Investigation of the underlying mechanism of Buyang Huanwu decoction in ischemic stroke by integrating systems pharmacology-proteomics and in vivo experiments.

Buyang Huanwu Decoction (BHD) has been effective in treating ischemic stroke (IS). However, its mechanism of action remains unclear. The study intended to explore the potential mechanism of BHD against IS using systems pharmacology, proteomics, and animal experiments. The active components of BHD were identified from UPLC-Q-TOF-MS and literature mining. Systems pharmacology and proteomics were employed to investigate the underlying mechanism of BHD against IS. The AutoDock tool was used for molecular docking. A middle cerebral artery occlusion (MCAO) model rat was utilized to explore the therapeutic benefits of BHD. The rats were divided into sham, model, BHD (5, 10, 20 g/kg, ig) groups. The neurological scores, pathological section characteristics, brain infarct volumes, inflammatory cytokines, and signaling pathways were investigated in vivo experiments. The results of systems pharmacology showed that 13 active compounds and 112 common targets were screened in BHD. The docking results suggested that the active compounds in BHD had a high affinity for the key targets. In vivo experiments demonstrated that BHD exhibited neuroprotective benefits by lowering the neurological score, the volume of the cerebral infarct, the release of inflammatory cytokines, and reducing neuroinflammatory damage in MCAO rats. Furthermore, BHD decreased TNF-α and CD38 levels while increasing ATP2B2, PDE1A, CaMK4, p-PI3K, and p-AKT. Combined with systems pharmacology and proteomic studies, we confirmed that PI3K-Akt and calcium signaling pathways are the key mechanisms for BHD against IS. Furthermore, this study demonstrated the feasibility of combining proteomics with systems pharmacology to study the mechanism of herbal medicine.

Electrically-assisted anaerobic digestion under ammonia stress: Facilitating propionate oxidation and activating methanogenesis via direct interspecies electron transfer.

Electrical assistance is an effective strategy for promoting anaerobic digestion (AD) under ammonia stress. However, the underlying mechanism of electrical assistance affecting AD is insufficiently understood. Here, electrical assistance to AD under 5 g N/L ammonia stress was provided, by employing a 0.6 V voltage to the carbon electrodes. The results demonstrated remarkable enhancements in methane production (104.6 %) and the maximal methane production rate (207.7 %). The critical segment facilitated by electro-stimulation was the microbial metabolism of propionate-to-methane, rather than ammonia removal. Proteins in extracellular polymer substances were enriched, boosting microbial resilience to ammonia intrusion. Concurrently, the promoted humic/fulvic-substances amplified the microbial electron transfer capacity. Metagenomics analysis identified the upsurge of propionate oxidation at the anode (by e.g. unclassified_c__Bacteroidia), and the stimulations of acetoclastic and direct interspecies electron transfer-dependent CO2-reducing methanogenesis at the cathode (by e.g. Methanothrix). This study provides novel insights into the effect of electrical assistance on ammonia-stressed AD.

Precipitation pattern changed the content of non-structural carbohydrates components in different organs of Artemisia ordosica.

BACKGROUND: Non-structural carbohydrates (NSC) play a significant role in plant growth and defense and are an important component of carbon cycling in desert ecosystems. However, regarding global change scenarios, it remains unclear how NSCs in desert plants respond to changing precipitation patterns. [Methods] Three precipitation levels (natural precipitation, a 30% reduction in precipitation, and a 30% increase in precipitation) and two precipitation intervals levels (5 and 15 d) were simulated to study NSC (soluble sugar and starch) responses in the dominant shrub Artemisia ordosica. RESULTS: Precipitation level and interval interact to affect the NSC (both soluble sugar and starch components) content of A. ordosica. The effect of precipitation on NSC content and its components depended on extended precipitation interval. With lower precipitation and extended interval, soluble sugar content in roots increased and starch content decreased, indicating that A. ordosica adapts to external environmental changes by hydrolyzing root starch into soluble sugars. At 5 d interval, lower precipitation increased the NSC content of stems and especially roots. CONCLUSIONS: A. ordosica follows the "preferential allocation principle" to preferentially transport NSC to growing organs, which is an adaptive strategy to maintain a healthy physiological metabolism under drought conditions. The findings help understand the adaptation and survival mechanisms of desert vegetation under the changing precipitation patterns and are important in exploring the impact of carbon cycling in desert systems under global environmental change.

Soluble dietary fiber and cellulose from Saccharina japonica by-product ameliorate Loperamide-induced constipation via modulating enteric neurotransmitters, short-chain fatty acids and gut microbiota.

The effects of soluble dietary fiber (SDF) and cellulose (IDF) from Saccharina japonica by-product and their differences in improving constipation were further clarified in the present study. We demonstrated that SDF was mainly made up of d-mannuronic acid and d-mannose while IDF consisted of d-glucose , which is different from other reported dietary fibers of terrestrial plants. In this research, both SDF and IDF improved fecal-related indicators, gastrointestinal transit rate and histological morphology in Lop-induced mice. Moreover, they could increase the level of antioxidant enzymes (SOD and GSH-Px), restore the expression of enteric neurotransmitters, and maintain the function of ZO-1, JAM-1 as well as Occludin. Interestingly, SDF and IDF had a significant up-regulated effect on the proportion of Muribaculacea, Prevotellaceaen and Lachnospiraceae, which are critical to preserving intestinal immune homeostasis. Besides, they promoted the biosynthesis of short-chain fatty acids (SCFAs). The overall index showed that SDF is more effective for constipation due to its better water retention capacity. Thus, they can be used as a safe dietary supplement for the treatment of chronic or occasional constipation in humans.

Polysaccharide-Peptide from Trametes versicolor: The Potential Medicine for Colorectal Cancer Treatment.

The incidence and mortality of colorectal cancer have shown an upward trend in the past decade. Therefore, the prevention, diagnosis, and treatment of colorectal cancer still need our continuous attention. Finding compounds with strong anticancer activity and low toxicity is a good strategy for colorectal cancer (CRC) therapy. Trametes versicolor is a traditional Chinese medicinal mushroom with a long history of being used to regulate immunity and prevent cancer. Its extractions were demonstrated with strong cell growth inhibitory activity on human colorectal tumor cells, while the anticancer activity of them is not acted through a direct cytotoxic effect. However, the intricacy and high molecular weight make mechanistic research difficult, which restricts their further application as a medication in clinical cancer treatment. Recent research has discovered a small molecule polysaccharide peptide derived from Trametes versicolor that has a distinct structure after decades of Trametes versicolor investigation. Uncertain molecular weight and a complex composition are problems that have been solved through studies on its structure, and it was demonstrated to have strong anti-proliferation activity on colorectal cancer in vitro and in vivo via interaction with EGFR signaling pathway. It opens up new horizons for research in this field, and these low molecular weight polysaccharide peptides provide a new insight of regulation of colorectal cancer proliferation and have great potential as drugs in the treatment of colorectal cancer.

Conjugated linoleic acid ameliorates hepatic steatosis by modulating intestinal permeability and gut microbiota in ob/ob mice.

Background: Conjugated linoleic acid (CLA) is an effective supplement for reducing fat mass, but its effect on hepatic steatosis remains controversial. Objective: This study aims to evaluate the effect of CLA on liver fat accumulation, inflammation, gut microbiome, and intestinal barrier integrity. Design: Wild-type (WT) mice and ob/ob (OB) mice were randomly divided into four groups according to the treatment with/without 1% CLA: WT, WT mice treated with CLA (WT-CLA), OB, and OB mice treated with CLA (OB-CLA). Lipid metabolism and hepatic fat accumulation were evaluated by changes in histological and biochemical parameters. Gene expressions related to liver inflammation and intestinal barrier integrity were examined. The effect of CLA on the gut microbiota population was investigated. Results: The body weight, fatty tissue mass, and serum lipid levels of the WT-CLA group and OB-CLA group were separately lower than those of the WT group and OB group, but the livers of the WT-CLA group had more fatty lipids, higher triglyceride properties, and saturated fatty acid (FA) composition than those of the WT group, which was contrary to the effect of CLA on OB mice. Real time quantitative PCR results showed that CLA increased hepatic inflammation and intestinal permeability in the WT mice, while it significantly decreased the mRNA expression of liver TNF-α, IFN-γ, and IL-1ß and markedly ameliorated intestinal tight junction proteins in the OB mice. The gut microbiota testing indicated a higher abundance of beneficial bacteria (e.g., Lachnoclostridium, Roseburia, Dubosiella, Oscillibacter, and Anaerostipes) and a lower abundance of pro-inflammatory bacteria (e.g., Tyzzerella and Alistipes) in the OB-CLA group than those of the OB group. Correlation analysis suggested that gut microbiota correlated with liver inflammation, intestinal permeability, and hepatic FA composition. Conclusion: CLA potentially contributed to ameliorating hepatic steatosis in OB mice via modulating liver inflammation, intestinal permeability, and gut microbiota, which suggests CLA is more suitable for people with obesity or overweight.

Milk Exosomes Transfer Oligosaccharides into Macrophages to Modulate Immunity and Attenuate Adherent-Invasive E. coli (AIEC) Infection.

Exosomes are abundance in human body fluids like urine, milk and blood. They act a critical role in extracellular and intracellular communication, intracellular trafficking and physiological regulation. Multiple immune-modulatory components, such as proteins, RNAs and carbohydrates (glycoproteins), have been found in human milk exosomes, which play immune-regulatory functions. However, little is known about oligosaccharides in milk exosomes, the "free sugars", which act critical roles in the development of infant's immature mucosal immune system. In this study, the profile of milk exosomes encapsulated human milk oligosaccharides (HMOs) was calibrated with characteristic oligosaccharides in colostrum and mature milk, respectively. The exosomes containing human milk oligosaccharides were uptaken by macrophages, which were responsible for the establishment of intestinal immunity. Furthermore, mice pretreated with exosome encapsulated HMOs were protected from AIEC infection and had significantly less LPS-induced inflammation and intestinal damage. Exosome encapsulated milk oligosaccharides are regarded to provide a natural manner for milk oligosaccharides to accomplish their critical functions in modifying newborn innate immunity. The understanding of the interaction between a mother's breastfeeding and the development of an infant's mucosal immune system would be advantageous. The transport of milk oligosaccharides to its target via exosome-like particles appears to be promising.

Identification of Active Ingredients of Huangqi Guizhi Wuwu Decoction for Promoting Nerve Function Recovery After Ischemic Stroke Using HT22 Live-Cell-Based Affinity Chromatography Combined with HPLC-MS/MS.

OBJECTIVE: The Chinese medicine Huangqi Guizhi Wuwu Decoction (HGWD) has been reported to improve the clinical symptoms and restore nerve function after ischemic stroke; however, its active ingredients are not well-determined. Therefore, this study aimed to investigate the bioactive compounds of HGWD and explore the possible mechanism of action. METHODS: The methods, including live HT22 cells, solid-phase extraction, and HPLC-MS/MS were utilized. The potential ingredients were identified through comparisons with literature and monomer compounds. Then, oxygen-glucose deprivation reperfusion (OGD/R)-treated HT22 cells were utilized to investigate the effect of HGWD components with specific binding affinities. Reactive oxygen species (ROS), superoxide dismutase (SOD), lactate dehydrogenase (LDH), and Tunel staining were used as testing indexes to analyze the protective effects of potential active ingredients on OGD/R-induced damage. RESULTS: Eleven compounds with specific binding affinities were identified as calycosin-7-O-glucoside, calycosin, formononetin, cinnamic alcohol, cinnamic acid, betaine, dl-2-phenylpropionic acid, 4-hydroxycinnamic acid, 6-methylcoumarin, wogonin, and paeoniflorin. Among them, six compounds had a protective effect on OGD/R-treated HT22 cells. Furthermore, calycosin-7-O-glucoside, calycosin, paeoniflorin, 4-hydroxycinnamic acid, wogonin, and formononetin could regulate oxidative stress and apoptosis to attenuate the cell damage caused by OGD/R. CONCLUSION: The mechanism of action of HGWD to promote neurological recovery after ischemic stroke was related to the regulation of oxidative stress and apoptosis. This study suggested that cell membrane affinity chromatography combined with HPLC-MS/MS could be applied to screen potential active components in traditional Chinese medicines (TCM).

Effects of Dietary Supplementation with κ-Selenocarrageenan on the Selenium Accumulation and Intestinal Microbiota of the Sea Cucumbers Apostichopus japonicus.

A 30-day feeding trial was conducted to investigate the effect of κ-selenocarrageenan on the growth performance, selenium accumulation, antioxidant capacity, and intestinal microbiota of sea cucumbers Apostichopus japonicus, with different sizes (70 g ± 10 g and 100 g ± 10 g). Sea cucumbers of each size were randomly assigned into two groups; a diet without supplemented κ-selenocarrageenan was referred to as a control diet, or supplemented with κ-selenocarrageenan at selenium (Se) levels of 2.0 µg/g. Selenium accumulation in the body wall and intestine was determined on days 0, 10, 20, and 30. The survival rate (SR) was significantly higher in the κ-selenocarrageenan-treated group (Se group) than in the control group. After 30 days of feeding, κ-selenocarrageenan supplementation increased the activities of glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC), and decreased malondialdehyde (MDA) levels in A. japonicus. Furthermore, the intestinal microbiota diversity of sea cucumbers was increased by dietary supplementation with κ-selenocarrageenan and the relative abundances of some probiotics (such as Sulfitobacter and Rhodobacteraceae) were also increased. It is suggested that κ-selenocarrageenan could increase the antioxidant capacity and modulate the intestinal microbiota of sea cucumbers A. japonicus. Further researches will be conducted for its optimal administration concentrations in vivo.

Tanshinone IIA attenuates neuroinflammation via inhibiting RAGE/NF-κB signaling pathway in vivo and in vitro.

BACKGROUND: Glial activation and neuroinflammation play a crucial role in the pathogenesis and development of Alzheimer's disease (AD). The receptor for advanced glycation end products (RAGE)-mediated signaling pathway is related to amyloid beta (Aß)-induced neuroinflammation. This study aimed to investigate the neuroprotective effects of tanshinone IIA (tan IIA), a natural product isolated from traditional Chinese herbal Salvia miltiorrhiza Bunge, against Aß-induced neuroinflammation, cognitive impairment, and neurotoxicity as well as the underlying mechanisms in vivo and in vitro. METHODS: Open-field test, Y-maze test, and Morris water maze test were conducted to assess the cognitive function in APP/PS1 mice. Immunohistochemistry, immunofluorescence, thioflavin S (Th-S) staining, enzyme-linked immunosorbent assay (ELISA), real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR), and western blotting were performed to explore Aß deposition, synaptic and neuronal loss, microglial and astrocytic activation, RAGE-dependent signaling, and the production of pro-inflammatory cytokines in APP/PS1 mice and cultured BV2 and U87 cells. RESULTS: Tan IIA treatment prevented spatial learning and memory deficits in APP/PS1 mice. Additionally, tan IIA attenuated Aß accumulation, synapse-associated proteins (Syn and PSD-95) and neuronal loss, as well as peri-plaque microgliosis and astrocytosis in the cortex and hippocampus of APP/PS1 mice. Furthermore, tan IIA significantly suppressed RAGE/nuclear factor-κB (NF-κB) signaling pathway and the production of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1ß) in APP/PS1 mice and cultured BV2 and U87 cells. CONCLUSIONS: Taken together, the present results indicated that tan IIA improves cognitive decline and neuroinflammation partly via inhibiting RAGE/NF-κB signaling pathway in vivo and in vitro. Thus, tan IIA might be a promising therapeutic drug for halting and preventing AD progression.

Pharmacokinetic Analysis of Huangqi Guizhi Wuwu Decoction on Blood and Brain Tissue in Rats with Normal and Cerebral Ischemia-Reperfusion Injury by Microdialysis with HPLC-MS/MS.

OBJECTIVE: The aim of our research was to analyze and compare the pharmacokinetics of paeoniflorin, calycosin, calycosin-7-o-ß-d-6-glucoside, and 6-gingerol in the blood and brain tissue of normal and cerebral ischemia-reperfusion injury rats by HPLC-MS/MS method. METHODS: The blood and brain tissue samples of normal and middle cerebral artery occlusion (MCAO) rats were compared. The blood and brain tissue samples were collected by using microdialysis technique. The concentrations of paeoniflorin, calycosin, calycosin-7-o-ß-d-6-glucoside, and 6-gingerol in blood and brain tissues were determined by the HPLC-MS/MS internal standard method. RESULTS: Compared with the normal group, the model group after the administration of the Huangqi Guizhi Wuwu Decoction showed that Cmax blood, AUC0-t blood, and AUC0-inf blood of paeoniflorin were increased, CLblood, t1/2 brain, and Vbrain of paeoniflorin were decreased; Cmaxblood, AUC0-tblood, AUC0-infblood, and average residence time (MRTbrain) of calycosin-7-o-ß-d-6-glucoside were decreased and the CLblood and Cmax brain of calycosin-7-o-ß-d-6-glucoside were increased; Cmax blood of calycosin was decreased, Vblood and Vbrain of calycosin were increased; Cmax blood, AUC0-t blood, AUC0-inf blood, and MRTbrain of 6-gingerol were decreased, CLblood of 6-gingerol was increased. CONCLUSION: This method is simple, rapid, and sensitive. It is suitable for the pharmacokinetic study of Huangqi Guizhi Wuwu Decoction in the blood and brain tissue of rats. Cerebral ischemia-reperfusion injury increased the content of paeoniflorin, calycosin, calycosin-7-o-ß-d-6-glucoside, and 6-gingerol in the blood, affecting the clearance rate of paeoniflorin in the brain, the detention time of calycosin-7-o-ß-d-6-glucoside and the 6-gingerol in the brain. In normal and cerebral ischemia-reperfusion rats, the content of paeoniflorin and 6-gingerol in the blood was higher than that in brain tissue, while the content of calycosin, calycosin-7-o-ß-d-6-glucoside in the brain tissue was higher than that in blood, suggesting that calycosin and calycosin-7-o-ß-d-6-glucoside have brain targeting properties.

Polydatin ameliorates renal injury by attenuating oxidative stress-related inflammatory responses in fructose-induced urate nephropathic mice.

A series of studies have recently demonstrated that the oxidative stress, nuclear factor-kappa B (NF-κB) activation and the subsequent coordinated inflammatory responses played an important role in the pathogenesis of urate nephropathy (UN). Polydatin has been suggested to have the properties of anti-oxidative, anti-inflammatory and nephroprotective effects. However, the possible protective and beneficial effects of polydatin on UN are not fully elucidated. Therefore, we investigated the potential beneficial effects and possible mechanisms of polydatin on UN. In this study, polydatin showed inhibitory activities on xanthine oxidase to repress the level of serum uric acid in vivo and in vitro. Further investigations revealed that polydatin displayed little toxic effects and significantly ameliorated the renal function in fructose-induced UN mice. The nephroprotective activities of polydatin was not only due to the effects on remarkably attenuating the oxidative stress induced by uric acid, but also on markedly suppressing the oxidative stress-related inflammatory cascade, including decreasing the expressions of NF-κB p65, COX-2 and iNOS proteins and inhibiting the productions of TNF-α, PGE(2) and IL-1ß. These findings elucidated that polydatin exhibited prominent nephroprotective activities and low toxic effects.

Chinese medicine formula DSQRL versus glucocorticoids for the treatment of experimental pulmonary fibrosis.

Pulmonary fibrosis (PF) is a devastating interstitial lung disease that may develop idiopathically or as a complication of many other diseases. The outcome of the current main treatment by glucocorticoids is by no means satisfactory. This study has tested a new Chinese medicine Decoction for Strengthening Qi and Replenishing Lung (DSQRL) for the treatment of experimental PF in comparison with prednisolone. Eighty-five rats with PF induced by CCl(4) were randomly divided into 4 groups to undertake treatment either by (a) high dose of prednisolone; (b) Chinese medicine formula DSQRL; (c) combined treatment of the above two; or (d) sham feeding of water in equal volume. At the end of 60 days treatment, the DSQRL treatment achieved a significantly better outcome than prednisolone in terms of general behavior, histological examination, hydroxyproline content of the lung and inflammatory cell counts in bronchoalveolar lavage fluid. Thus, the newly proposed Chinese medicinal formula DSQRL appears to be a better and promising option for PF than glucocorticoids for the treatment of PF.

Polarographic determination of berberine in the presence of H(2)O(2) in medicinal plants.

The mechanism of the parallel catalytic wave of berberine in the presence of H(2)O(2) was studied. The results showed that the reduction process of the C[double bond]N bond of berberine was two successive one-electron transfers, in which an intermediate free radical was involved. When H(2)O(2) was present, it oxidized the free radical to the original C[double bond]N bond, producing the parallel catalytic wave of berberine. In Na(2)B(4)O(7)-Na(2)CO(3) (pH 9.4, 0.08 M)-H(2)O(2)(4 mM) supporting electrolyte, the peak current of the catalytic wave was linear to the berberine concentration in the range of 1.0 x 10(-8)-3.0 x 10(-7) M. The limit of detection was 7.0 x 10(-9) M. The catalytic wave can be applied to direct determination of berberine in medicinal plant Coptis chinensis Franch, after sufficiently diluting the water extraction without preliminary separation.