Escherichia coli Affects Expression of Circadian Clock Genes in Human Hepatoma Cells

Abstract

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 PERIOD2 promoter were co-cultured with E. coli strains isolated from human fecal samples from healthy individuals. The two E. coli 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 E. coli BJ17 affected the HepG2 circadian clock more than BJ23. Cosinor analysis shows a statistically significant change in the amplitude of PER1 and 2 and the phase advance of PER3. 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 E. coli genetic background is important for the effect on the mammalian circadian clock genes, indicating possible future use of probiotic E. coli strains to influence the host circadian clock.

RadA, a Key Gene of the Circadian Rhythm of Escherichia coli

“…Circadian rhythms in pathogenic bacteria opens several research questions, including their involvement in host response and antibiotic resistance. Recently, Kovac et al showed that the genetic background of E. coli is important for the effect on mammalian circadian clock genes, indicating a possible future use of probiotic strains of E. coli to influence the host circadian clock [ 21 ]. Therefore, pathogenic bacteria could affect the circadian rhythm of the host immune system through their own rhythm.…”

The effects of Levilactobacillus brevis on the physiological parameters and gut microbiota composition of rats subjected to desynchronosis

“…showed conflicting results. Analysis of log-ratios in the context of feature rankings and sample metadata was carried out using the Qurro tool [ 23 ]. Log-ratios of the selected taxonomic features associated with time across samples are provided on Fig.…”

“…Probiotic intervention is an innovative approach for the normalization of CRs. This has been evidenced by studies showing a positive restorative effect of probiotics and prebiotics on sleep quality [ 18 , 19 ] and circadian desynchrony [ 20 – 23 ]. Since GABA's central role for sleep has been well documented, its implication in this relationship cannot be completely dismissed.…”

Determination of prophylactic and therapeutic effectiveness of probiotic strain Escherichia coli 39-SN

“…Its effectiveness was explained by replacing toxic bacteria with normalized intestinal microflora. Escherichia coli based preparations were mainly used to treat and prevent intestinal dysbiosis [ 10 – 12 ]. Probiotic microorganisms must be able to multiply in the gastrointestinal tract actively.…”

“…In the experiment, virulent cultures of E. coli 15, S. Typhimurium 371, were used to infect experimental animals. The most important requirement for the manufacture and control of biological products is the production flow chart of the drug prepared from the E. coli 39-SN strain [ 9 – 12 ]. For the preparation of the E. coli strain 39-SN, the main digest of Hottinger was used, containing 10% of liver extract, 0.4% of peptone, then distilled water was added so that the content of amine nitrogen was at least 200–250 mg/% [ 13 – 16 ].…”