Discovering the main "reinforce kidney to strengthening Yang" active components of salt Morinda officinalis based on the spectrum-effect relationship combined with chemometric methods.

Morinda officinalis, a well-known traditional herbal medicine in China, is used to treat deficiency of kidney-yang syndrome. Although this medicine has the property of "reinforcing kidney to strengthening Yang," the chemical constituents responsible for this effect remain to be elucidated. Here, we aimed to identify the main active compounds responsible for reinforcing kidney to strengthening Yang, based on spectrum-effect relationships combined with chemometrics. We used the UPLC-diode array detection method to establish the chromatography fingerprint of M. officinalis. Hydrocortisone-induced and adenine-induced kidney-yang deficiency patterns were established to evaluate the efficacy of M. officinalis. Serum triiodothyronine, free thyroxine, thyrotropin, testosterone, cortisol, luteinizing hormone, follicle-stimulating hormone, corticotropin-releasing hormone, and adrenocorticotropic hormone levels were determined as pharmacodynamic indices. Analytic hierarchy process was used to determine the weight of each index to the total pharmacodynamic contribution. Lastly, the spectrum-effect between the fingerprint and the pharmacological effects were established using grey relational analysis and partial least squares. Our findings indicated that peaks 1, 2, 3, 5, 6, 7, 8, 9, 11, 13, 15, 17, and 20 might represent the main components that positively correlated to the total effect, of which four were identified by comparison with reference standards. The identified components were monotropein (peak 1), deacetyl asperulosidic acid (peak 3), asperulosidic acid (peak 8), and asperuloside (peak 9). Our results suggest that the "reinforce kidney to strengthening Yang" effects were attributable to the combined effects of the multiple chemical components of M. officinalis and provide a valuable method to identify the active "reinforce kidney to strengthening Yang" components of M. officinalis and establish the quality control of M. officinalis.

Beta-1,6 glucan converts tumor-associated macrophages into an M1-like phenotype.

Tumor-associated macrophages (TAMs) with an M2-like phenotype have been linked to immunosuppression and resistance to chemotherapies of cancer, thus targeting TAMs has been an attractive therapeutic strategy to cancer immunotherapy. We have reported that the ß-D-(1→6) glucan (AAMP-A70) isolated from Amillariella Mellea could promote macrophage activation. The present study showed that the ß-1,6-glucan could promote the transformation of M2-like macrophages to M1-like phenotype and inhibit the viability of colon cancer cells in vitro and in vivo. On a cellular mechanistic level, the ß-1,6-glucan reset tumor-promoting M2-like macrophages to tumor-inhibiting M1-like phenotype via increasing the phosphorylation of Akt/NF-κB and MAPK. Further, TLR2 was identified as the receptor of ß-1,6-glucan in the transformation effect. In addition, a very similar ß-1,6-glucan with side chains of ß-Glc or α-Galρ which was purified from Lentinus edodes showed same activities with those from Amillariella Mellea. Our findings shed light on the action mode of ß-1,6-glucan in cancer immunotherapy.

Preparation, structural characterization, and bioactivity of PHPD-IV-4 derived from Porphyra haitanensis.

Polysaccharides are a major active component of Porphyra haitanensis, which is an important food source in many countries. Four different molecular-weight fractions, namely PHPD-I (329 kDa), PHPD-II (203 kDa), PHPD-III (128 kDa), and PHPD-IV (10 kDa), were obtained from P. haitanensis polysaccharides by degradation using the H2O2/ascorbic acid system. PHPD-IV elicited the highest level of antioxidant and immunostimulatory activity among the four fractions. PHPD-IV was purified by DEAE-cellulose column and five fractions were obtained, designated PHPD-IV-1-PHPD-IV-5. PHPD-IV-4 displayed the greatest biological activity by up-regulating the phosphorylation of MAPK signalling molecules. PHPD-IV-4 was further purified, and its structure was characterized by monosaccharide composition and 1/2D-NMR analysis. The result revealed that PHPD-IV-4 was repeated units of â†’ 3) ß-d-galactose (1 â†’ 4) 3, 6-anhydro-α-l-galactose (1→, and â†’ 3) ß-d-galactose (1 â†’ 4) α-l-galactose-6-S (1→. This study provides a theoretical basis for the utilisation and structure-activity assessment of P. haitanensis polysaccharides.

Spectrum-effect relationship between fingerprints and hemopoietic effects of small molecular fraction of Polygoni Multiflori radix praeparata.

Polygoni multiflori Radix Praeparata (PMRP) is a traditional medicine used for nourishing essence and blood in China. However, it is unclear which PMRP compounds are responsible for its hematopoietic effect. In this study, spectrum-effect relationship was used to discovery potential hematopoietic compounds. The fingerprints of 20 PMRP batches were established by HPLC and the hematopoietic effect was determined using red blood cell, hemoglobin, hematocrit, and platelet indexes in aplastic anemia model mice. The spectrum-effect relationship between common peaks and hematopoietic efficacy values was established using gray relational analysis and partial least squares analysis. Spectrum-effect relationship results showed that peaks 21 (emodin-8-O-(6´-O-acetyl)-ß-D-glucoside), 15 (2, 3, 5, 4'-tetrahydroxystilbene-2-O-di-glucoside), 16 (cis-2,3,5,4'-tetrahydroxy-stilbene-2-O-ß-D-glucoside), 11 (unknown), 20(unknown, 12 (epicatechin), 29 (carboxyl emodin), and 31 (emodin) in the fingerprints were closely related to the hematopoietic effect. This work successfully established the spectrum-effect relationship between PMRP hematopoietic effect and its fingerprints, which can be used to explain the material basis for the PMRP hematopoietic effect.

Purification, structural characterization, and immunomodulatory activity of the polysaccharides from Ganoderma lucidum.

A preliminary relationship was illustrated between the structural characteristics and corresponding immunomodulatory activities of G. lucidum polysaccharides. Two polysaccharides (GLP-1 and GLP-2) were purified from Ganoderma lucidum extracts by gradient ethanol precipitation and a Q-Sepharose Fast Flow (QFF) strong anion-exchange column. The monosaccharide composition, high-performance gel permeation chromatography-multi-angle laser light scattering-refractive index (HPGPC-MALLS-RI), Fourier-transform infrared spectroscopy (FT-IR), atomic force microscopy (AFM), methylation analysis, and nuclear magnetic resonance (NMR) were used to characterize these polysaccharides. The GLP-1 polysaccharide was elucidated as d-galactoglucan with a flexible random linear conformation that mainly composed of →6)-ß-d-Glcp-(1→, →6)-α-d-Galp-(1→, and →3)-ß-d-Glcp-(1→ residues. GLP-2 was found to be a relatively homogeneous ß-d-glucan that possessing →6)-ß-d-Glcp-(1→ and →3)-ß-d-Glcp-(1→ residues packaged into a spherical conformation. Immunomodulatory activities in vivo demonstrated that GLP-1 produced better protection of the spleen and thymus and was more effective for promoting hematopoiesis and improving IgA levels in serum. Our results suggest that the immunomodulatory activities of G. lucidum polysaccharides are highly corresponded to their structural characteristics such as carbohydrate composition, molecular weight and advanced conformation. This study provides a preliminary basis for studying the relationship between polysaccharide structure characterization and pharmacological activities.

Hematopoietic effect of small molecular fraction of Polygoni multiflori Radix Praeparata in cyclophosphamide-induced anemia mice.

The aim of this study is to investigate the protective effects of a small molecular fraction (SMF) of Polygoni multiflori Radix Praeparata (PMRP) in a cyclophosphamide (CTX) induced anemia mouse model. Small molecular fraction of PMRP was prepared and identified by high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). In pharmacology, we examined the peripheral hemogram and thymus and spleen index. The content of granulocyte-macrophage colony-stimulating factor (GM-CSF) in serum was mensurated by enzyme-linked immunosorbent assay (ELISA); The level of superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) in serum and spleen tissue homogenate were detected, and glutathione peroxidase (GSH-PX) was assayed in spleen. The results show that SMF can significantly accelerate the recovery of peripheral hemogram, increase the activity of antioxidant enzymes and GM-CSF in serum and spleen. SMF also increases the number of spleen cells, improves bone marrow pathology. In conclusion, the SMF of PMRP promoted the recovery of hematopoietic function in a CTX-induced anemia mouse, which can support SMF to be used as an adjunct to chemotherapy to counteract its side effects.

Alkaline Extraction, Structural Characterization, and Bioactivities of (1→6)-ß-d-Glucan from Lentinus edodes.

The purpose of this study is to develop a robust approach to obtain ß glucans from Lentinus edodes and to characterize their structural and biological properties for sustainable utilization. The alkali extraction was optimized with an orthogonal experimental design, and a concise process for obtaining specific targeting polysaccharides from Lentinus edodes was developed in this study. After purification with a Q-Sepharose Fast Flow strong anion-exchange column, the monosaccharide composition, a methylation analysis, and NMR spectroscopy were employed for their structural characterizations. LeP-N2 was found to be composed of (1→6)-ß-d-glucans with minor ß-(1→3) glucosidic side chains. Atomic force microscopy (AFM) and high-performance gel permeation chromatography-refractive index-multi-angle laser light scattering (HPGPC-RI-MALLS) also revealed LeP-N2 exhibiting a compact unit in aqueous solution. This (1→6)-ß-d-glucan was tested for antioxidant activities with IC50 at 157 µg/mL. Moreover, RAW 264.7 macrophage activation indicated that the release of nitric oxide (NO) and reactive oxygen species (ROS) was markedly increased with no cytotoxicity at a dose of 100 µg/mL. These findings suggest that the (1→6)-ß-d-glucans obtained from Lentinus edodes could serve as potential agents in the fields of functional foods or medicine.

Extraction, isolation and structural characterization of a novel polysaccharide from Cyclocarya paliurus.

A water soluble polysaccharide CP-III was extracted and purified from Cyclocarya paliurus. CP-III is identified as a novel pectin-like polysaccharide with molecular weight (Mw) of 72.7 kDa. The structural features of CP-III were characterized by methylation and nuclear magnetic resonance (NMR) spectroscopy. Its depolymerized fragments were analyzed by hydrophilic interaction chromatography-Fourier transform mass spectrometry (HILIC-FTMS). The main chain of CP-III is composed of →4)GalAp(α1 → and →2)Rhap(α1 → 4)GalAp(α1→, repeatedly. The residue of →4)Galp(ß1 → and →5)Araf(α1 → alternately exists on the O-4 of partial →2)Rhap(α1 → residues as side chains. On the O-3 of sectional →5)Araf(α1 → residues is a secondary branch assembled by →3)Araf(α1→. Moreover, on the non-reducing terminus of →4)Galp(ß1 → occasionally have an →5)Araf(α1 → chain. Surprisingly, a sub-branch constructed by →6)Hexp(ß1 → with a galacturonate or methyl galacturonate exists on the O-3 of certain →4)Galp(ß1 → residues in the non-reducing terminus. In addition, a terminal Xyl is located on the O-3 of fractional →4)GalAp(ß1 → residue. The highly branched polysaccharide CP-III with high water solubility can be used as food supplement and medicinal carrier in the future.

[Rapid identification of geographical origins and determination of polysaccharides contents in Ganoderma lucidum based on near infrared spectroscopy and chemometrics].

Near infrared spectroscopy combined with chemometrics methods was used to distinguish Ganoderma lucidum samples collected from different origins, and a prediction model was established for rapid determine polysaccharides contents in these samples. The classification accuracy for training dataset was 96.87%, while for independent dataset was 93.33%; as for the prediction model, 5-fold cross-validation was used to optimize the parameters, and different signal processing methods were also optimized to improve the prediction ability of the model. The best square of correlation coefficients for training dataset was 0.965 4, and 0.851 6 for validation dataset; while the root-mean-square deviation values for training dataset and validation dataset were 0.018 5 and 0.023 6, respectively. These results showed that combining near infrared spectroscopy with suitable chemometrics approaches could accuracy distinguish different origins of G. lucidum samples; the established prediction model could precious predict polysaccharides contents, the proposed method can help determine the activity compounds and quality evaluation of G. lucidum.