Codonopsis pilosula Polysaccharide Improved Spleen Deficiency in Mice by Modulating Gut Microbiota and Energy Related Metabolisms.

Codonopsis Radix (CR) is an important traditional Chinese medicine used for the treatment of spleen deficiency syndrome (SDS). Codonopsis pilosula polysaccharides (CPP) in CR are considered to be responsible for tonifying the spleen function; however, the mechanisms of the polysaccharides have remained unclear. This study aimed to investigate the treatment mechanisms of CPP in SDS mice using a combinational strategy of 16S rRNA gene sequencing and targeted metabolomics. Here, studies demonstrated that CPP had invigorating effect in vivo in Sennae Folium-induced SDS in mice by organ indexes, D-xylose determination, gastrointestinal hormones levels and goblet cells observation. Antibiotic treatment revealed that the intestinal microbiota was required for the invigorating spleen effect of CPP. Furthermore, gut microbiota analysis found that CPP significantly enriched probiotic Lactobacillus and decreased the abundance of some opportunistic pathogens, such as Enterococcus and Shigella. The metabolic profile analysis of the colonic content revealed that 25 chemicals were altered significantly by CPP, including amino acids, organic acids, fatty acids, carbohydrates and carnitine etc., which are mainly related to "energy conversion" related processes such as amino acids metabolism, tricarboxylic acid cycle, and nitrogen metabolism. Spearman's correlation assays displayed there were strong correlations among biochemical indicators-gut microbiota-metabolomics. In summary, these results provided a new perspective for CPP improving SDS by regulating energy metabolism related bacteria and pathways.

AMPK Activation of Flavonoids from Psidium guajava Leaves in L6 Rat Myoblast Cells and L02 Human Hepatic Cells.

OBJECTIVE: To isolate the hypoglycemic bioactive components from leaves of Psidium guajava and evaluate their AMP-activated protein kinase (AMPK) activities. METHODS: A variety of column chromatography was used for the isolation of compounds, and nuclear magnetic resonance (NMR) and mass spectrum (MS) were used for the structure identification of compounds. AMP-activated protein kinase (AMPK) activity of compounds obtained from leaves of Psidium guajava was evaluated in L6 rat myoblast cells and L02 human hepatic cells by western blot. RESULTS: Six principal flavonoids largely present in the leaves of Psidium guajava, quercetin (1), quercetin-3-O-α-L-arabinofuranoside (2), quercetin-3-O-α-L-arabinopyranoside (3), quercetin-3-O-ß-D-galactopyranoside (4), quercetin-3-O-ß-D-glucopyranoside (5), and quercetin-3-O-ß-D-xylopyranoside (6), were obtained and compound 1-6 exhibited significant activity on AMPK activation both in L6 cells and L02 cells (p < 0.01) compared with Control. In particular, the effects of quercetin on AMPK activation were extremely significant compared with Control (p < 0.001). CONCLUSIONS: These findings demonstrated that these flavonoids had potential for the activation of AMPK and hypoglycemic activity.

Three Inulin-Type Fructans from Codonopsis pilosula (Franch.) Nannf. Roots and Their Prebiotic Activity on Bifidobacterium longum.

Radix Codonopsis, derived from the roots of Codonopsis pilosula (Franch.) Nannf., Codonopsis pilosula (Franch.) Nannf. Var. modesta (Nannf.) L.T. Shen and Codonopsis tangshen Oliv., has been used as traditional Chinese medicine for improving poor gastrointestinal function, treating gastric ulcers and chronic gastritis in China. Inulin-type fructans are carbohydrates consisting mainly of ß (2→1) fructosyl-fructose links in chemical structure and exhibit a range of properties such as prebiotic activity, fat substitutes in low-calorie foods and disease-modifying effects. The prebiotic effects of inulin-type fructans are hypothesized to improve gastrointestinal function through alterations to gut microbiota composition and metabolism. In the present study, three inulin-type fructans with high degree of polymerization (DP = 16, 22, and 31) were isolated from the roots of Codonopsis pilosula (Franch.) Nannf. and their structures were confirmed by MALDI-TOF-MS, 1D- and 2D-NMR. The prebiotic activity of these fructans was evaluated by detecting growth stimulation on Bifidobacterium longum. The results demonstrated that three fructans at a concentration of 2.0 g/L exhibited significant growth stimulation on Bifidobacterium longum in a time-dependent manner (p < 0.01). The data indicated that inulin-type fructans in Radix Codonopsis could be used as potential prebiotics.

Structure Features and Anti-Gastric Ulcer Effects of Inulin-Type Fructan CP-A from the Roots of Codonopsis pilosula (Franch.) Nannf.

Radix Codonopsis has been used in traditional Chinese medicine for strengthening the immune system, improving poor gastrointestinal function, treating gastric ulcers and chronic gastritis and so on. In the present study, an inulin-type fructan CP-A was obtained from the roots of Codonopsis pilosula (Franch.) Nannf. and its structure was confirmed by MS and NMR as (2 → 1) linked-ß-d-fructofuranose. The protective effects of CP-A against ethanol-induced acute gastric ulcer in rats were intensively investigated. A Lacy assay demonstrated that CP-A-treated group (50 mg/kg) showed the gastric damage level 1, which was similar to the positive control group, while the model group exhibited the gastric damage level 3. The Guth assay demonstrated that the mucosa ulcer index for CP-A groups at the doses of 50 mg/kg and 25 mg/kg significantly decreased compared with that in the model group (p < 0.05). Meanwhile, CP-A significantly increased the activities of SOD and GSH-Px, and decreased the contents of MDA and NO, and the activity of MPO in gastric tissue in a dose-dependent manner (p < 0.05). The present research reported for the first time that inulin-type fructan CP-A were likely the potential component in Radix Codonopsis for treatment of acute gastric ulcers.

[AFLP and HPLC Fingerprints Analysis of Codonopsis Species from Original Areas and the Same Planting Base].

Objective: To investigate the influence of environment on the genetic structure and chemical ingredients of different Codonopsis species. Methods: Amplified fragment length polymorphism( AFLP) molecular markers were used to analyze the genetic polymorphism of Codonopsis species from 24 different populations,and high performance liquid chromatography( HPLC) fingerprints were performed to analyze the chemical ingredients of 10 batches of Codonopsis roots. Results: According to species classification,the 24 populations were categorized into three groups by UPGMA cluster method. The samples of Codonopsis pilosula var. modesta introduced from Wenxian( Gansu Province) and Jiuzhaigou County( Sichuan Province) were clustered into Group Ⅰ. The Codonopsis tangshen population introduced from Banqiao( Hubei Province) was clustered into Group Ⅱ. The populations from Codonopsis pilosula introduced to Shanxi Province GAP base,and the wild and cultured populations of Codonopsis pilosula collected from Shanxi Province were into Group Ⅲ. The similarity of HPLC fingerprints among three Codonopsis species introduced into the same environment was over 0. 8. Conclusion: Genetic differences of three Codonopsis species from original areas and Codonopsis species introduced into Shanxi Province were caused by their inter-species genetic characteristic. Meanwhile, the similarities of genetic backgrounds in Codonopsis species were related to geographic space, and chemical ingredients of Codonopsis were easily influenced by the cultivation environment.