Salvia miltiorrhiza polysaccharide and its related metabolite 5-methoxyindole-3-carboxaldehyde ameliorate experimental colitis by regulating Nrf2/Keap1 signaling pathway.

The roots of Salvia miltiorrhiza have been used in Traditional Chinese Medicine for thousands of years. However, tons of aerial parts of this plant are usually discarded in the production of roots preparation. To make better use of these plant resources, the polysaccharide isolated from the aerial part of S. miltiorrhiza was investigated for its potential protection against intestinal diseases. A pectic polysaccharide (SMAP-1) was isolated and characterized being composed of homogalacturonan as the main chain and rhamnogalacturonan type I as ramified region, with side chains including arabinans and possible arabinogalactan type I and II. SMAP-1 exhibited robust protective effects against dextran sodium sulfate (DSS)-induced colitis and restored colitis symptoms, colonic inflammation, and barrier functions. Anti-oxidative effects were also observed by up-regulating Nrf2/Keap1 signaling pathway. Additionally, the level of serum 5-methoxyindole-3-carboxaldehyde (5-MC) was restored by SMAP-1 identified in metabolomic analysis, being correlated with the aforementioned effects. Protection against oxidative stress on intestinal porcine enterocyte cells (IPEC-J2) by 5-MC was observed through the activation of Nrf2/Keap1 system, as also shown by SMAP-1. In conclusion, SMAP-1 could be a promising candidate for colitis prevention, and 5-MC could be the signal metabolite of SMAP-1 in protecting against oxidative stress in the intestine.

The comparison of preliminary structure and intestinal anti-inflammatory and anti-oxidative activities of polysaccharides from different root parts of Angelica sinensis (Oliv.) Diels.

ETHNOPHARMACOLOGICAL RELEVANCE: The root of Angelica sinensis, has been commonly used in gynecology for centuries, and is normally applied divided into different parts in various clinical applications. At present, the majority of existing studies focus on the volatile oil and ferulic acid extracted from different parts of A. sinensis, but there is a dearth of scientific information on its water-soluble polysaccharides. AIM OF THE STUDY: The structures of polysaccharides from plants, have been reported contributing to multiple pharmacological activities such as anti-oxidative, anti-inflammatory, anti-tumor and liver protection. Therefore, the focus of this study was on its anti-oxidative and anti-inflammatory activities in vitro, which would be based on the various polysaccharides with distinct structures obtained from different parts of the A. sinensis root. MATERIALS AND METHODS: Four parts of A. sinensis root were separated according to the Chinese Pharmacopoeia: head, body, tail and whole body. Crude polysaccharides were obtained by water extraction and ethanol precipitation method, and were further fractionated by DEAE Sepharose chromatographic column and gel filtration. The comparison of ASPs from different root parts were performed, including chemical compositions determined by colorimetric analysis, monosaccharide compositions measured by high performance liquid chromatography (HPLC), glycosidic linkage units determined by methylation and gas chromatography-mass spectrometry (GC-MS), organic functional groups determined by FT-IR, molecular weight (Mw) demarcated by gel permeation chromatography, and the viscosities and solubilities were measured according to method published in the previous report with minor modification. In vitro biological activities of APSs were compared on lipopolysaccharide (LPS)-induced inflammatory and oxidative stress models on IPEC-J2 cells. RESULTS: Four purified polysaccharides, ASP-H-AP, ASP-B-AP, ASP-T-AP and ASP-Hb-AP from the root of A. sinensis, were obtained, and consisted of various contents of protein and the polyphenol. They were possibly pectic polysaccharides with a long homogalacturonan region as the main backbone and ramified with rhamnogalacturonan I region, but they were differed by subregions and the relative contents of glycosidic units. The Mw of four pectic polysaccharides were ranged from 67.9-267.7 kDa. The infrared spectrum also showed that the four polysaccharide fractions contained the characteristic peaks of polysaccharides. Their distinct primary structure could lead to a variety of biological activities. In vitro biological assays suggested that four polysaccharide fractions can protect IPEC-J2 cells against the LPS-induced inflammation by down-regulating inflammation factors and related genes on IPEC-J2 cells. These polysaccharides also could alleviate oxidative stress on IPEC-J2 cells by up-regulating the gene and protein expressions of antioxidant enzymes. It was concluded that ASP-H-AP possessed better anti-inflammatory and anti-oxidative effects, while those of ASP-T-AP was relatively poor among the four polysaccharide fractions. CONCLUSION: All results indicated that the structure of pectic polysaccharides from different root parts of A. sinensis differed, which lead to their distinct anti-inflammatory and anti-oxidative activities. This may also be one of the factors why different parts of A. sinensis showed various pharmacological activities and applied independently in traditional use. In addition, it would be valuable for further studies on structure-activity relationship of polysaccharides obtained by different root parts of A. sinensis.

Restorative Effects of Inulin From Codonopsis pilosula on Intestinal Mucosal Immunity, Anti-Inflammatory Activity and Gut Microbiota of Immunosuppressed Mice.

An inulin (CPPF), isolated from a traditional Chinese herbal medicine Codonopsis pilosula, was characterized and demonstrated with potential prebiotic activity in vitro before. Based on its non-digested feature, the intestinal mucosa and microbiota modulatory effects in vivo on immunosuppressed mice were investigated after oral administration of 200, 100 and 50 mg/kg of CPPF for 7 days. It was demonstrated that the secretions of sIgA and mucin 2 (Muc2) in ileum were improved by CPPF, and the anti-inflammatory activities in different intestine parts were revealed. The intestine before colon could be the target active position of CPPF. As a potential prebiotic substance, a gut microbiota restorative effect was also presented by mainly modulating the relative abundance of Eubacteriales, including Oscillibacter, unidentified Ruminococcus and Lachnospiraceae after high-throughput pyrosequencing of V4 region of 16S rRNA analysis. All these results indicated that this main bioactive ingredient inulin from C. pilosula was a medicinal prebiotic with enhancing mucosal immune, anti-inflammatory and microbiota modulatory activities.

Pectic polysaccharide from Nelumbo nucifera leaves promotes intestinal antioxidant defense in vitro and in vivo.

In this study, the Nelumbo nucifera leaf polysaccharide (NNLP) was isolated by hot water extraction and ethanol precipitation. DEAE anion exchange chromatography and gel filtration were further performed to obtained the purified fraction NNLP-I-I, the molecular weight of which was 16.4 kDa. The monosaccharide composition analysis and linkage units determination showed that the fraction NNLP-I-I was a pectic polysaccharide. In addition, the NMR spectra analysis revealed that NNLP-I-I mainly consisted of a homogalacturonan backbone and rhamnogalacturonan I, containing a long HG region and short RG-I region, with AG-II and 1-3 linked rhamnose as side chains. The biological studies demonstrated that NNLP-I-I displayed antioxidant properties through mediating the Nrf2-regulated intestinal cellular antioxidant defense, which could protect cultured intestinal cells from oxidative stress and improve the intestinal function of aged mice.

New pectic polysaccharides from Codonopsis pilosula and Codonopsis tangshen: structural characterization and cellular antioxidant activities.

BACKGROUND: Codonopsis pilosula and Codonopsis tangshen are plants widely used in traditional Chinese medicine. Two pectic polysaccharides from the roots of C. pilosula and C. tangshen named as CPP-1 and CTP-1 were obtained by boiling water extraction and column chromatography. RESULTS: The core structures of both CPP-1 and CTP-1 comprise the long homogalacturonan region (HG) as the backbone and the rhamnogalacturonan I (RG-I) region as the side chains. CPP-1 has methyl esterified galacturonic acid units and a slightly lower molecular weight than CTP-1. Biological testing suggested that CPP-1 and CTP-1 can protect IPEC-J2 cells against the H2 O2 -induced oxidative stress by up-regulating nuclear factor-erythroid 2-related factor 2 and related genes in IPEC-J2 cells. The different antioxidative activities of polysaccharides from different source of C. pilosula may be result of differences in their structures. CONCLUSION: All of the results indicated that pectic polysaccharides CPP-1 and CTP-1 from different species of C. pilosula roots could be used as a potential natural antioxidant source. These findings will be valuable for further studies and new applications of pectin-containing health products. © 2021 Society of Chemical Industry.

Characterization of an antioxidant pectic polysaccharide from Platycodon grandiflorus.

Platycodonis Radix is widely used as homology of medicine and food in China; polysaccharides are thought to be one of its functional constituents. In this study, a pectic polysaccharide, PGP-I-I, was obtained from the root of the traditional medicine plant Platycodon grandiflorus through ion exchange chromatography and gel filtration. This was characterized being mainly composed of 1,5-α-L-arabinan and both arabinogalactan type I (AG-I) and II chains linked to rhamnogalacturonan I (RG-I) backbone linked to longer galacturonan chains. In vitro bioactivity study showed that PGP-I-I could restore the intestinal cellular antioxidant defense under the condition of hydrogen peroxide (H2O2) treatment through promoting the expressions of cellular antioxidant genes and protect against oxidative damages.

Characterization of inulin-type fructans from two species of Radix Codonopsis and their oxidative defense activation and prebiotic activities.

BACKGROUND: Codonopsis pilosula and C. tangshen are both plants widely used in traditional Chinese medicine. Polysaccharides, which are their primary active components, are thought to be important in their extensive use. In this study, two neutral polysaccharide fractions of C. pilosula (CPPN) and C. tangshen (CTPN) were obtained by fractionation on a DEAE-Sepharose column and characterized. RESULTS: It was confirmed that the neutral polymers CPPN and CTPN were ß-(2,1)-linked inulin-type fructans with non-reducing terminal glucose, and degree of polymerization (DP) of 19.6 and 25.2, respectively. The antioxidant and prebiotic activities in vitro were assayed based on IPEC-J2 cell lines and five strains of Lactobacillus. Results indicated that the effects of CPPN and CTPN were increased antioxidant defense in intestinal epithelial cells through enhanced cell viability, improved expression of total antioxidant capacity, glutathione peroxidase, superoxide dismutase and catalase, and reduced levels of malondialdehyde and lactic dehydrogenase. The prebiotic activity of CPPN and CTPN was demonstrated by the promoting effect on Lactobacillus proliferation in vitro. The different biological activities obtained between the two fractions are probably due to the different DP and thus molecular weights of CPPN and CTPN. CONCLUSION: The inulin fractions from C. pilosula and C. tangshen were natural sources of potential intestinal antioxidants as well as prebiotics, which will be valuable in further studies and new applications of inulin-containing health products. © 2020 Society of Chemical Industry.

A Polysaccharide Isolated from Codonopsis pilosula with Immunomodulation Effects Both In Vitro and In Vivo.

In this study, an acidic polysaccharide from Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen (WCP-I) and its main fragment, WCP-Ia, obtained after pectinase digestion, were structurally elucidated and found to consist of a rhamnogalacturonan I (RG-I) region containing both arabinogalactan type I (AG-I) and type II (AG-II) as sidechains. They both expressed immunomodulating activity against Peyer's patch cells. Endo-1,4-ß-galactanase degradation gave a decrease of interleukine 6 (IL-6) production compared with native WCP-I and WCP-Ia, but exo-α-l-arabinofuranosidase digestion showed no changes in activity. This demonstrated that the stimulation activity partly disappeared with removal of ß-d-(1→4)-galactan chains, proving that the AG-I side chain plays an important role in immunoregulation activity. WCP-Ia had a better promotion effect than WCP-I in vivo, shown through an increased spleen index, higher concentrations of IL-6, transforming growth factor-ß (TGF-ß), and tumor necrosis factor-α (TNF-α) in serum, and a slight increment in the secretory immunoglobulin A (sIgA) and CD4+/CD8+ T lymphocyte ratio. These results suggest that ß-d-(1→4)-galactan-containing chains in WCP-I play an essential role in the expression of immunomodulating activity. Combining all the results in this and previous studies, the intestinal immune system might be the target site of WCP-Ia.

Characterization of Inulin-Type Fructan from Platycodon grandiflorus and Study on Its Prebiotic and Immunomodulating Activity.

Platycodon grandiflorus is a plant widely used in traditional Chinese medicine, of which polysaccharides are reported to be the main components responsible for its bio-functions. In this work, the inulin-type fructan (PGF) was obtained by DEAE anion exchange chromatography from the water extracted from P. grandifloras. Characterization was performed with methanolysis, methylation, and NMR and the results showed that PGF is a ß-(2-1) linked fructan, with terminal glucose and with a degree of polymerization of 2⁻10. In order to study its biofunctions, the prebiotic and immunomodulation properties were assayed. We found that PGF exhibited good prebiotic activity, as shown by a promotion on six strains of lactobacillus proliferation. Additionally, the PGF also displayed direct immunomodulation on intestinal epithelial cells and stimulated the expressions of anti-inflammatory factors. These results indicated that the inulin from P. grandiflorus is a potential natural source of prebiotics as well as a potential intestinal immunomodulator, which will be valuable for further studies and new applications.

Therapeutic effect of Chinese patent medicine "Wuhuanghu" on porcine infectious pleuropneumonia and its acute and subchronic toxicity as well as evaluation of safety pharmacology.

Chinese patent medicines play an important role in veterinary clinical use. The aim of this study is to research the anti-infection effect of Chinese patent medicine "Wuhuanghu" for the treatment of porcine infectious pleuropneumonia and to evaluate the safety of "Wuhuanghu" in order to provide a comprehensive understanding of its toxicity. The anti-infection results showed that the treatment with "Wuhuanghu" could significantly inhibit pneumonia and decrement of the pneumonia in high, medium and low doses of "Wuhuanghu" groups were 70.97%, 61.29% and 58.06% respectively. The acute toxicity test showed that rats in the highest group (5000mg/kg) had no death and no abnormal response, suggesting the LD50 of "Wuhuanghu" was more than 5000mg/kg. The subchronic toxicity study showed that hematology indexes in all groups had no obvious differences; blood biochemical index, only albumin and total cholesterol in middle and low doses of "Wuhuanghu" groups were significantly decreased when compared with control group. The clinical pathology showed that the target organ of "Wuhuanghu" was liver. The safety pharmacology study indicated that "Wuhuanghu" had no side effects on rats. In conclusion, "Wuhuanghu" has therapeutic and protective effects to porcine infectious pleuropneumonia in a dose-dependent manner and "Wuhuanghu" is a safe veterinary medicine.

The reproductive toxicity of saponins isolated from Cortex Albiziae in female mice.

Saponin frsom Cortex Albiziae (SCA) are extensively used in the clinical treatment of tumor and depression. However, SCA may cause several adverse effects, including reproductive toxicity. The present study was designed to assess the mechanism by which SCA cause reproductive toxicity in female mice. The general reproductive toxicity testing was accomplished in female Kunming mice. The animals were divided into four groups: three groups that were treated by oral gavage with 135, 270, and 540 mg·kg(-1)·d(-1) of SCA prepared in physiological saline, respectively, and one vehicle control group that was treated with physiological saline only. The gestational toxicity tests were conducted at 540 mg·kg(-1)·d(-1). The general reproductive toxicity results showed that the pregnancy rate of the SCA-treated group decreased with the pregnancy rate being decreased by 70% at 540 mg·kg(-1)·d(-1). SCA elicited maternal toxicity in the ovary and the uterus, but no fetal toxicity or teratogenicity was observed. The rates of implantation in the early, middle, and late pregnancy were all decreased, with stillbirths and maternal deaths being observed. Histopathological changes showed that SCA adversely affected the ovary and the uterus. In conclusion, SCA-induced reproductive toxicity in female mice is most likely caused by its damage to the ovary and the uterus.

[Study on preparation process and stability of beta-cyclodextrin inclusion compound in volatile oil of Cinnamomum longepaniculatum leaves].

To study the optimum preparation process and stability of beta-cyclodextrin inclusion compound in volatile oil of Cinnamomum longepaniculatum leaves. The saturated aqueous solution method was adopted to prepare inclusion compounds for an orthogonal test. The inclusion compound productivity and the inclusion rate were taken as indexes for screening the inclusion processes. The inclusion effect was evaluated with the infrared spectrophotometry and TLC, and the stability under conditions of high temperature, high humidity and strong light was detected. Under optimum preparation conditions for inclusion, the ratio between volatile oil and beta-cyclodextrin was 1: 8 (mL: g), that between beta-cyclodextrin and water was 1: 15, the inclusion temperature was 40 degrees C, and the inclusion time was 3 h. The results of spectrophotometry and TLC showed that the optimum conditions can generate beta-cyclodextrin inclusion compound in volatile oil of C. longepaniculatum leaves with certain light resistance, thermo-stability and hygro-stability. Therefore the optimum inclusion process features simple operation and stable inclusion compounds.

Toxicological evaluation of neem (Azadirachta indica) oil: acute and subacute toxicity.

Neem (Azadirachta indica), popularly known as traditional medicine is a native plant in India. Neem oil is a vegetable oil derived from seeds or fruits of the neem tree through pressing or solvent extraction, and largely used in popular medicine to have antifungal, antibacterial, antimalarial, antiparasitic, anti-inflammatory, as well as immunemodulatory properties in different animal species. In the present study, acute and 28-day subacute toxicity tests were carried out. In the acute toxicity test, the LD50 values of neem oil were found to be 31.95g/kg. The subacute treatment with neem oil failed to change body weight gain, food and water consumption. Serum biochemistry analysis showed no significant differences in any of the parameters examined under the dose of 1600mg/kg/day. Histopathological exams showed that the target organs of neem oil were testicle, liver and kidneys up to the dose of 1600mg/kg/day.

Isolation and identification of the antibacterial active compound from petroleum ether extract of neem oil.

From a petroleum ether extract of neem oil (Azadirachta indica A. Juss) the new tetrahydrofuranyl diester 1 was isolated as an anti-bacterial constituent. 1 showed significant activities against three standard bacterial strains, including Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Salmonella enteritidis CMCC (B) 50041.

The preparation of neem oil microemulsion (Azadirachta indica) and the comparison of acaricidal time between neem oil microemulsion and other formulations in vitro.

The preparation of neem oil microemulsion and its acaricidal activity in vitro was developed in this study. In these systems, the mixture of Tween-80 and the sodium dodecyl benzene sulfonate (SDBS) (4:1, by weight) was used as compound surfactant; the mixture of compound surfactant and hexyl alcohol (4:1, by weight) was used as emulsifier system; the mixture of neem oil, emulsifier system and water (1:3.5:5.5, by weight) was used as neem oil microemulsion. All the mixtures were stired in 800 rpm for 15 min at 40 degrees C. The acaricidal activity was measured by the speed of kill. The whole lethal time value of 10% neem oil microemulsion was 192.50 min against Sarcoptes scabiei var. cuniculi larvae in vitro. The median lethal time value was 81.7463 min with the toxicity regression equations of Y=-6.0269+3.1514X. These results demonstrated that neem oil microemulsion was effective against Sarcoptes scabie var. cuniculi larvae in vitro.

[Hypoglyceminc effect of flavonoids from Prinsepia utilis on alloxan-induced diabetic mice].

OBJECTIVE: To evaluate the hypoglycemic effect of flavonoids from Prinsepia utilis Royle in alloxan-induced diabetic mice. METHODS: The hypoglycemic effects were investigated in alloxan-induced diabetic mice after oral administration of 300 mg/kg of flavonoids from Prinsepia utilis Royle for four weeks. The blood glucose (GLU), total cholesterol (TC), triglyceride (TG), very low density lipoprotein cholesterol (VLDL-C), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN) and body weight of mice were determined. RESULTS: Flavonoids from Prinsepia utilis Royle had influence on body weight increasing of diabetic mice in three weeks, but had no influence in the fourth week. Flavonoids from Prinsepia utilis Royle exhibited hypoglycemic effects. Among these fractions, flavonoids from Prinsepia utilis Royle significantly reduced GLU, TG and AST level in diabetic mice compared with model control group (P<0.01), markedly reduced VLDL-C, ALT and BUN level in diabetic mice compared with model control group (P<0.05), but had little influence on TC. CONCLUSION: Flavonoids from Prinsepia utilis Royle possess significant hypoglycemic activities, they can improve hypothepatia of diabetic mice and have protective effect on renal function of diabetic mice.