对HepG2核心时钟基因表达的影响之间存在相关性。我们的研究表明，菌株的遗传背景对哺乳动物昼夜节律时钟基因的影响很重要，这表明未来可能会使用益生菌菌株来影响宿主的昼夜节律时钟。
