Dynamic Variability Of Intestinal Flora And Endotoxin In Rats With Fulminant Hepatic Failure

BackgroundFulminant Hepatic Failure (FHF) is a serious disease in our country with 65-90% death rate. Infection is the commonest complication and one of factors that lead patients to death. Although antibiotics can cure infection, we can't ignore the fact that long-time and large-quantity use of them makes the increase of drug resistance. Moreover, at the same time that antibiotics wipe of pathogen, they also disturb the equilibrium of intestinal flora. Most of pathogens, leading to infections in the FFIF patients, come from intestinal flora, which physiologically keep in equilibrium in quantity. In FHF patients, intestinal flora is disordered, manifested by the decrease of Bifdobaterium and overgrowth of Enterobacteriacea. The disorder of intestinal flora playsan important role in bacterial translocation and infections. So, researches on the intestinal flora can uncover more about bacterial translocation and infections, and help us find Microbial Interference Therapies (MIT) to treat them. Now, using FHF rat models, we study on the dynamical viabilities of intestinal flora and endotoxin.Materials and Methods1. Experiment AnimalsForty male SD rats (Zhejiang Medical Animal Central), weighting 200-300g, are randomly divided into the control group and the FHF group. The control group (10 rats, Group A) was killed at the beginning of experiment while the FHF group was killed at 24 (12 rats, Group B) and 48 (18 rats, Group C) hours respectively after GlaN injection.2. Materials and MethodsD-galactosamine (GlaN) (Biochemical Institute, Chongqing Medical University) was freshly dissolved in physiological saline (1:4) and adjusted to pH 7.0. After fasting 12 hours, the rats of the group B and C were administered GlaN intraperitioneally two times at a dose of 1.4g/kg with 12 hours interval. Following culture mediums were used in our experiment: EMB EC TS EG NBGT LBS.3. Specimen Collection, Bacterial Culture and Endotoxin DetectingCollecting blood from right ventricle for measuring liver function parameters and endotoxin. Collecting blood from portal vein for measuring endotoxin. Collecting jejunum, ileum and feces in colons and diluting these samples for measuring endotoxin and bacterium culture. The bacteria were identified by morphology, Gram reaction, API Fermentation Tests. 4. Statistical AnalysisResults are expressed as Mean±SD. Statistical significance among the groups of parametric data was evaluated using x2 test, ANOVA or rank sum test. Differences were considered significant when possibility less then 0.05.Experiment Result and Discussion1. Death Rats All control rats survival. One rat in the Group B died between 16 and 24 hours after the second injection and the rest were depressed with the decrease of diet and reaction at the time of killing. In the Group C, eight rats died between 24 and 48 hours after the second injection, at the time of killing, one rat was excited with craziness and exasperation, four rats got some recover with turnover of spirit, disappearance of somnolence and increase of diet and activity, the rest were depressed with decrease of diet and reaction.2. Liver Function Parameters (see Diagram One)Diagram One: Liver Function ParametersLiver function parameters showed that the livers of the Group B and C were seriously injured with the decrease of TP ALB GLB and the increase of ALT AST AKP TBA TBiL. In the Group C, many rats died of liver failure while several rats of the survivals got some recovery at the time of killing. So, we thought that in the group C, liver function began to take a turn for better. 3. Intestinal Flora Analysis3.1 Enumeration of Enterobacteriacea (see Diagram Two)Diagram Two: Enumeration of Enterobacteriacea3.2 Enumeration of Lactobacillus (see Diagram Three)Diagram Three: Enumeration of Lactobacillus3.3 Enumeration of Enterococus (see Diagram Four)Diagram Four: Enumeration of enterococus3.4 Enumeration of Bacteroides (see Diagram Five)Dia...