| Objective:In recent years,the host-intestinal flora interaction has been the focus of attention.The interaction between host and microbial flora exists in all aspects of normal physiology,from nutrition to immune homeostasis.From obvious inflammatory bowel disease to complex diseases of external organs,there is growing evidence that there is a pathophysiological correlation between host and microorganism in patients with chronic kidney disease.Uremia affects the composition and metabolism of microorganisms.On the other hand,Important uremic toxins are derived from microbial metabolism.We studies the effects of Uremia patients intestinal flora on nutritional status and Micro-inflammation in rats,To provide a theoretical basis for the role of intestinal flora in chronic kidney disease.Method:1.Twenty-four SD male rats,which purchased from Animal Experiment Center of Dalian Medical University,were randomly divided into 3 groups: Control group(n=8),Uremic group(n=8),Healthy person group(n=8),8 Rats in each group.The Control group was free to eat and drank pure water,while the experimental group was free to eat and drank pure water with compound antibiotics(Ampicillin 1g/l,Gentamicin 240mg/l,Metronidazole 1g/l,Vancomycin 250 mg/l)for 3 weeks to establish the model of intestinal flora destruction in SD rats.2.Select 4 patients in CKD stage 5 of Non-dialysis from the Department of Nephrology,The second affiliated Hospital of Dalian Medical University and 4 healthy persons.Feces which collected from CKD5 patients and healthy persons(5g feces +100ml physiological saline were filtered after mixture,take the filtrate which according to 1.6ml/kg enema)were infused into the intestine of SD male rats of Uremic group and Healthy person group,and the specific enema methods were as follows:10% Chloral hydrate 400mg/kg by intraperitoneal injection into anesthesia of rats after supine position,3.5F rubber hose was inserted into the colon at 8-10 cm away from the anus,keep anus high about 5minutes.The Control group was not given special treatment.After that,the rats were fed regularly for 2 weeks.3.Then the rats were killed after anesthesia,taken blood and retained the fat around the epididymis and the kidney in the abdominal cavity.Albumin,serum creatinine,cholesterol,body fat ratio and body weight were detected by routine method.Serum interleukin-6(IL-6)and C-reactive protein(CRP)inflammatory markers were detected by ELISA method.Body fat ratio calculation: Weight of fat around epididymis and kidney in abdominal cavity/ body weight(g/100g).4.16 s r DNA amplifier sequencing technology was used to detect the intestinal flora of uremic patients,normal healthy persons,intestinal flora of SD rats after 3 weeks of oral antibiotics and intestinal flora of SD rats at 2 weeks after enema,and draw the flora abundance map.Result:1.The difference of intestinal flora between uremic patients and healthy people :The intestinal flora of uremic patients was significantly different from that of healthy people,mainly manifested by the increase of Firmicutes and Proteobacteria,and the decrease of actinomycete and Bacteroides.2.The effect of antibiotics on intestinal flora: Before the experiment,the body weight of the three groups of SD male rats were uniform with no significant difference.During the course of oral compound antibiotics,the intake of Oral antibiotic SD rats was lower than that in Control group.After taking compound antibiotics for 3 weeks,there was no significant difference in body weight between Uremic group and Healthy person group.(348.10±10.20 g VS 362.90±24.70 g,P > 0.05).After taking compound antibiotics for 3 weeks,the intestinal flora of SD rats in Uremic group and Healthy person group changed obviously,which showed that the diversity of intestinal flora was decreased obviously,which was mainly Proteobacteria and a small number of Firmicutes.3.The effects of fecal transplantation on nutritional status of rats: After enema by feces of uremic patients and healthy persons,the appetite and activity of SD rats in Uremic group decreased,and one died after enema in Uremic group,and 2weeks after enema,compared with healthy male SD rats,SD male rats in Uremic group had some weight loss,the difference is statistically significant(433.90±20.10 g VS466.70±12.60 g,P<0.05).Albumin in Uremic group is lower than that in Healthy person group,which the difference is statistically significant(20.36±7.50g/L VS22.87±5.44g/L,P<0.05).Body fat in Uremic group is lower than that in Healthy person group,which the difference is statistically significant(1.95±0.12g/L VS2.09±0.11g/L,P<0.05).Cholesterol in Uremic group is lower than that in Healthy person group,which the difference is statistically significant(1.96±0.39mmol/L VS2.54±0.73mmol/L,P<0.05).There was no significant difference in creatinine between Uremic group and Healthy person group(25.26±6.58umol/L VS 22.25±4.89umol/L,P >0.05).4.Effect of fecal transplantation on inflammatory indexes of rats:Two weeks after feces enema,interleukin-6 in Uremic group was higher than that in Healthy person group,which the difference is statistically significant(18.80±2.10pg/L VS14.30±1.80pg/L,P<0.05).C-reactive protein in Uremic group was higher than that in Healthy person group,which the difference is statistically significant(15.40±5.00mg/L VS 12.10±3.10mg/L,P<0.05).Conclusion: 1.Uremia has intestinal flora disorder.2.Fecal implantation of uremic patients can change the nutritional status of rats.3.Fecal implantation of uremic patients can affect the inflammatory response in rats.4.A new way is provided for the study of intestinal microflora through Flora transplantation after destroying the intestinal flora by antibiotics. |
PDF Full Text Request