Combined Intestinal Metabolomics and Microbiota Analysis for Acute Endometritis Induced by Lipopolysaccharide in Mice

et al. 2023

Preprint

The tumor and tissue microbiota of human beings have recently been investigated. Gut permeability is known as a possible resource for the positive detection of tissue bacteria. Herein, we report that microbiotawere detected in high abundance in the hepatocytes of healthy rats, and that they were shared with the gut microbiota to an extent. We assessed male Sprague Dawley (SD) rats for the 16S ribosomal ribonucleic acid (rRNA) gene. After ratswere sacrificed by blood drainage from the portal vein, we extracted total deoxyribonucleic acid (DNA) from their ileal and colonic contents and liver tissues. The V3–V4 region of the 16S rRNA gene was amplified by polymerase chain reaction (PCR) and sequenced using an Illumina HiSeq 2500 platform. Sequences were assigned taxonomically by the SILVA database. We also detected bacterial lipopolysaccharide (LPS) and lipoteichoic acid (LTA) in situ using immunofluorescence (IF) and western blotting and the 16S rRNA gene using fluorescent in situ hybridization (FISH). In the livers of six rats, we detected 54,867.50 ± 6450.03 effective tags of the 16S rRNA gene and clustered them into 1003 kinds of operational taxonomic units (OTUs; 805.67 ± 70.14, 729–893). Individuals showed conservation of bacterial richness, abundance, and evenness. LPS and the 16S rRNA gene were detected in the nuclei of hepatocytes. The main function composition of the genomes of annotated bacteria was correlated with metabolism (79.92 ± 0.24%). Gram negativity was about 1.6 times higher than gram positivity. The liver microbiome was shared with both the small and large intestines, but showed significantly higher richness and evenness than the gut microbiome, and the β-diversity results showed that the liver microbiomeexhibited significantly higher similarity than the small and large intestines (P < 0.05). Our results suggest that the bacteria in the liver microbiome are hidden intracellular inhabitants in healthy rat livers.

Section: Discussionmentioning

confidence: 99%

Liver microbiome in healthy rats: the hidden inhabitants of hepatocytes

et al. 2023

Preprint

“…Findings have shown that various disorders in 9 Cellular Microbiology nearly every organ and system are associated with changes in the gut microbiome (samples primarily from feces) [52][53][54][55][56][57][58][59][60][61][62][63][64][65][66][67][68], and this has established the connection between the gut microbiome and diseases. It was suspected that gut microbiota and their metabolites, such as LPS, enter circulation (such as in bacteremia and toxemia) and result in organ injury [69,70]. However, whether the disease-associated changes in gut microbiota occur before or after disease development cannot be determined.…”

Section: Discussionmentioning

confidence: 99%

Liver Microbiome in Healthy Rats: The Hidden Inhabitants of Hepatocytes

et al. 2023

Cellular Microbiology

The tumor and tissue microbiota of human beings have recently been investigated. Gut permeability is known as a possible resource for the positive detection of tissue bacteria. Herein, we report that microbiota were detected in high abundance in the hepatocytes of healthy rats and that they were shared with the gut microbiota to an extent. We assessed male Sprague Dawley (SD) rats for the 16S ribosomal ribonucleic acid (rRNA) gene. After the rats were sacrificed by blood drainage from the portal vein, we extracted total deoxyribonucleic acid (DNA) from their ileal and colonic contents and liver tissues. The V3–V4 region of the 16S rRNA gene was amplified by polymerase chain reaction (PCR) and sequenced using an Illumina HiSeq 2500 platform. Sequences were assigned taxonomically by the SILVA database. We also detected bacterial lipopolysaccharide (LPS) and lipoteichoic acid (LTA) in situ using immunofluorescence (IF) and western blotting and the 16S rRNA gene using fluorescent in situ hybridization (FISH). In the livers of six rats, we detected 54,867.50 ± 6450.03 effective tags of the 16S rRNA gene and clustered them into 1003 kinds of operational taxonomic units (OTUs; 805.67 ± 70.14 , 729–893). Rats showed conservation of bacterial richness, abundance, and evenness. LPS and the 16S rRNA gene were detected in the nuclei of hepatocytes. The main function composition of the genomes of annotated bacteria was correlated with metabolism ( 79.92 ± 0.24 % ). Gram negativity was about 1.6 times higher than gram positivity. The liver microbiome was shared with both the small and large intestines but showed significantly higher richness and evenness than the gut microbiome, and the β-diversity results showed that the liver microbiome exhibited significantly higher similarity than the small and large intestines ( P < 0.05 ). Our results suggest that the bacteria in the liver microbiome are hidden intracellular inhabitants in healthy rat livers.

“…Findings have shown that various disorders in nearly every organ and system are associated with changes in the gut microbiota (samples primarily from feces) [53][54][55][56][57][58][59][60][61][62][63][64][65][66][67][68][69], and this has established the connection between the gut microbiota and diseases. It was suspected that gut microbiota and their metabolites, such as LPS, enter the circulation (such as in bacteremia and toxemia) and result in organ injury [70,71]. However, whether the disease-associated changes in gut microbiota occur before or after disease development cannot be determined; (2) Tissue microbiota in disease.…”

Section: Discussionmentioning

confidence: 99%

Liver microbiota in healthy rats: the hidden inhabitants of hepatocytes

et al. 2022

Preprint

The tumor and tissue microbiomes of human beings have recently been investigated. Gut permeability is known as a possible resource for the positive detection of tissue bacteria. Herein, we report that bacteria were detected in high abundance in the hepatocytes of healthy rats, and that they were shared with the gut microbiota to an extent. We assessed male Sprague Dawley (SD) rats for the 16S ribosomal ribonucleic acid (rRNA) gene. After rats were sacrificed by blood drainage from the portal vein, we extracted total deoxyribonucleic acid (DNA) from their ileal and colonic contents and liver tissues. The V3–V4 region of the 16S rRNA gene was amplified by polymerase chain reaction (PCR) and sequenced using an Illumina HiSeq 2500 platform. Sequences were assigned taxonomically by the SILVA database. We also detected bacterial lipopolysaccharide (LPS) and lipoteichoic acid (LTA) in situ using immunofluorescence (IF) and western blotting and the 16S rRNA gene using fluorescent in situ hybridization (FISH). In the livers of six rats, we detected 54,867.50 ± 6450.03 effective tags of the 16S rRNA gene and clustered them into 1003 kinds of operational taxonomic units (OTUs; 805.67 ± 70.14, 729–893). Individuals showed conservation of bacterial richness, abundance, and evenness. LPS and 16S rRNA gene were detected in the nuclei of hepatocytes. The main function composition of the genomes of the annotated bacteria was correlated with metabolism (79.92 ± 0.24%). Gram negativity was about 1.6 times higher than gram positivity. Liver bacteria were shared with both the small and large intestines but showed significantly higher richness, evenness, and β-diversity results showed that the liver microbiota exhibited significantly higher similarity than the small and large intestines (P < 0.05). The liver microbiota were hidden intracellular inhabitants in healthy rat livers and were shared with the gut microbiota.