| Human intestinal flora colonize in the human intestinal tract from birth, which may be associated with many complex diseases according to the perspective of a combination of taxonomy and functionality. Studies have shown that type 2 diabetes(T2D) is a heterogeneous and multifactorial disease that is affected by many different genetic and environmental factors and intestinal flora. Meanwhile, traditional C hinese medicine inevitably interacts with intestinal flora after oral taking. The flower of Coptis chinensis Franch. was used in this study to explore the role of intestinal microflora during the development of type 2 diabetes. The changes in the structure and type of intestinal flora were studied pre-treatment and post-treatment of the extracting solution of the flower of Coptis chinensis Franch..The studies are as following:1. Hydrochloric acid- methanol(1:100) was used to extract the active ingredients of 1g flower of Coptis chinensis Franch.. UV spectrophotometer was used to measure the content of total alkaloids. HPLC was used to measure the content of berberine. The total content of alkaloid in the flower of Coptis chinensis Franch. was 1.67%. The content of berberine was 0.86%.2. 50.0g flower of Coptis chinensis Franch. was weighed and extracted by ultrapure water. The extracting so lut io n was concentrated to 100 mL, obtaining 0.5g/mL stock solution. After adding 25%, 20%, 10%, 7.5%, 5%, 2.5% of the extracting so lut io n of the flower of Coptis chinensis Franch.(v/v) into MRS broth, Enterococcus broth, and EE broth, respectively, there was a different inhib it io n on the growth of four kinds of bacteria. The growth of Escherichia coli and Enterococcus faecalis was inhibited by treatment of various concentrations. However, Lactobacillus rhamnosus and Lactobacillus showed a slight proliferation in the culture medium containing 2.5% of the extracting solution of the flower of Coptis chinensis Franch..3. After feeding with high- fat and high-sugar diet for 6 weeks, SD rats were intraperitoneally injected with a small dose of STZ(30mg/K g). The successful T2 D model was characterized by polydipsia, polyuria, more food and weight loss of "a little more than three disease" and impaired glucose tolerance. Blood glucose level in treatment groups and control model group increased, but the increased trend of treatment groups was lower than that in control model group. Although the rising values of the medium-dose group and the low-dose group were lower than that of the control model group, the difference was not significant. There was a significant difference in the content of TG, CHO between high-dose group and control model group while there was no significant difference among other groups. Compared with treatment groups, control model group and normal group, the difference in the content of HDL-C, MDA, SOD and GSH-Px was not significant.4, The results of denaturing gradient gel electrophoresis(DGGE) showed that in type 2 diabetic rats, the number of Bacteroidetes significantly decreased while the number of Lactobacillus, Lactobacillus bulgaricus, Lactobacillus rhamnosus, Lactobacillus acidophilus and Lactobacillus casei increased. After treatment, the number of Blautia glucerasea, Lactobacillus animalis, Pseudomonas fluorescens etc. declined. Compared with other groups, DNA fingerprint analysis showed that the bacterial abundance in control model group was the highest. Princ ipa l component a na lysis showed that there was a significant difference a mo ng treatment groups, control model group and normal control group while there was also a significant difference between treatment groups and control mode l group. C luster a na lys is showed that the similarity between the model group and the normal control group was 0.36. The similarity between the treatment groups and the mode l control group was only 0.5.It can be seen that in vitro culture experiment of bacteria, except for 2.5% of the extracting so lut ion of the flower of Coptis chinensis Franch. could slightly promot the growth of Lactobacillus rhamnosus and Lactobacillus, the different conce ntrat io ns inhibited the growth of Escherichia coli and Enterococcus faecalis, indicating that microbial growth balance was broken. After treatme nt, blood glucose level and the conte nts o f TG, C HO in high, medium a nd low- dose gro ups red uced, whic h ind icated t hat the extracting so lut io n of the flower of Coptis chinensis Franch. had a therapeut ica l effect on diabetes. After DGGE a na lys is, the structure of the intestinal microbiota in treatment groups and model control group significantly changed. There was a different decline in the number of Lactobacillus and Lactobacillus rhamnosus. Therefore, the flower of Coptis chinensis Franch. probably played their therapeutic role by acting on intestinal microbiota. |
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