In vitro models to study human gut-microbiota interactions: Applications, advances, and limitations.

In vitro models of the human gut help compensate for the limitations of animal models in studying the human gut-microbiota interaction and are indispensable in the clarification the mechanism of microbial action or in the high-throughput screening and functional evaluation of probiotics. The development of these models constitutes a rapidly developing field of research. From 2D1 to 3D2 and from simple to complex, several in vitro cell and tissue models have been developed and continuously improved. In this review, we categorized and summarized these models and described their development, applications, advances, and limitations by using specific examples. We also highlighted the best ways to select an appropriate in vitro model, and we also discussed which variables to consider when imitating microbial and human gut epithelial interactions.

Antioxidative Effect of Chlorella Pyrenoidosa Protein Hydrolysates and Their Application in Krill Oil-in-Water Emulsions.

Chlorella pyrenoidosa is an excellent source of protein, and in this research, we assessed the antioxidant and emulsifying effects of Chlorella protein hydrolysate (CPH) using neutral proteases and alkaline proteases, as well as the properties of CPH-derived krill oil-in-water (O/W) emulsions. The CPHs exhibited the ability to scavenge several kinds of free radicals, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), O2-, hydroxyl, and ABTS. Additionally, the CPHs (5 mg/mL) scavenged approximately 100% of the DPPH and ABTS. The CPHs showed similar emulsifying activities to Tween 20 and excellent foaming activities (max FS 74%), which helped to stabilize the krill oil-in-water emulsion. Less than 10 mg/mL CPHs was able to form fresh krill oil-in-water emulsions; moreover, the CPHs (5 mg/mL) in a krill O/W emulsion were homogenous, opaque, and stable for at least 30 days. Based on their inhibitory effects on the peroxide value (POV) and thiobarbituric acid reactive substances (TRABS), the CPHs were found to be able to inhibit lipid oxidation in both emulsifying systems and krill O/W emulsions. Thus, the CPHs could improve superoxide dismutase (SOD) activities by 5- or 10-fold and decrease the high reactive oxygen species (ROS) level caused by the addition of H2O2 in vitro. In conclusion, health-promoting CPHs could be applied in krill oil-in-water emulsions as both emulsifiers and antioxidants, which could help to improve the oxidative and physical stability of emulsions.

The Protective Effects of Ginseng Polysaccharides and Their Effective Subfraction against Dextran Sodium Sulfate-Induced Colitis.

Polysaccharides from Panax ginseng are natural carbohydrates with multiple activities. However, little was known about its functions on colitis. In this study, we aim to investigate the protective effects of ginseng polysaccharides and its effective subfraction on dextran sodium sulfate (DSS)-induced colitis. Water soluble ginseng polysaccharides (WGP) were obtained from dry ginseng root, then purified to neutral fraction (WGPN) and acidic fraction (WGPA) by ion exchange chromatography. An animal model was constructed with male Wistar rats, which were treated with a normal diet (con group), DSS (DSS group), WGP (WGP group), WGPN (WGPN group), and WGPA (WGPA group), respectively. Both WGP and WGPA alleviated the colitis symptoms and colon structure changes of colitis rats. They decreased the disease activity index (DAI) scores and improved colon health; reduced colon damage and recovered the intestinal barrier via regulating the tight-junction-related proteins (ZO-1 and Occludin); downregulated inflammatory cytokines (IL-1ß, IL-2, IL-6, and IL-17) and inhibited the TLR4/MyD88/NF-κB-signaling pathway in the colon; regulated the diversity and composition of gut microbiota, especially the relative abundance of Ruminococcus; enhanced the production of SCFAs. In conclusion, WGP exerted a protective effect against colitis with its acidic fraction (WGPA) as an effective fraction. The results support the utilization and investigation of ginseng polysaccharides as a potential intervention strategy for the prevention of colitis.

A. muciniphila Supplementation in Mice during Pregnancy and Lactation Affects the Maternal Intestinal Microenvironment.

During pregnancy and lactation, considerable factors that affect the maternal microbiome are associated with the advancement of numerous diseases, which can potentially affect offspring health. Probiotics have shown potential for the maintenance of microbiota homeostasis of mothers in this period. The specific objective of this study was to investigate whether the application of Akkermansia muciniphila (A. muciniphila) during pregnancy and lactation impacts maternal and offspring health. Here we show that dams fed with A. muciniphila is safe, enhances the intestinal barrier and alters gut microbiota composition and diversity at the end of lactation, including the significant enrichment of A. muciniphila and Ruminococcus_1 in offspring from probiotic-fed dams. However, compared with the control group, the fecal metabolites of the A. muciniphila group only changed slightly. Additionally, A. muciniphila supplementation did not significantly increase the abundance of A. muciniphila in the fecal microbiota of offspring mice. Compared with the control group, the fecal metabolic profile of three-week-old offspring of mice fed with A. muciniphila were significantly changed, containing the D-glutamine and D-glutamate metabolism pathways. These results provided evidence that A. muciniphila supplementation in mice during pregnancy and lactation is safe and seemed to have a more beneficial effect on dams. In the future, using probiotics to regulate maternal microbiomes during pregnancy and lactation could be shown to have a more lasting and beneficial effect.

The Structure Features and Improving Effects of Polysaccharide from Astragalus membranaceus on Antibiotic-Associated Diarrhea.

Astragalus membranaceus (Astragalus) is often used as a medical and food resource in China. The present study was designed to investigate the features and effects of polysaccharide from Astragalus membranaceus (WAP) on rats with antibiotic-associated diarrhea (AAD). WAP was mainly composed of glucose, galactose, arabinose and glacturonic acid, with glucan, arabinogalactan and RG-I regions, and it showed loosely irregular sheet conformation. WAP decreased the inflammatory cell infiltration of colon in AAD rats, increased propionate and butyrate production, improved metabolic levels, adjusted the diversity and composition of gut microbiota, increased the relative abundance of Pseudomonas, and decreased the relative abundance of Allobaculum and Coprococcus. In conclusion, WAP contained different types of polysaccharide regions and sheet three-dimensional conformation, while it ameliorated AAD by recovering the colon structure, adjusting the gut microbiota, and improving the SCFAs levels. The results can provide some data basis for natural products to alleviate the side effects related to antibiotics.

Effects of different substrates on low-temperature storage of fresh ginseng.

BACKGROUND: Fresh ginseng was buried in three types of sand with different moisture contents and three types of soil and then stored at 2 °C to determine the effects of these storage substrates on fresh ginseng. RESULTS: At a storage time of 200 days, ginseng stored in underforest soil softened the most slowly and had a significantly greater firmness compared to the other samples (P < 0.05). The amount of most ginsenosides changed after storage for most of the substrates. Samples stored in ginseng soil and biological fertilizer had the highest concentration of total saponin and ginseng polysaccharides, respectively. Fresh ginseng stored in medium-water content sand had a significantly lower polyphenol oxidase activity (P < 0.05). A significant difference was observed in the total concentration of nucleosides and nucleobases between the ginseng samples stored with and without substrates (P < 0.05). CONCLUSION: The data obtained in the present study suggest that the use of storage substrates is an optimal method for extending the shelf life of fresh ginseng without detrimental effects on its components. © 2019 Society of Chemical Industry.

[Effects of ginseng neutral polysaccharide on gut microbiota in antibiotic-associated diarrhea mice].

To investigate the effect of ginseng neutral polysaccharide on gut microbiota composition and diversity as well as the therapeutic effect for antibiotic associated diarrhea( AAD) in mice. The water-soluble ginseng neutral polysaccharide( WGPN) was purified from water-soluble ginseng polysaccharides( WGP) by DEAE-sepharose fast flow column,which was obtained from the roots of Panax ginseng. AAD mice were induced by gastric gavage with lincomycin hydrochloride,followed by administration of normal saline( natural recovery group,NR) or WGPN( WGPN group) for one week. Body weight changes,psychosis and diarrhea status were observed and assessed. 12 h after the last administration,histological observation of ileum and 16 S rRNA high throughput sequencing analysis of intestinal contents were conducted to identify the effects of WGPN on AAD mice. The results showed that WGPN could alleviate the symptoms of diarrhea in mice,decrease the inflammation and edema of ileum,and increase the length of intestinal villi. As compared to NR mice,WGPN could increase the relative abundance of Lactobacillus,and significantly decrease the relative abundance of Bacteroides,Streptococcus,Ochrobactrum and Pseudomonas at the genus level. In conclusion,WGPN could improve the gut microecology by recovering the ileum structure and improving the diversity and composition of the gut microbiota in AAD mice.

A Comparative Study on the Effects of Different Parts of Panax ginseng on the Immune Activity of Cyclophosphamide-Induced Immunosuppressed Mice.

The objective of the present study was to compare the effects of the immunological activity of various parts (root/stem/leaf/flower/seed) of five-year-old ginseng on the immune system of immunosuppressive mice. Immunosuppression was induced by cyclophosphamide (CTX) in the mouse model, whereas levamisole hydrochloride tablet (LTH) was used for the positive control group. We found that ginseng root (GRT), ginseng leaf (GLF), and ginseng flower (GFR) could relieve immunosuppression by increased viability of NK cells, enhanced immune organ index, improved cell-mediated immune response, increased content of CD4⁺ and ratio of CD4⁺/CD8⁺, and recovery of macrophage function, including carbon clearance, phagocytic rate, and phagocytic index, in immunodeficient mice. However, ginseng stem (GSM) and ginseng seed (GSD) could only enhance the thymus indices, carbon clearance, splenocyte proliferation, NK cell activities, and the level of IL-4 in immunosuppressed mice. In CTX-injected mice, GRT and GFR remarkably increased the protein expression of Nrf2, HO-1, NQO1, SOD1, SOD2, and CAT in the spleen. As expected, oral administration of GRT and GFR markedly enhanced the production of cytokines, such as IL-1ß, IL-4, IL-6, IFN-γ, and TNF-α, compared with the CTX-induced immunosuppressed mice, and GRT and GFR did this relatively better than GSM, GLF, and GSD. This study provides a theoretical basis for further study on different parts of ginseng.

Study on Antidepressant Activity of Pseudo-Ginsenoside HQ on Depression-Like Behavior in Mice.

Suppressive effects of ginsenoside Rh2 (Rh2), (24R)-pseudo-ginsenoside HQ (R-PHQ), and (24S)-pseudo-ginsenoside HQ (S-PHQ) against lipopolysaccharide (LPS)-induced depression-like behavior were evaluated using the forced swimming test (FST) and tail suspension test (TST) in mice. Pretreatment with Rh2, R-PHQ, and S-PHQ significantly decreased immobility time in FST and TST with clear dose-dependence, and significantly downregulated levels of serum tumor necrosis factor-α and interleukin-6, and upregulated superoxide dismutase activity in the hippocampus of LPS-challenged mice. Furthermore, R-PHQ and S-PHQ significantly increased the expression of the brain-derived neurotrophic factor (BDNF), tropomyosin-related kinase B (TrkB), sirtuin type 1 (Sirt1), and nuclear-related factor 2, and inhibited the phosphorylation of inhibitor of κB-α and nuclear factor-κB (NF-κB) in the hippocampus of LPS-challenged mice. Additionally, the antidepressant-like effect of R-PHQ was found related to the dopaminergic (DA), γ-aminobutyric acid (GABA)ergic, and noradrenaline systems, while the antidepressive effect of S-PHQ was involved in the DA and GABAergic systems. Taken together, these results suggested that Rh2, R-PHQ, and S-PHQ produced significant antidepressant-like effects, which may be related to the BDNF/TrkB and Sirt1/NF-κB signaling pathways.

Immunomodulatory Effects of (24R)-Pseudo-Ginsenoside HQ and (24S)-Pseudo-Ginsenoside HQ on Cyclophosphamide-Induced Immunosuppression and Their Anti-Tumor Effects Study.

(24R)-pseudo-ginsenoside HQ (R-PHQ) and (24S)-pseudo-ginsenoside HQ (S-PHQ) are the main metabolites of (20S)-ginsenoside Rh2 (Rh2) in vivo. In this study, we found that Rh2, R-PHQ, and S-PHQ upregulated the innate and adaptive immune response in cyclophosphamide (CTX) induced-immunocompromised mice as evidenced by the number of leukocytes, cellular immunity, and phagocytosis of macrophages. Spleen T-lymphocyte subpopulations and the serum cytokines level were also balanced in these immunosuppressed mice. Furthermore, co-administration with R-PHQ or S-PHQ did not compromise the antitumor activity of CTX in the hepatoma H22-bearing mice. Treatment with R-PHQ and S-PHQ clearly induced the apoptosis of tumor cells, significantly increased the expression of Bax, and remarkably inhibited the expression of Bcl-2 and vascular endothelial growth factor (VEGF) in H22 tumor tissues. The anti-tumor activity of R-PHQ and S-PHQ could be related to the promotion of tumor apoptosis and inhibition of angiogenesis and may involve the caspase and VEGF signaling pathways. This study provides a theoretical basis for further study on R-PHQ and S-PHQ.

[Effects of different processing methods on water soluble polysaccharide contents and monosaccharide compositions in Cervi Cornu Pantotrichum].

The water-soluble polysaccharides in antler velvet were extracted by water extraction and alcohol precipitation. The content was determined by a phenol-sulfuric acid colorimetry. After hydrolysis and derivatization, the various monosaccharide compositions of water-soluble polysaccharides from different parts of the antler velvet after different processing methods were analyzed by ultra high performance liquid chromatography (UPLC). The results showed that the water-soluble polysaccharide contents in wax slices, powder slices, gauze slices and bone slices of the boiled antler velvet were 1.74, 1.67, 1.03 and 1.13 g/kg, respectively, whereas those of the freeze-dried antler velvet were 2.77, 3.07, 1.22 and 3.20 g/kg, respectively. The water-soluble polysaccharide contents in in the same four parts of the antler velvet processed antler velvet without and with blood were 1.55, 1.78, 0.96, 0.77 g/kg, and 1.69, 1.64, 1.01, 1.31 g/kg, respectively. Eight monosaccharides (mannose, glucosamine, ribose, glucuronic acid, galacturonic acid, aminogalactose, glucose and galactose) were detected in the compositions of water-soluble polysaccharides in the antler velvet by using different processing methods. For the same part, the monosaccharide contents in the boiled antler velvet were lower than those in the freeze-dried antler velvet, and the monosaccharide contents (except those of glucosamine and aminogalactose) in the antler velvet processed without blood were lower than those in the antler velvet processed with blood. For different parts processed with the same method, the monosaccharide contents in wax and powder pieces were higher than those in gauze slices and bone pieces. This study provides a theoretical reference for velvet processing and product development.

Effects of Schisandra chinensis polysaccharides on rats with antibiotic-associated diarrhea.

Schisandra chinensis (S. chinensis), a traditional Chinese medicine (TCM), is comprised of polysaccharides as its main active component. In this study, water-soluble polysaccharides from S. chinensis (WSP) were extracted, and their effects on rats with antibiotic-associated diarrhea were investigated. The rats were treated with lincomycin hydrochloride, followed by saline (natural recovery [NR] group) or WSP (WSP group). The body weight and water intake of rats were analyzed, as well as histological data. Other experiments involving a measurement of cytokine levels, 16s rRNA sequencing analysis of caecal contents, and a measurement of the concentration of short chain fatty acids (SCFAs) were also performed. Compared to rats of the NR group, WSP decreased the infiltration of inflammatory cells into the ileum and colon; decreased the levels of IL-4, IL-17A and TNF-α; increased the serum IL-2 and IL-10 levels; increased the relative abundance of Blautia, Intestinibacter and Lachnospiraceae-UCG-008, but decreased the relative abundance of Ruminococcus-1, Ruminococcaceae-UCG-014 and Erysipelatoclostridium at the genus level; and significantly increased the contents of total SCFAs, acetate and propionate. In conclusion, WSP affected rats with antibiotic-associated diarrhea by improving the ultrastructure of the gut, adjusting cytokine levels and modifying the gut microbial community and SCFAs production.

Effects of Panax ginseng polysaccharides on the gut microbiota in mice with antibiotic-associated diarrhea.

Panax ginseng is a traditional medicinal plant used in most Asian countries to cure many diseases. The benefits of ginseng are due to its primary active component, polysaccharides. Gut microbiota dysbiosis is a worldwide problem associating with antibiotic use. The objective of this study was to investigate the effects of ginseng polysaccharides (WGP) on the diversity of the gut microbiota in mice with antibiotic-associated diarrhea. Compared to diarrhea mice, WGP significantly changed the composition and diversity of the gut microbiota. Specifically, WGP increased the relative abundance of the phylum Firmicutes and decreased the relative abundance of the phyla Bacteroidetes, Proteobacteria and Actinobacteria. At the genus level, WGP increased the relative abundance of Lactobacillus, Lactococcus, and Streptococcus, but decreased the relative abundance of Bacteroides. The key phylotype of beneficial bacteria in the gut microbiota that responded to WGP was Lactobacillus. In addition, WGP also reversed carbohydrate, amino acid and energy metabolism to normal levels, thereby promoting the recovery of the mucosal structure. Taken collectively, our results indicate that WGP altered the composition and diversity of the gut microbiota in mice with antibiotic-associated diarrhea, restored the gut microbiota, balanced metabolic processes, and promoted the recovery of the mucosa.