| Objective and significance:Through in vitro and in vivo experiments,it was proved that Qingchanghuashi Decoction can promote the growth of probiotics,inhibit the growth of pathogenic bacteria,restore the micro-ecological balance of intestinal microflora in rats with ulcerative colitis,and inhibit intestinal mucosal damage.To provide basis and support for the treatment of ulcerative colitis with clean up the intestinal tract and remove moisture method.Research methods:1.The in vitro experiments:(1)Bifidobacterium and Lactobacillus rhamnosus bacillus liquids configured to have a concentration of 0.5 McFarland were inoculated into Bifidobacterium BS medium(without agar)and concentrations of 6.25%,12.5%,25%,50%Qingchanghuashi Decoction culture medium,respectively.After anaerobic incubation at 37℃ for 14h,16h,18h of Bifidobacteria and Lactobacillus rhamnosus 12h,14h,16h of Lactobacillus rhamnosus,then diluted 10 times and absorbance OD570 was measured to compare OD control and OD the Chinese medicine value was evaluated for its influence on the growth of Bifidobacterium and Lactobacillus rhamnosus in vitro.(2)Add 1ml of Qingchanghuashi Decoction extract with sterilized concentration of 3.8g/L(defined as 100%)to an equal volume of nutrient broth and add it to the first well of 24-well plate.Diluted to the 11th well by serial multiple dilution method.The 12th well was a positive control group without Qingchanghuashi Decoction,and the corresponding Chinese herbal medicine culture medium not inoculated with Escherichia coli was used as a blank group.Each well was added with Escherichia coli,Staphylococcus aureus,and Salmonella at a concentration of 0.5 McFarland.After incubation at 37℃ for 22 hours,the bacteriostatic effect was observed with a visual turbidimetric method,and an appropriate amount of the mixture was taken in each well to draw a line.The bactericidal effect was observed after incubation in a 37℃ incubator for 18 hours.2.Forty-eight male Sprague-Dawley rats were randomly divided into normal control group,model group,high-,middle-,and low-dose Qingjiehuashi Decoction group,and sulfasalazine group,a total of 6 groups.A rat model of ulcerative colitis was prepared by TNBS enema.After the model was established,high-,middle-,and low-concentrations of Qingchanghuashi Decoction were regularly administered orally and the rats were sacrificed after 12 days.The disease activity index(DAI)was assessed by assessing the body mass,stool characteristics,occult blood or gross bloody stools,visually observing the intestinal mucosa for colonic mucosal injury index scores,and observing the pathological changes of the intestinal mucosa under a microscope.Research findings:1、The 6.25%,12.5%,25%,and 50%concentrations of Qingchanghuashi Decoction all had a growth-promoting effect on bifidobacteria(P<0.01).Among them,25%concentration of Qingchanghuashi Decoction had the most obvious effect on promoting growth.2、The 25%and 50%concentrations of Qingchanghuashi Decoction had obvious growth-promoting effect on Lactobacillus rhamnosus at 12h and 14h(P<0.01),but it showed inhibitory effect at 16h.The 6.25%,12.5%concentration Qingchanghuashi Decoction had significant growth promoting effect on Lactobacillus rhamnosus at 12h(P<0.05).It had no obvious growth promoting effect at 14h(P>0.05),it inhibited at 16h.3.Qingchanghuashi Decoction has bacteriostasis and bactericidal effect on Escherichia coli and Chrysosporium.The MIC of the two drugs was 0.030g/L and 0.007g/L in turn,MBC were 0.119g/L and 0.238g/L in turn.And has bacteriostasis effects on Salmonella,the MIC was 0.030g/L.4.Qingchanghuashi Decoction can significantly reduce the DAI score and colonic mucosal injury index score in rats with ulcerative colitis,and there is a significant difference compared with the model group(P<0.05).Conclusion and significance:1.Qingchanghuashi Decoction can promote the growth of Bifidobacterium in vitro and has a two-way effect on the growth of Lactobacillus rhamnosus in vitro.2.The Qingchanghuashi Decoction has antibacterial and bactericidal effects on Escherichia coli and Staphylococcus aureus,and it has antibacterial effect on Salmonella.3.Qingchanghuashi Decoction can reduce the disease activity and colon mucosal injury in rats with ulcerative colitis. |
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