Intestinal Flora In Rats Research Articles

¹H-NMR-Based Metabonomics of the Protective Effect of Coptis chinensis and Berberine on Cinnabar-Induced Hepatotoxicity and Nephrotoxicity in Rats.

Coptis chinensis Franch has been used in Traditional Chinese Medicine (TCM) for treating infectious and inflammatory diseases for over two thousand years. Berberine (BN), an isoquinoline alkaloid, is the main component of Coptis chinensis. The pharmacological basis for its therapeutic effects, which include hepatoprotective effects on liver injuries, has been studied intensively, yet the therapy of liver injuries and underlying mechanism remain unclear. We investigated the detoxification mechanism of Coptis chinensis and berberine using metabolomics of urine and serum in the present study. After the treatment with Coptis chinensis and berberine, compared with the cinnabar group, Coptis chinensis and berberine can regulate the concentration of the endogenous metabolites. PLS-DA score plots demonstrated that the urine and serum metabolic profiles in rats of the Coptis chinensis and berberine groups were similar those of the control group, yet remarkably apart from the cinnabar group. The mechanism may be related to the endogenous metabolites including energy metabolism, amino acid metabolism and metabolism of intestinal flora in rats. Meanwhile, liver and kidney histopathology examinations and serum clinical chemistry analysis verified the experimental results of metabonomics.

16S rDNA analysis of the effect of fecal microbiota transplantation on pulmonary and intestinal flora.

This study aims to explore the effect of FMT on regulations of dysbacteriosis of pulmonary and intestinal flora in rats with 16S rDNA sequencing technology. A total of 27 SPF rats (3-4weeks old) were randomly divided into three groups: normal control group (K), model control group (MX), and fecal microbiota transplantation group (FMT); each group contained nine rats. The OTU values of the pulmonary and intestinal flora of the MX group decreased significantly compared with the normal control group. After FMT, the OTU value of pulmonary flora increased, while the value of OTU in intestinal flora declined. At the phylum level, FMT down-regulated Proteobacteria, Firmicutes, and Bacteroidetes in the pulmonary flora. At the genus level, FMT down-regulated Pseudomonas, Sphingobium, Lactobacillus, Rhizobium, and Acinetobacter, thus maintaining the balance of the pulmonary flora. Moreover, FMT could change the structure and diversity of the pulmonary and intestinal flora by positively regulating the pulmonary flora and negatively regulating intestinal flora. This study may provide a scientific basis for FMT treatment of respiratory diseases.

Effect of oral administration of Bacillus coagulans B37 and Bacillus pumilus B9 strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model

Aim:To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model.Materials and Methods:An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1) was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2) and (T3) groups received spore biomass of Bacillus coagulans B37 and Bacillus pumilus B9, respectively, suspended in sterilized skim milk at 8-9 log colony-forming units/ml plus basal diet for 28 days, while control group (T4) was supplied with clean water along with basal diet. There was a 14-day post-treatment period. A total of 288 fecal samples (8 fecal collections per rat) were collected at every 7-day interval starting from 0 to 49 days and subjected to the enumeration of the counts of coliforms and lactobacilli and Bacillus spores using respective agar media. In vitro acid and bile tolerance tests on both the strains were performed.Results:The rats those (T2 and T3) received either B. coagulans B37 or B. pumilus B9 spore along with non-fermented skim milk showed decrease (p<0.01) in fecal coliform counts and increase (p<0.05) in both fecal lactobacilli and Bacillus spore counts as compared to the control group (T4) and the group fed only skim milk (T1). In vitro study indicated that both the strains were found to survive at pH 2.0 and 3.0 even up to 3 h and tolerate bile up to 2.0% concentration even after 12 h of exposure.Conclusions:This study revealed that oral administration of either B. coagulans B37 or B. pumilus B9 strains might be useful in reducing coliform counts accompanied by concurrent increase in lactobacilli counts in the intestinal flora in rats.

The protective effect of Aplysin on ethanol‐induced liver damage via modulation of liver Kupffer cell function and altered intestinal flora in rats

Aplysin, a member of the bromine‐sesquiterpene compounds family, was mainly extracted from red alga Laurencia tristicha. This study aimed to explore the effects of Aplysin on rat liver, intestine, Kupffer cells and intestinal flora in rats exposed to alcohol. Rats were randomly divided into three groups: normal control group, ethanol‐model group, and Aplysin‐treated group. Except control group, rats were exposed to ethanol at dose of 6 g/kg/d for first four weeks, then 8 g/kg/d for last four weeks. One hour after that, Aplysin‐treated group was given oral administration of Aplysin at 150 mg/kg/bw on a daily basis. After 8 weeks, the rats were sacrificed and liver and intestine were collected for histological and biochemical assessments, plasma was collected for endotoxin level measurement with commercial kit, primary rat Kupffer cell cultures were established for measuring expression of CD14, TLR4 and NF‐kB using RT‐PCR and production of TNF‐α and IL‐1β in liver Kupffer cells using ELISA. Fecal samples were collected for quantitative analysis of intestinal floras. We found that Aplysin reduced the alcohol‐induced hepatic and intestinal histopathological changes and plasma endotoxin levels, and also suppressed the elevation of biomarkers for liver damage (AST, ALT, ALP, CHE and GGT). In primary Kupffer cell cultures, asplysin significantly decreased expression of CD14, TLR4, and NF‐kB as well as production of TNF‐α and IL‐1β. In addition, Aplysin significantly affected the changes in intestinal floras, including Escherichia coli, Enterococcus faecalis, Lactobacillus and Bifidobacterium. In conclusion, Aplysin exerted potent liver‐protective effect, which is associated with modulation of Kupffer cells function and altered intestinal flora.Support or Funding InformationThis work was supported by Grants from the National Nature Science Foundation of China (No.81573137) and Dietary Nutrition Research and Education Foundation of Danone Nutrition Center (No. DIC2014‐03)

The effect of ferrous-chelating hairtail peptides on iron deficiency and intestinal flora in rats.

Chelating agents, such as small peptides, can decrease free iron content and increase iron bioavailability. They may have promising therapeutic potential and may prevent the pro-oxidant effects of low molecular weight iron. Hairtail is a species of fish that is rich in easily digestible proteins. We extended this strategy for iron delivery by using an enzymatic hydrolysate of hairtail as the chelating agent and found that the ferrous-chelating hairtail peptides have anti-anaemic activity in Sprague-Dawley rats with anaemia. The anti-anaemic activity of ferrous-chelating hairtail peptides prepared by enzymatic hydrolysis of the hairtail and ferrous chelation was studied in rat models of iron deficiency anaemia. After the end of the 35 d experiment, we noted significant differences in haemoglobin, mean corpuscular volume, haemoglobin distribution width, and ferritin concentrations between those animals supplemented with ferrous-chelating hairtail peptides and FeSO4 and healthy animals. There were no negative side effects on the animals' growth or behaviour. There was no obvious inflammation in the intestinal mucosa lamina propria and no unbalance of intestinal flora. The novel ferrous-chelating hairtail peptides may be a suitable fortificant for improving iron-deficiency status. Our findings demonstrated that this multi-tracer technique has many applications in nutritional research. © 2015 Society of Chemical Industry.

Structure elucidation of in vivo and in vitro metabolites of mangiferin

The in vivo and in vitro metabolism of mangiferin was systematically investigated. Urine, plasma, feces, contents of intestinal tract and various organs were collected after oral administration of mangiferin to healthy rats at a dose of 200 mg/kg body weight. For comparison, mangiferin was also incubated in vitro with intestinal flora of rats. With the aid of a specific and sensitive liquid chromatography coupled with electrospray ionization tandem hybrid ion trap mass spectrometry (LC–ESI–IT–MS n ), a total of thirty-three metabolites of mangiferin were detected and their structures were tentatively elucidated on the basis of the characteristics of their precursor ions, product ions and chromatographic retention times. The biotransformation pathways of mangiferin involved deglycosylation, dehydroxylation, methylation, glycosylation, glucuronidation and sulfation.

Structural Elucidation of in vivo and in vitro Metabolites of Epiberberine in Rat and Its Liver and Intestines by Liquid Chromatography-Tandem Mass Spectrometry

The in vivo and in vitro metabolism of epiberberine was investigated using a highly specific and sensitive liquid chromatography–mass spectrometry (LC–MS/MS) method. In vivo samples including rat urine, feces, and plasma samples were collected individually after ingestion of 35 mg/kg epiberberine to healthy rats. In vitro samples were prepared by incubating epiberberine with homogenized liver and intestinal flora of rats, respectively. As a result, at least 17, 3 and 5 metabolites were found in rat urine, feces, and plasma, respectively. Additionally, 1 and 3 metabolites were found in the rat intestinal flora and homogenized liver incubation mixtures, respectively.

Intake of a high-fat diet alters intestinal flora in rats

Intake of a high-fat diet alters intestinal flora in rats

Colonic bacterial translocation as a possible factor in stress-worsening experimental stroke outcome

Stress is known to be one of the risk factors of stroke, but only a few experimental studies have examined the possible mechanisms by which prior stress may affect stroke outcome. In stroke patients, infections impede neurological recovery and increase morbidity as well as mortality. We previously reported that stress induces a bacterial translocation and that prior immobilization stress worsens experimental stroke outcome through mechanisms that involve inflammatory mediators such as release of proinflammatory cytokines and enzyme activation. We now investigate whether bacterial translocation from the intestinal flora of rats with stress before experimental ischemia is involved in stroke outcome. We used an experimental paradigm consisting of exposure of Fischer rats to repeated immobilization sessions before permanent middle cerebral artery occlusion (MCAO). The presence of bacteria and the levels and expression of different mediators involved in the bacterial translocation were analyzed. Our results indicate that stress before stroke is related to the presence of bacteria in different organs (mesenteric nodes, spleen, liver, and lung) after MCAO and increases inflammatory colonic parameters (such as cyclooxygenase-2, inducible nitric oxide synthase, and myeloperoxidase), but decreases colonic immunoglobulin A, and these results are correlated with colonic inflammation and bacterial translocation. Understanding the implication of bacterial translocation during stress-induced stroke worsening is of great potential clinical relevance, given the high incidence of infections after severe stroke and their main role in mortality and morbidity in stroke patients.

Liquid Chromatography-Electrospray Ionization Ion Trap Mass Spectrometry for Analysis of in Vivo and in Vitro Metabolites of Scopolamine in Rats

In vivo and in vitro metabolism of scopolamine is investigated using a highly specific and sensitive liquid chromatography-mass spectrometry (LC-MSn) method. Feces, urine, and plasma samples are collected individually after ingestion of 55 mg/kg scopolamine by healthy rats. Rat feces and urine samples are cleaned up by a liquid-liquid extraction and a solid-phase extraction procedure (C18 cartridges), respectively. Methanol is added to rat plasma samples to precipitate plasma proteins. Scopolamine is incubated with homogenized liver and intestinal flora of rats in vitro, respectively. The metabolites in the incubating solution are extracted with ethyl acetate. Then these pretreated samples are injected into a reversed-phase C18 column with mobile phase of methanol-ammonium acetate (2 mM, adjusted to pH 3.5 with formic acid) (70:30, v/v) and detected by an on-line MSn system. Identification and structural elucidation of the metabolites are performed by comparing their changes in molecular masses (DeltaM), retention-times and full scan MSn spectra with those of the parent drug. The results reveal that at least 8 metabolites (norscopine, scopine, tropic acid, aponorscopolamine, aposcopolamine, norscopolamine, hydroxyscopolamine, and hydroxyscopolamine N-oxide) and the parent drug exist in feces after administering 55 mg/kg scopolamine to healthy rats. Three new metabolites (tetrahydroxyscopolamine, trihydroxy-methoxyscopolamine, and dihydroxy-dimethoxyscopolamine) are identified in rat urine. Seven metabolites (norscopine, scopine, tropic acid, aponorscopolamine, aposcopolamine, norscopolamine, and hydroxyscopolamine) and the parent drug are detected in rat plasma. Only 1 hydrolyzed metabolite (scopine) is found in the rat intestinal flora incubation mixture, and 2 metabolites (aposcopolamine and norscopolamine) are identified in the homogenized liver incubation mixture.

Characterization of in vivo and in vitro Metabolic Pathway of Anisodamine by Liquid Chromatography‐Tandem Mass Spectrometry

In vivo and in vitro metabolisms of anisodamine were investigated using a highly specific and sensitive LC‐MSn method. Feces, urine, and plasma samples were collected individually after ingestion of 25 mg/kg anisodamine to healthy rats. Rat feces and urine samples were cleaned up by a liquid‐liquid extraction and a solid phase extraction procedure (C18 cartridges), respectively. Methanol was added to rat plasma samples to precipitate plasma proteins. Anisodamine was incubated with homogenized liver and intestinal flora of rats in vitro, respectively. The metabolites in the incubation solution were extracted with ethyl acetate. Then, these pretreated in vivo and in vitro samples, were injected into a reversed‐phase C18 column with mobile phase of methanol/0.01% triethylamine solution (adjusted to pH 3.5 with formic acid) (60:40, v/v) and detected by an on‐line MSn system. Identification and structural elucidation of the metabolites were performed by comparing their changes in molecular masses (ΔM), retention times and full scan MSn spectra with those of the parent drug. Fifteen new metabolites (aponoranisodamine, apoanisodamine, methoxyanisodamine, hydroxy‐methoxyanisodamine, trihydroxyanisodamine, dimethoxyanisodamine, dihydroxy‐methoxyanisodamine, tetrahydroxyanisodamine, hydroxy‐dimethoxyanisodamine, trihydroxy‐methoxyanisodamine, dihydroxy‐dimethoxyanisodamine, tetrahydroxy‐methoxyanisodamine, trihydroxy‐dimethoxyanisodamine, dihydroxy‐trimethoxyanisodamine, and hydroxy‐tetramethoxyanisodamine) were identified in rat urine after ingesting anisodamine. Seven metabolites (nor‐6β‐hydroxytropine, 6β‐hydroxytropine, tropic acid, aponoranisodamine, apoanisodamine, noranisodamine, and anisodamine N‐oxide) and the parent drug were detected in rat feces. Six metabolites (nor‐6β‐hydroxytropine, 6β‐hydroxytropine, tropic acid, apoanisodamine, hydroxyanisodamine, and anisodamine N‐oxide) and the parent drug are detected in rat plasma. Only apoanisodamine was detected in the homogenized liver incubation mixture. The hydrolyzed metabolites (6β‐hydroxytropine and tropic acid) and the dehydrated metabolite of anisodamine were found in the rat intestinal flora incubation mixture.

Structural elucidation of in vivo and in vitro metabolites of anisodine by liquid chromatography–tandem mass spectrometry

Liquid chromatography–electrospray ionization tandem mass spectrometry (LC–ESIMS n ) was employed to investigate the in vivo and in vitro metabolism of anisodine. Feces, urine and plasma samples were collected after ingestion of 20 mg anisodine to healthy rats. Feces and urine samples were cleaned up by liquid–liquid extraction and solid-phase extraction procedures (C18 cartridges), respectively. Methanol was added to plasma samples to precipitate plasma proteins. Anisodine was incubated with homogenized liver and intestinal flora of rats in vitro, respectively, followed by extraction with ethyl acetate. LC–MS n was used for the separation and identification of the metabolites using C18 column with mobile phase of methanol/0.01% triethylamine solution (2 mM, adjusted to pH 3.5 with formic acid) (60:40, v/v). The results revealed that five metabolites (norscopine, scopine, α-hydroxytropic acid, noranisodine and hydroxyanisodine) and the parent drug existed in feces. Three new metabolites (dimethoxyanisodine, tetrahydroxyanisodine and trihydroxy-methoxyanisodine) were identified in urine. Four metabolites (norscopine, scopine, hydroxyanisodine and anisodine N-oxide) and the parent drug were detected in plasma. Two hydrolyzed metabolites (scopine and α-hydroxytropic acid) were found in rat intestinal flora incubation mixture, and two metabolites (aponoranisodine and anisodine N-oxide) were identified in homogenized liver incubation mixture.

Effects of the lipid-saccharide complex and unsaponifiable matter from sunflowers on liver lipid metabolism and intestinal flora in rats.

The effects of the flower lipid-saccharide complex and unsaponifiable matter (1 g/kg of diet) from the sunflower on liver lipid metabolism and intestinal flora was studied in rats given cholesterol-enriched diets. After six weeks of feeding, the microsomal cholesterol concentration in the liver had been significantly reduced with the sunflower diet. The ratio of cholesterol/phospholipid was also reduced by the sunflower diet. The 3-hydroxy-3-methylglutaryl coenzyme A reductase activity of the sunflower groups was significantly lower than that of the control group. There was no significant difference in the cholesterol 7 alpha-hydroxylase activity, although this tended to increase with dietary sunflower consumption. The number of Bacillus in the cecum flora was significantly higher in the lipid-saccharide complex group than in the other groups, while Bifidobacterium and Eubacterium in the cecum flora was significantly higher in the unsaponifiable matter group when compared to the control group. These results suggest that the lipid-saccharide complex and unsaponifiable matter in the sunflower are related to liver cholesterol synthesis and intestinal flora.

Influence of diets low in protein or lysine on the cecal flora of rats with reference to cecal contents.

To determine the effect of a certain diet on the intestinal flora of rats, the cecal flora of rats fed a low protein or low lysine diet was examined. Total counts of bacteria in the cecal contents of rats given the four kinds of diet (a normal protein diet, a low protein diet, a normal lysine diet and a low lysine diet) were not significantly different. The counts of Streptococcus, Enterobacteriaceae, and Clostridium perfringens in the cecal contents from rats fed the low protein diet were significantly lower than those from rats fed the normal protein diet. The count of Lactobacillus in the cecal contents of the low protein group were significantly lower than those of the control. But no significant difference was found between the levels of most of the free amino acids in the cecal contents of the low lysine group and those of the control group.