| IntroductionWu Gu Chong is the drying larva of calliphoridae insects chrysomyia or other relatives insects, which is a traditional chinese medicine and is listed in many TCM classics like Compendium of Materia Medica, Yilin Zhuanyao and so on. The Wu Gu Chong has the effect on malnutrition in children, infantile malnutrition, kamtshng, pleen invigorating, removing heat and eliminating maluntrition. From the perspective of modern medicine, infantile malnutrition is a chronic digestive and metabolic dysfunction syndrome, whose main symptoms includes functional diarrhea, functional dyspepsia and so on.The dynamic changes of enterobacteria are closely related to the digestive and metabolic function of the organism. Based on such theory, this research created mice model with functional diarrhea to simulate the alteration of enterobacteria, made initial exploration into the effect of Wu Gu Chong on intestinal flora of mice.ObjectiveTo provide new drug development ideas in treating alteration of intestinal flora by studying the mediated effect of method on mice intestinal flora.Methods1. The effect of Wu Gu Chong on normal mouse.Preparation of insect powde: Adopting ancient method, fry the Wu Gu Chong until the color turned golden yellow. Crush and sift it.The mice were divided randomly into normal control group(physiological saline), high-dose group, medium-dose group and low-dose group. After 5 days of intragastric administration, excrements were collected to measure the amount of change in bifidobacterium, lactobacillus, enterococcus, enterobacter.2. The effect of Wu Gu Chong on mice with alteration of intestinal flora.Amount detection of intestinal flora: The mice models with functional diarrhea were created by gavaging folium sennae and divided randomly into normal control group(physiological saline), high-dose group, medium-dose group, low-dose group and spontaneous recovery group. After 5 days of intragastric administration, excrements were collected to measure the amount changes of bacillus bifidus, physiological saline, enterococcus and bacillar by plate count method.Small intestine tissue was taken for paraffin section, HE staining was used to observe the pathological changes of the tissues.3. Denaturing gradient gel electrophoresis:A denaturing gradient gel electrophoresis experiment was conducted to further study the changes of intestinal flora in mice before and after administration.4. Data statistic and analysisThe measurement data of each group were measured by mean plus or minus standard deviation(x±s), using SPSS 20.0 software for statistical analysis. presetting(P<0.05) as the standard of significant difference.Result1. The mediated effect of WuGuChong on normal mice.Preparation of insect powde: The color of the power was golden yellow and the texture was even. No impurities existed.Amount detection of intestinal flora: The result showed the increased on the bifidobacterium, lactobacillus, enterococcus and enterobacter in normal control group, but before and after the intragastric administration, there was no significant difference between this two phases(P>0.05); Compared with the control group, all the administration group has no significant difference showed in the amount of four kinds of bacteria(P>0.05); Before and after administration, the amount of four kinds of bacteria in low-dose group had increased but there was no significant difference(P>0.05); Both high-dose and medium-dose group showed a significant increase in the amount of bifidobacterium and lactobacillus. Enterococcus and enterobacter were the same, but their increase rate was less than that of bifidobacterium and lactobacillus(P<0.05).2. The mediated effect of Wu Gu Chong on mice with alteration of intestinal floraAmount detection of intestinal flora: Senna model mice showed decreased food intake, weight loss, listlessness and diarrhea; After modeling the amount of enterococcus and enterobacter increased significantly, but the number of bifidobacterium and lactobacillus decreased(P<0.05), indicating the presence of alteration of intestinal flora. In the normal control group, the amount of bifidobacterium, lactobacillus, enterococcus and enterobacter had increased after administration but there was no significant difference(P>0.05); Mice in low-dose group didn’t show great improvement of recovery, and no significant difference existed between the changes in the amount of four kinds of bacteria(P>0.05).The result was the same with natural recovery group(P>0.05); In the high-dose and medium-dose group, diarrhea symptoms lightened and finally returned to normal, bifidobacterium and lactobacillus had increased significantly, while the amount of enterococcus and enterobacter decreased(P<0.05), and the difference was significant with the spontaneous recovery group(P<0.05); All results in spontaneous recovery group also showed no significant change(P>0.05).The pathological slices of mouse intestine: The intestinal epithelial cells of model mice were destroyed after modeling and inflammatory infiltration appeared. After administration the intestinal mucosa epithelial cells in middle- dose group and high –dose group recovered and inflammatory cells decreased, showing the intestinal condition were turning to be normal; However, the recovery indication was not significant in low- dose group and spontaneous recovery group.3. Denaturing gradient gel electrophoresisInsect powder can increase the richness and diversity index of intestinal flora significantly in normal mice and functional diarrhea mice.Conclusion:The Wu Gu Chong had a certain mediated effect on intestinal flora of both normal mice and functional diarrhea mice, which can provide new scientific research ideas and development basis for regulating the gastrointestinal flora. |
