Polysaccharide from Angelica Sinensis Suppresses Inflammation and Reverses Anemia in Complete Freund's Adjuvant-induced Rats.

The anti-inflammatory and antianemic activities of Angelica sinensis polysaccharide (ASP) isolated from roots of Angelica sinensis (AS) was investigated in a complete Freund's adjuvant (CFA)-induced arthritic rat model. It was observed that serum iron (SI) and total iron binding capacity (TIBC) levels were elevated after 4-week oral administration of ASP. Red blood cell (RBC) count and hemoglobin (Hb) concentrations were ameliorated as well. Moreover, inflammatory cytokines IL-6 and TNF-a were decreased strikingly in CFA-induced arthritic rats after treatment of ASP. Evidence also showed that ASP strongly inhibited hepcidin expression through the Janus kinase/signal transducers and activators of transcription (JAK2/STAT3) pathway. Furthermore, ASP exhibited reduced primary and secondary lesions in adjuvant arthritis, attenuating synovitis and inflammatory joint damage. Data presented in this article collectively indicated that ASP significantly decreased proinflammatory cytokines (TNF-a, IL-6), which might play a crucial role in the CFA-induced arthritic rats, and had a therapeutic effect on adjuvant arthritis in rats. Results of Western blot analysis indicated that ASP inhibited the activation of IL-6/JAK2/STAT3 signaling pathway in the CFA-induced arthritic rats.

Biomimetic Matrix Stiffness Modulates Hepatocellular Carcinoma Malignant Phenotypes and Macrophage Polarization through Multiple Modes of Mechanical Feedbacks.

The extracellular matrix (ECM) stiffening is an important sign of local microenvironment change, which is considered as a hallmark of many diseases including hepatocellular carcinoma (HCC). The fates of both cancer cells and immune cells can be regulated by mechanical feedbacks acquired from ECM, but there is a lack of a precise study of mechanical feedback modes in different cell phenotypes following with the progressively increasing ECM stiffness. Herein, we used a biopolymeric film without further modification of adhesive molecules, as a natural local niche to mimic a gradually stiffening manner from HCC onset in liver cirrhosis to its metastasis in the spinal cord. Three distinct manners of mechanical feedbacks were found: the gradual manner in HCC cell spreading, migration and early apoptosis to oxaliplatin, the stepwise manner in HCC cell adhesion, proliferation, focal adhesion (FA) formation, drug resistance, and macrophage M1 polarization; the specific manner in the stages of the progression of epithelial-mesenchymal transition at different stiffness ranges. Further investigation of molecular mechanisms confirmed those mechanical feedback manners by signaling activation of FA kinase, phosphatidylinositol 3-kinase, and expression of pro-/antiapoptotic and pro-/anti-inflammatory genes. Our results pave a novel avenue to know about mechanical feedbacks from ECM, which could be used for future cancer studies and in vitro drug screening applications.

Immobilized Transforming Growth Factor-Beta 1 in a Stiffness-Tunable Artificial Extracellular Matrix Enhances Mechanotransduction in the Epithelial Mesenchymal Transition of Hepatocellular Carcinoma.

Cancer progression is regulated by multiple factors of extracellular matrix (ECM). Understanding how cancer cells integrate multiple signaling pathways to achieve specific behaviors remains a challenge because of the lack of appropriate models to copresent and modulate ECM properties. Here we proposed a strategy to build a thin biomaterial matrix by poly(l-lysine) and hyaluronan as an artificial stiffness-tunable ECM. Transforming growth factor-beta 1 (TGF-ß1) was used as a biochemical cue to present in an immobilized and spatially controlled manner, with a high loading efficiency of 90%. Either soft matrix with immobilized TGF-ß1 (i-TGF) or bare stiff matrix could only promote HCC cells to form the epithelial phenotype, whereas stiff matrix with i-TGF was the only condition to induce the mesenchymal phenotype. Further investigation revealed that i-TGF increased the specific TGF-ß1 receptor (TßRI) expression to activate PI3K pathway. i-TGF-TßRI interactions also promoted HCC cell adhesion to enlarge contact area for stiffness sensing, resulting in the raising expression of the mechano-sensor (ß1 integrin). Mechanotransduction would then be enhanced by the ß1 integrin/vinculin/p-FAK pathway, leading to a noble PI3K activation. Using our model, a novel mechanism was discovered to elucidate regulation of cell fates by coupling mechanotransduction and biochemical signaling.

Phosphorus-doped graphene-based electrochemical sensor for sensitive detection of acetaminophen.

Phosphorus-doped graphene (P-RGO) was synthesized and employed as active electrode material to construct electrochemical sensor for acetaminophen (AP). The P-RGO coated glass carbon electrode (P-RGO/GCE) showed an excellent electrocatalytic activity for the oxidation of AP, resulted from highly enhanced electrochemical conductivity and accelerated electron transfer. The experimental conditions for AP detection were optimized, and under the optimal condition, a linear relationship between current intensity and concentration of AP was obtained in the range of 1.5-120 µM with a detection limit of 0.36 µM (S/N = 3). The developed sensor showed high selectivity for AP in the presence of various common species, excellent reproducibility and stability. The present sensor was also successfully applied for AP detection in pharmaceutical tablet samples.

Pharmacokinetics, biodistribution and receptor mediated endocytosis of a natural Angelica sinensis polysaccharide.

The interest in developing new drug carriers for delivery to the liver using natural polysaccharides with a high galactose content has necessitated the study of the pharmacokinetics and tissue distribution of these polysaccharides. In this paper, a new method was established for the microanalysis of Angelica sinensis polysaccharide (ASP) in biosamples. Fluorescein-labelled ASP (FA) was rapidly eliminated from the bloodstream and distributed to the liver with high specificity following intravenous injection. The analysis of the hepatocellular localization demonstrated that FA was predominantly endocytosed by the parenchymal cells, an observation consistent with the results obtained from microscopy studies. Additionally, FA showed a high affinity for asialoglycoprotein receptor-rich cells, while minimal binding of FA to asialoglycoprotein receptor-poor cells was observed. Moreover, the absorption of FA was markedly inhibited by the co-administration of neogalactosylalbumin (NGA) both in vivo and in vitro. To allow for the visualization of the systemic circulation of ASP, 99mTc-DTPA-ASP was synthesized and in vivo imaging was performed with single photon emission computed tomography (SPECT). It also showed a high aggregation of 99mTc-DTPA-ASP in liver. These results suggest that the distribution of ASP to the liver occurs via asialoglycoprotein receptor (ASGPR) mediated endocytosis and ASP could potentially be applied as a new carrier for delivering drugs to the liver.

Structural characterization and in vitro antitumor activity of an acidic polysaccharide from Angelica sinensis (Oliv.) Diels.

A water-soluble polysaccharide extracted from the roots of Angelica sinensis (Oliv.) Diels, which is a traditional Chinese medicine herb, was fractioned and purified by Sephadex G-50 gel filtration chromatography. The structural characterization and antitumor activities of the purified polysaccharide fraction, named as ASP, were evaluated in the present study. ASP, which molecular weight was determined to be 80kDa by high-performance gel-permeation chromatography, is an acidic heteropolysaccharide consisting of glucuronic acid, glucose, arabinose and galactose in ratio of 1.00:1.70:1.85:5.02. It has a backbone composed of (1→3)-linked Galp, (1→6)-linked Galp and 2-OMe-(1→6)-linked Galp with three branches attached to O-3 of 2-OMe-(1→6)-linked Galp and terminated with GlcpA and Araf, and all of Araf and the majority of Glcp are distributed in branches. Moreover, all of GlcpA were presented as (1→)-linked GlcpA in branches. In in vitro antitumor assays, ASP displayed cytotoxicity against HepG2 cells (34.32±3.50% at the concentration of 1mg/mL) and MCF-7 cells (28.90±1.50% at the concentration of 1mg/mL) in a dose-dependent manner, and ASP also showed mild inhibitory activity against A549 cells.

Alkali-soluble polysaccharide, isolated from Lentinus edodes, induces apoptosis and G2/M cell cycle arrest in H22 cells through microtubule depolymerization.

The aim of the study was to evaluate the pro-apoptotic effects of polysaccharides derived from Lentinus edodes and further elucidated the mechanisms of this action. Our results demonstrated that marked morphological changes of apoptosis were observed after treatment of L. edodes polysaccharides [Lentinan (LTN)]. Moreover, LTN-induced cell apoptosis was characterized by a rapid stimulation of reactive oxygen species production, the loss of mitochondrial membrane potential and an increase in intracellular concentration of Ca(2+) . In addition, the results of the haematoxylin and eosin and TUNEL assay further confirmed that LTN-induced apoptosis in vivo. Furthermore, flow cytometry analysis showed that LTN could arrest the cell cycle at G2/M phase, and immunofluorescence showed LTN caused disruption of microtubule. These results suggest that disruption of cellular microtubule network, arrest of the cell cycle at G2/M phase and induction of apoptosis may be one of the possible mechanisms of anti-tumour effect of LTN.

Structure and inducing tumor cell apoptosis activity of polysaccharides isolated from Lentinus edodes.

In this study, five novel polysaccharides SLNT1, SLNT2, JLNT1, JLNT2, and JLNT3 were isolated from the fruit body of Lentinus edodes. Chemical and physical analyses showed that the five polysaccharides consist of glucose with the structure of ß-(1→3)-d-glucose main chains and ß-(1→6)-d-glucose side chains. Moreover, all of them had triple-helical conformation and different molecular weight distributions. Animal studies further demonstrated that the antitumor effects were remarkably improved by SLNT1 and JLNT1 treatments with the inhibitory rates of 65.41% and 61.07% in H22-bearing mice, respectively. Additionally, both of them significantly increased the levels of serum IL-2 and TNF-α production and induced the tumor cell apoptosis. Taken together, our findings revealed that the involved antitumor mechanisms possibly in part were mediated not only by enhancing the immunity but also by directly killing the tumor and the induction of tumor cell apoptosis in H22-bearing mice.

Study to establish the role of JAK2 and SMAD1/5/8 pathways in the inhibition of hepcidin by polysaccharides from Angelica sinensis.

ETHNOPHARMACOLOGICAL RELEVANCE: Angelica sinensis polysaccharide (ASP) is one of the major active ingredients in Angelica sinensis (Oliv.) Diels. This traditional Chinese medicine has been used for thousands of years for treating gynecological diseases. AIM OF THE STUDY: Previous studies have suggested that ASP from the roots of Angelica sinensis (Oliv.) Diels suppresses hepcidin expression, but the underlying molecular mechanisms are not known. The present study was designed to establish the role of the janus-kinases 2 (JAK2) and son of mothers against decapentaplegic 1/5/8 (SMAD1/5/8) pathways in the inhibition of hepcidin by polysaccharides from Angelica sinensis in normal rats. MATERIALS AND METHODS: ASP was administered orally (0.3, 0.6 and 1.2g/kg body weight) to male Sprague-Dawley rats every day for 20 days. Intraperitoneal injections of recombinant human erythropoietin (rhEPO; 800 and 2000U/kg body weight) were given to the positive control group every day for 3 days. After administration, hepcidin levels, blood parameters, serum iron status and non-heme iron concentrations in the liver were examined. Western blot analyses were used to investigate the expression of five relevant signaling proteins in the liver. RESULTS: RhEPO injection significantly stimulated erythropoiesis and expression of the serum transferrin receptor (sTfR), and decreased serum iron status and non-heme iron concentrations in the liver. However, blood parameters barely changed in the ASP groups. sTfR, serum iron, and liver iron levels altered only in the ASP high-dose group (1.2g/kg body weight). rhEPO and ASP significantly reduced hepcidin expression by inhibiting the expression of phospho-SMAD1/5/8 and JAK2 in the liver, but not through transmembrane protease serine 6 (TMPRSS6) and extracellular signal-regulated kinase 1/2 (ERK1/2). CONCLUSIONS: These data suggested that ASP can interrupt the JAK2 and SMAD1/5/8 pathways, which eventually results in lower expression of hepcidin.

Polysaccharide isolated from Angelica sinensis inhibits hepcidin expression in rats with iron deficiency anemia.

A novel polysaccharide named Angelica sinensis polysaccharide (ASP) was obtained from the powdered and defatted roots of A. sinensis (Oliv.) Diels. The molecular weight of ASP was determined to be 78 kDa and was 95.0% sugars consisting of mostly arabinose, glucose, and galactose with a molar ratio of 1:5.68:3.91. A previous study indicated that ASP may increase plasma iron levels by suppressing the expression of hepcidin, a negative regulator of body iron metabolism, in the liver. The present study aims to clarify the inhibitory effect of ASP on hepcidin expression in rat models of iron deficiency anemia (IDA), and clarify the mechanisms involved. It was demonstrated that ASP significantly reduced hepcidin expression by inhibiting the expression of mothers against decapentaplegic protein 4 (SMAD4) in liver and stimulating the secretion of erythropoietin, which further downregulated hepcidin by repressing CCAAT/enhancer-binding protein α (C/EBPα) and the phosphorylation of signal transducer and activator of transcription 3/5. The results indicate that ASP can suppress the expression of hepcidin in rats with IDA, and may be useful for the treatment of IDA.

Inhibitory effect of polysaccharides isolated from Angelica sinensis on hepcidin expression.

ETHNOPHARMACOLOGICAL RELEVANCE: Angelica sinensis polysaccharide is an important bioactive component of Angelica sinensis (Oliv.) Diels that has been used in traditional Chinese medicine for treating gynecological disorders and anemia. AIM OF THE STUDY: Previous study indicated that Angelica sinensis polysaccharide (ASP) may promote plasma iron levels by suppressing the expression of hepcidin, a negative regulator of body iron metabolism, in the liver. The present study aims to clarify the inhibitory effect of ASP on hepcidin expression as well as the involved mechanisms. MATERIALS AND METHODS: ASP (1g/kg) or vehicle (normal saline) was intragastrically administrated to rats everyday for 14 d. Intraperitoneal injections of recombinant human erythropoietin (rhEPO, 2000 U/kg) were given to positive control group. Erythropoietin and hepcidin levels in serum at different time points were determined by enzyme-linked immunosorbent assay. Western blot was used to investigate the expression of 6 pertinent signal proteins in liver. RESULTS: ASP significantly reduced hepcidin expression by inhibiting the expression of signal transducer and activator of transcription 3/5 (STAT3/5) and mothers against decapentaplegic protein 4 (SMAD4) in liver and stimulating the secretion of erythropoietin, which further down-regulated hepcidin by repressing CCAAT/enhancer-binding protein α (C/EBPα), SMAD4, and the phosphorylation process of STAT3/5. CONCLUSIONS: ASP can suppress the expression of hepcidin in normal rats, and may be used in the treatments of hepcidin-induced diseases.

Zinc-doping in TiO2 films to enhance electron transport in dye-sensitized solar cells under low-intensity illumination.

A nanocrystalline TiO(2) film with highly dispersed Zn-doping shows its capability for efficient electron transport in dye-sensitized solar cells (DSSCs). The Zn-doping is conducted via Zn(2+) introduction into a layered titanate followed by hydrothermal treatment and calcination. The Zn-doped films exhibit an elevated electron Fermi level, which may enhance band bending to lower the density of empty trap states. Because of this Zn-doping, the consequent DSSCs can alleviate the decay of light-to-electric energy conversion efficiency due to light intensity reduction. Intensity-modulated spectroscopic analysis reveals that enhanced transport of photogenerated electrons as a result of the trap density minimization is responsible for the high cell performance under low-intensity illumination. A Zn-doping content of ca. 0.4 at% Zn/Ti can enhance the light conversion efficiency by 23% at a solar light intensity as low as 11 mW cm(-2). This technique can significantly extend the indoor application of DSSCs.

Effect of Angelica sinensis polysaccharide-iron complex on iron deficiency anemia in rats.

OBJECTIVE: To investigate the therapeutic effects of Angelica sinensis polysaccharide-iron complex (APIC) on rats with iron deficiency anemia (IDA). METHODS: The IDA rat model was established by adopting low-iron forage with a small amount of regular bloodletting. The rats were randomly divided into a model group, three AIPC groups (high, middle, and low dosage), an Angelica sinensis polysaccharide (ASP) group, a mixture group (ASP+FeCl(3)) and a positive control group (Niferex). Changes in hemoglobin (Hb), red blood cell count (RBC), hematocrit (HCT) and iron content of whole blood were observed. RESULTS: There was a significant difference before and after administration in all treated groups and all indices were restored to near-normal levels in the APIC groups and the positive control group. There was a significant difference among the changes of the indices in all the APIC groups and those of the model group but not between those of the APIC groups and the positive control group. However, the recovery of the indices in the APIC groups was superior to that in the positive control group. CONCLUSION: APIC not only has a superior therapeutic effect on IDA, but also has the effect of the ASP on supplementing blood and activating blood circulation. Hence, it may be used as a new iron-supplementing agent with a double therapeutic efficacy on blood supplementation for the treatment of IDA.

[Preliminary studies on physicochemical properties of Angelica sinensis polysaccharide-iron complex].

OBJECTIVE: To study some physicochemical properties of Angelica sinensis polysaccharide-iron complex (APC). METHOD: Based on the qualitatively identified reactions of iron (III), the qualitatively identified reactions of APC were found out by comparing hydroxide. The content of iron (III) in APC was determined with iodometry. The stability of APC under physiological pH conditions was judged by titrating APC with sodium hydroxide. The deoxidization of APC was tested with colorimetric analysis. RESULT: APC showed the qualitatively identified reactions of iron (III). The content of iron (III) in APC ranged from 10% to 40%, and the water-solubility of APC was related to the content of iron (III). The complex was stable at physiological pH from 3 to 12, without precipitation and dissociation. At 37 degrees C, Fe (III) in the complex was completely reduced to Fe (III) by ascorbic acid in about 6 hours. CONCLUSION: APC can be qualitatively identified by using the qualitatively identified reactions of iron (III). When its iron (III) content is within 20%-25%, APC has a better ability to dissolve in water. And the complex is stable under physiological pH conditions.