Effect of aqueous extract of Millettia speciosa Champ on intestinal health maintenance and immune enhancement of Cyprinus carpio.

Millettia speciosa Champ (MSP) is a natural Chinese herb that improves gastrointestinal health and enhances animal immunity. An 8-week feeding trial with different MSP levels (0, 150, 300, and 600 mg/kg) was conducted to evaluate the promotive effects of MSP in Cyprinus carpio. Results indicate that MSP improved intestinal immunity to some extent evidenced by the immuno-antioxidant parameters and the 16S rRNA in the Illumina MiSeq platform. With the analysis of transcriptome sequencing, 4685 differentially expressed genes (DEGs) were identified, including 2149 up-regulated and 2536 down-regulated. According to the GO and KEGG enrichments, DEGs were mainly involved in the immune system. Transcriptional expression of the NOD-like signaling pathway and key genes retrieved from the transcriptome database confirmed that innate immunity was improved in response to dietary MSP administration. Therefore, MSP could be used as a feed supplement that enhances immunity. This may provide insight into Chinese herb additive application in aquaculture production.

Millettia speciosa Champ cellulose-based hydrogel as a novel delivery system for Lactobacillus paracasei: Its relationship to structure, encapsulation and controlled release.

We report for the first time the usage of Millettia speciosa Champ cellulose (MSCC) and carboxymethylcellulose (MSCCMC) for the fabrication of 3D-network hydrogel as delivery system for probiotics. The structural features, swelling behavior and pH-responsiveness of MSCC-MSCCMC hydrogels and their encapsulation and controlled-release behavior for Lactobacillus paracasei BY2 (L. paracasei BY2) were mainly studied. Structural analyses demonstrated that MSCC-MSCCMC hydrogels with porous and network structures were successfully synthesized through the crosslinking of -OH groups between MSCC and MSCCMC molecules. An increasing concentration of MSCCMC significantly improved the pH-responsiveness and swelling ability of the MSCC-MSCCMC hydrogel toward neutral solvent. Besides, the encapsulation efficiency (50.38-88.91 %) and release (42.88-92.86 %) of L. paracasei BY2 were positively correlated with the concentration of MSCCMC. The higher the encapsulation efficiency was, the higher the release in the target intestine. However, due to the existence of bile salts, controlled-release behavior decreased the survivor rate and physiological state (degrading cholesterol) of encapsulating L. paracasei BY2. Even so, the number of viable cells encapsulated by hydrogels still reached the minimum effective concentration in the target intestine. This study provides an available reference for the practical application of hydrogels fabricated from the cellulose of the Millettia speciosa Champ plant for probiotic delivery.

Purification, structural elucidation and biological activities of exopolysaccharide produced by the endophytic Penicillium javanicum from Millettia speciosa Champ.

An acid polysaccharide, named HP, was produced by endophytic Penicillium javanicum MSC-R1 isolated from southern medicine Millettia speciosa Champ. The molecular weight of HP was 37.8 kDa and consisted of Ara f, Galр, Glcр, Manр, and GlcрA with a molar ratio of 1.09: 3.47: 68.48: 16.59: 8.85. The glycosidic linkage of HP was proven to be →3, 4)-α-D-Glcр-(1→6)-α-D-Manр-(1→, →3, 4)-α-D-Glcр-(1→4)-α-D-Glcр-(1→, →3), →6)-α-D-Manр-(1→4)-α-D-Glcр-(1→, →3), ß-D-Galр-(1→3)-α-D-Glcр-(1→, →4), →5)-α-L-Ara f -(1→3)-α-D-Glcр-(1→, →4), →6)-α-D-Manр-(1→4)-α-D-GlcAр-(1→ and →4)-α-D-GlcAр-(1→4)-α-D-Glcр-(1→, →3). Additionally, 250 µg/mL of HP possessed nontoxicity to RAW 264.7 cells and exhibited anti-inflammation activity. HP could significantly restrain the amount of tumor necrosis factor-α, interleukin-6 and NO release in RAW264.7, which property is possibly associated with its abundant glucosidic linkage. These results indicated that HP could be regarded as a ponderable ingredient for the health-beneficial functional foods.

Extraction and characterization of a functional protein from Millettia speciosa Champ. leaf.

Natural plant-derived protein with excellent bioactivities has attracted much attention so a functional protein with molecular weight of 15.2 kDa was extracted from Millettia speciosa Champ. leaf for the first time. Under the pH of 12.0, solid-liquid ratio of 1:40 (w/v), extraction time of 2.0 h, and extraction temperature of 50 °C, the highest extracting efficiency (79.25 ± 0.78%) of the Millettia speciosa Champ. leaf protein (MLP) was achieved. The main structure of MLP contained ß-fold and ß-corner by Fourier transform infrared spectroscopy (FTIR) and Circular dichroism (CD) spectra analysis. Additionally, MLP was predominant with glutamic acid, aspartic acid, and leucine, which could be considered as a high quality natural protein. MLP showed great water holding capacity (WHC), oil absorption capacity (OAC), as well as emulsifying and foaming properties. Simultaneously, MLP exhibited considerable antioxidant activity. These results suggested that MLP could be utilised as a promising ingredient of functional foods.

Development of Millettia speciosa champ polysaccharide conjugate stabilized oil-in-water emulsion for oral delivery of ß-carotene: Protection effect and in vitro digestion fate.

In this study, a natural antioxidant emulsifier, Millettia speciosa Champ polysaccharide conjugates (MSC-PC), was used for fabricating oil-in-water emulsion, and the influences of MSC-PC on ß-carotene stability and bioaccessibility were studied. Results suggested that MSC-PC stabilized emulsion exhibited excellent resistance to a wide range of salt levels (0-500 mM of Na+), thermal treatments (50-90 °C) and pH values (3.0-11.0). MSC-PC also exhibited an outstanding inhibition capacity on lipid oxidation. Besides, MSC-PC stabilized emulsion had a better protective effect on ß-carotene than other systems. Interestingly, in spite of similar lipolysis extent, ß-carotene bioaccessibility in MSC-PC fabricated emulsion (14.75 %) was markedly higher than that in commercial Tween 80 fabricated emulsion (10.08 %), likely due to the steric-hindrance effect and antioxidant ability of MSC-PC, building interfacial layers that prevented ß-carotene from degradation. This work supplied a deep insight into elucidating the mechanisms of emulsifying performance and ß-carotene protection effect of MSC-PC fabricated emulsion.

Flavonoid-enriched extract from Millettia speciosa Champ prevents obesity by regulating thermogenesis and lipid metabolism in high-fat diet-induced obese C57BL/6 mice.

Millettia speciosa (M. speciosa) Champ is a medicinal and edible plant. The roots are rich in flavonoids, which possess multiple biological activities, including lipid-lowering effects. This study aimed to explore the effect of flavonoid-enriched extract from M. speciosa (FMS) on obesity. The UPLC-Q-TOF-MS analysis and chromatographic analysis were adopted to identify flavonoid compounds in FMS. Male C57BL/6J mice were fed with a high-fat diet for 3 months and were then treated with FMS (50 or 100 mg/kg/d) or Orlistat (10 mg kg-1 d-1) for another 8 weeks. A total of 35 flavonoids were identified in the extract of M. speciosa root. FMS reduced body weight gain, liver weight gain, white adipose tissue, lipid accumulation, and blood glucose. The levels of TG, ALT, AST, and inflammatory-related adipokines (TNF-α and IL-6) in serum were also reduced by FMS. In addition, FMS promoted thermogenesis in brown adipose tissue and induced the activation of lipolysis, fatty acid oxidation, and oxidative phosphorylation in white adipose tissues. In summary, long-term administration of FMS could ameliorate high-fat diet-induced obesity by stimulating adipose thermogenesis and lipid metabolism.

Polysaccharides From the Roots of Millettia Speciosa Champ Modulate Gut Health and Ameliorate Cyclophosphamide-Induced Intestinal Injury and Immunosuppression.

Cyclophosphamide (CTX), a common anticancer drug, can cause a variety of side effects such as immunosuppression and intestinal mucosal injury. Polysaccharides are the major bioactive components of the roots of Millettia Speciosa Champ and have gained attention for their immunomodulatory activity. This study was designed to evaluate the immunomodulatory effect of Millettia Speciosa Champ polysaccharide (MSCP) on CTX-induced mice and the possible mechanism. The results showed that MSCP attenuated the CTX-induced decrease in body weight and immune organ indices in mice and promoted the secretion of immune-related cytokines (IL-2, IL-4, IL-10, TNF-α, and IgG). Meanwhile, MSCP restored intestinal morphology, increased the ratio of villus height/crypt depth (V/C), and improved the number of goblet cells and mucins expression. At the mRNA level, MSCP activated the TLRs/MyD88/NF-κB p65 pathway and enhanced the expression of genes related to intestinal mucosal integrity (Occludin1, Claudin1, and MUC-2). In addition, MSCP as a prebiotic improved microbial community diversity, regulated the relative abundance of dominant microbiota from the phylum level to the genus level, restored CTX-induced gut microbial dysbiosis, and promoted short-chain fatty acid production in mice. Based on the present findings, MSCP may modulate the immune response depending on enhancing intestinal health, suggesting that MSCP holds promise as a promising immunostimulant in functional foods and drugs.

Urinary metabonomics study of anti-depressive mechanisms of Millettia speciosa Champ on rats with chronic unpredictable mild stress-induced depression.

As a traditional Chinese medicine (TCM), Millettia speciosa Champ (MSC), exerts a wide range of pharmacological activities. Our research group previously found that MSC has antidepressant effects, but the specific antidepressant mechanisms remain unclear. Therefore, in this study, urine metabolomics based on ultra-performance liquid chromatography/quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) combined with pharmacodynamics was used to explore the pathogenesis of depression and the antidepressant effects of MSC. The results showed that MSC treatment could significantly improve chronic unpredictable mild stress (CUMS)-induced depression. Urine metabolic showed that the profiles of the CUMS model group were significantly separated from the control group, while the drug-treated groups were closer to the control group, especially the MSC group treated with a 14 g/kg dose of MSC. Furthermore, 9 metabolites, including glutaric acid, L-isoleucine, L-Dopa, sebacic acid, 3-methylhistidine, allantoin, caprylic acid, tryptophol, and 2-phenylethanol glucuronide, were identified as potential biomarkers of depression. Metabolic pathway analysis showed that these potential biomarkers were mainly involved in valine, leucine, and isoleucine biosynthesis, aminoacyl-tRNA biosynthesis, valine, leucine and isoleucine degradation, tyrosine metabolism, histidine metabolism, fatty acid biosynthesis, and pentose and glucuronate interconversions. Through Receiver operating characteristic (ROC) analysis and Pearson correlation analysis, the combination of L-isoleucine, sebacic acid, and allantoin, were further screened out as potential pharmacodynamic biomarkers associated with the efficacy of MSC. This study suggests that the integration of metabolomics with pharmacodynamics helps to further understand the pathogenesis of depression and provides novel insight into the efficacy of TCM.

Ameliorating effect on glycolipid metabolism and chemical profile of Millettia speciosa champ. extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Millettia speciosa Champ., also called Niu dali, is a fabaceous medicinal plant mainly distributed in southeast China, where it is a functional food for developing physical strength, and often used traditionally in medicinal treatment of numbness of the wrists, diabetes, hepatitis, and so on. AIM OF THE STUDY: To investigate the chemical profile, ameliorating effects of MSC on glycolipid metabolism in diabetic mice and to identify the possible mechanism of action. MATERIALS AND METHODS: High-performance liquid chromatography coupled with electrospray ionization quadrupole time of flight mass spectrometry (HPLC-ESI-QTOF-MS) was applied to analyze the chemical compositions from M. speciosa extract (MSC). MSC was orally administered to high-fat diet and STZ-induced diabetic mice at doses of 4.55, 9.10 and 13.65 mg/(kg·d) respectively for 10 weeks. Indices of glycolipid metabolism, including fasting blood glucose (FBG), fasting insulin, insulin resistance index (IRI), blood lipids, HPA-axis hormones, and related gene expressions were evaluated. RESULTS: 86 compounds were tentatively identified from MSC, counting for 91.97% of the total extract, mainly including 23 alkaloids (including 2 cyanogenetic glycosides firstly identified in this species, total content accounted to 39.71%), 23 flavonoids (11.91%), 17 acids (including 3 amino acids, 9 phenolic acids and 5 organic acids; 9.2%), 9 terpenoids and steroids (20.13%), 7 esters (3.33%), 3 lignans (3.73%), 3 saccharides (4.0%) and 1 anthraquinone (0.18%). MSC could ameliorate the glycolipid disorder in diabetic mice markedly, and significant regulations on CRH and ACTH hormones were observed. Moreover, the cellular morphology of liver and pancreas were significantly improved and the expressions of IRS2, PI3K, Akt and GLUT4 were significantly up-regulated by MSC treatment. CONCLUSION: This was the first time to study the chemical profile and ameliorating effect on glycolipid metabolism of M. speciosa. It was found to be rich in flavonoids and alkaloids, which might support the potential relation of material foundation and the activity in regulating glycolipid metabolism. The ameliorating effect on glycolipid disorder in diabetic mice might be associated to the regulation of related hormones of the HPA axis and the IRS2/PI3K/Akt/GLUT4 signalling pathway. It was of great significance for advanced directed separation and pharmacological activity research of MSC.

Metabolomic profiling of M. speciosa champ at different growth stages.

Millettia speciosa Champ (M. speciosa) is an edible food and folk medicine and extracts from its roots exhibit a hepatoprotective effect. However, its metabolic growth process and the best harvest time have not been reported. This study systematically evaluated the metabolomic profiling of M. speciosa root extracts at different growth stages through the UPLC-Q-TOF-MS, nuclear magnetic resonance (NMR) and An orthogonal partial least squares-discriminant analysis (OPLS-DA). The results revealed there were significant differences among extracts from six ages of M. speciosa, and 110 compounds were identified. Pharmacological studies showed that 5-year and 20-year old M. speciosa roots may exhibit higher fat-lowering effects, while 5-year-old (M.s-5Y) showed better hepatoprotective activity in high-fat diet (HFD)-induced nonalcoholic fatty liver disease (NAFLD) mice. Hence, our study suggested that M.s-5Y may have potent efficacy in ameliorating NAFLD, which might be useful in determining the optimum time to harvest M. speciosa.

A novel polysaccharide from the roots of Millettia Speciosa Champ: preparation, structural characterization and immunomodulatory activity.

A novel polysaccharide fraction (MSCP2) was extracted and isolated from the roots of Millettia Speciosa Champ. Structural characterization revealed that MSCP2 had an average molecular weight of 2.85 × 104 Da and was composed of fucose, arabinose, galactose, glucose and xylose with a ratio of 2.20: 2.52: 4.04: 87.29: 3.96. Methylation analysis and nuclear magnetic resonance (NMR) analysis showed that the main glycosidic linkage types of MSCP2 were proved to be α-D-Glcp-(1→, →4)-α-D-Glcp-(1→, →4)-α-D-Xylp-(1→, →6)-ß-D-Galp-(1→, α-L-Araf-(1→, →3,4)-ß-L-Fucp-(1→ and →4)-α-D-GalpA-(1→. The immunomodulatory assay suggested that MSCP2 could significantly improve the pinocytic capacity and increase the secretion of nitric oxide (NO) and cytokines by regulating the corresponding mRNA expression in RAW 264.7 cells. The data from the membrane receptor assay demonstrated that the potential mechanisms of MSCP2-induced macrophage activation were mainly through toll-like receptor 4 (TLR4), scavenger receptor type A (SRA) and glucan receptor (GR)-mediated signaling pathways. These results suggested that MSCP2 can be developed as a promising immunomodulatory agent in functional foods.

Determination of eight kinds of metal elements in millettia speciosa champ by icp-ms / 中国药学杂志

OBJECTIVE: To establish a method for simultaneous determination of multiple metal elements in Millettia speciosa Champ. METHODS: Eight kinds of metal elements, i.e., Fe, Mn, Cu, Zn, Pd, Cd, As, and Se in Millettia speciosa Champ from different origins were determined by microwave digestion and ICP-MS. RESULTS: The contents of Fe, Mn, Cu, Zn, Pd, Cd, As, and Se in Millettia speciosa Champ from different origins were slightly different, the contents of Fe, Mn, Cu, and Zn were all above 1 mg·kg-1, while the contents of Pd, Cd, As and Se were mostly less than 1 mg·kg-1. The linear correlation coefficient of each metal element was 0.999 2-1.000 0, the average recovery was 87%-110.8%, and the RSDs of the reproducibility test was less than 10%. CONCLUSION: A microwave digestion and ICP-MS method for simultaneous determination of multiple metal elements in Chinese medicine Millettia speciosa Champ is established. The method is simple, sensitive, precise and repeatable, which can be used to determine the contents of metals in Millettia speciosa Champ.

Effects of Polysaccharides from Millettia speciosa Champ on Proliferation of Spleen Lymphocyte and Secretion of Cytokine in Mice / 医药导报

Objective To explore the mechanism of immunoregulation by investigating the effects of polysaccharides from Millettia speciosa Champ (MSC) on proliferation of spleen lymphocyte and secretion of cytokine in mice.Methods The effects of MSC polysaccharides on Con A-induced spleen T lymphocyte proliferation were determined by MTT.The contents of TNF-α, IL-6 and PGE2 were determined by ELISA.Results The Con A-induced T lymphocyte proliferation was significantly increased by MSC polysaccharides at the concentrations from 50 to 200 μg·mL-1.Coupled with TNF-α and IL-6 were significantly increased while PGE2 was significantly decreased.Conclusion MSC polysaccharides could increase proliferation of spleen lymphocyte and enhance the immune responses by increasing the secretion of TNF-α and IL-6 in mice.

Chemical constituents from roots of Millettia speciosa / 中草药

Objective: To study the chemical constituents from the roots of Millettia speciosa (Leguminosae). Methods: Compounds in the 95% ethanol extract from the air-dried roots of M. speciosa were isolated by chromatography on silica gel column together with recrystallization, and their structures were identified by their physicochemical characteristics and spectral features. Results: Sixteen components were isolated from the air-dried roots of M. speciosa and identified as 7-oxo-β-sitosterol (1), aurantiamide acetate (2), shionone (3), maleic acid (4), psoralen (5), N-methylcytisine (6), lupeol caffeate (7), bisdemethoxycurcumin (8), vanillic acid (9), syringic acid (10), 6-methoxydihdyrosanguinarine (11), glycyrrhizic acid (12), (E)-3, 3'-dimethoxy-4, 4'-dihydroxystilbene (13), schisandrol B (14), 7-hydroxylathyrol (15), and nardosinone (16). Conclusion: All the compounds are isolated from this plant for the first time, and compounds 3, 7, 8, 11, and 13-16 are isolated from the plants of Leguminosae for the first time, compounds 2, 4-6, 9, 10 are isolated from the plants of Millettia Wight et Arn. for the first time.