| ObjectiveqPCR was applied to quantify the intestinal flora of UC rat acted by Periplaneta a mericana extract, analysing the variation of probiotics and pathogenic as to provide refer ence for further study. Methods(1) Ulcerative Colitis rat modeling groups: SD rats were 220, of which 200 rats were treated with DNCB joint acetic acid-induced rat model of ulcerative colitis. The medication administration group, normal control group and model control group were set respectively, 10 rats in each group, gender split even.(2) Preparation of the Plasmid standard: Specific primer of the under test flora was designed by the 16 S r DNA. The target segmentsconnected with the vehicle into E. Coli were culturedand identified, so as to preparethe plasmid standards. The standard curve were established and the appropriate concentrations were determined by a series of dilution.(3) Feces genomic DNA extraction and homogenization of concentration: to collect the feces of the rat with Ulcerative colitis and to extract the genomic DNA. Concentration was deter-mined by nucleic acid protein analyzer while adjusting the concentration of test sample for 20?g/m L.(4) Quantitative detection of intestinal flora: Fluorescent quantitative PCR method was applied to detect content changesof the total flora, Bacteroides, Bifidobacterium, Lactobacillus, Escherichia coli, Enterococcus in rat feces samples. Mean and standard deviation were calculated by ANOVA using SPSS 19.0 statistical software. Rusults(1) Test drug 06C-HYS enema dosing: Bifidobacterium: low-dose group compared with model group(9.17±0.51 VS 7.99±0.45, P=0); Lactobacillus: low-dose group, medium and high doses compared with the model group(8.40±0.18 VS 7.77±0.3, P=0.003),(8.48±0.52 VS 7.77±0.3, P=0.001),(8.33±0.09 VS 7.77±0.3, P=0.008); Escherichia coli: low and high dose groups compared with model group(7.34±0.03 VS 7.85±0.22, P=0.020),(7.22±0.13 VS 7.85±0.22, P=0.040). Enterococcus: high-dose group compared with model group(5.82±0.08 VS 7.77±0.3, P=0.003),had significant variation.(2) Test drug 06C-HYS gavage: Bacteroides: low and high dose groups compared with the model group(9.39±0.19 VS 8.96±0.28, P=0.019),(9.48±0.21 VS 8.96±0.28, P=0.005); Bifidobacterium: low, high-dose groups compared with the model group were(9.25±0.28 VS 8.17±0.32, P=0.001),(9.12±0.55 VS 8.17±0.32, P=0.004); Lactobacillus: low, medium and high dose groups were compared with the model group(8.75±0.35 VS 7.92±0.46, P=0.000),(8.57±0.31 VS 7.92±0.46, P=0.010),(8.42±0.39 VS 7.92±0.46, P=0.048), had significant variation.(3) Test drug TY-GZG enema administration: Escherichia coli: low-dose group compared with the model group(7.27±0.3 VS 7.85±0.2, P=0.021); Enterococcus: the high-dose group compared with the model group respectively(5.77±0.19 VS 6.3±0.08, P=0.033),(5.87±0.13 VS 6.3±0.08, P=0.002), had a significant reduction.(4) Test drug TY-GZG gavage: Bacteroides: low-dose group compared with the model group(9.39±0.32 VS 8.96±0.28, P=0.017); Lactobacillus: low-dose group compared with the model group(9.39±0.32 VS 8.96±0.28, P=0.017), had a significant increase.(5) Test drug 06C-HYS low and high dose enema group B / E values were compared with the control group(1.26±0.1 VS 1.02±0.08, P=0.000),(1.17±0.04 VS 1.02±0.08, P=0.004); test drug 06C-HYS low, high dose gavage group B / E values were compared with the control group(1.23±0.05 VS 1.03±0.04,P=0.004),(1.23±0.08 VS 1.03±0.04,P=0.004); test drug TY-GZG low dose enema administration group B / E value compared with the control group(1.16±0.05 VS 1.02±0.08, P=0.019), treatment group compared with the model group B / E was significantly increased. ConclusionsThe 06C-HYS and TY-GZG test drug of Periplaneta Americana extract makes the probiotic bacteria multiplied andthe pathogenic bacteria reduced in the UC rat intestinal, has ability to adjust the intestinal micro ecology, and balance the intestinal flora, which may have a certain relationship with UC treatment. |
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