Showing papers by "Liaoning University of Traditional Chinese Medicine published in 2014"

Potassium Channels: Structures, Diseases, and Modulators

Abstract: Potassium channels participate in many critical biological functions and play important roles in a variety of diseases. In recent years, many significant discoveries have been made which motivate us to review these achievements. The focus of our review is mainly on three aspects. Firstly, we try to summarize the latest developments in structure determinants and regulation mechanism of all types of potassium channels. Secondly, we review some diseases induced by or related to these channels. Thirdly, both qualitative and quantitative approaches are utilized to analyze structural features of modulators of potassium channels. Our analyses further prove that modulators possess some certain natural-product scaffolds. And pharmacokinetic parameters are important properties for organic molecules. Besides, with in silico methods, some features that can be used to differentiate modulators are derived. There is no doubt that all these studies on potassium channels as possible pharmaceutical targets will facilitate future translational research. All the strategies developed in this review could be extended to studies on other ion channels and proteins as well.

Interaction Between Umami Peptide and Taste Receptor T1R1/T1R3

Abstract: The umami taste receptor is a heterodimer composed of two members of the T1R taste receptor family: T1R1 and T1R3. The homology models of the ligand binding domains of the human umami receptor have been constructed based on crystallographic structures of the taste receptor of the central nervous system. Furthermore, the molecular simulations of the ligand binding domain show that the likely conformation was that T1R1 protein exists in the closed conformation, and T1R3 in the open conformation in the heterodimer. The molecular docking study of T1R1 and T1R3 in complex with four peptides, including Lys–Gly–Asp–Glu–Ser–Leu–Leu–Ala, Ser–Glu–Glu, G1u–Ser, and Asp–Glu–Ser, displayed that the amino acid residue of SER146 and Glu277 in T1R3 may play great roles in the synergism of umami taste. This docking result further validated the robustness of the model. In the paper, binding of umami peptide and the T1R1/T1R3 receptor was first described and the interaction is the base of umami activity theory.

Chemical composition and insecticidal activities of the essential oil of Perilla frutescens (L.) Britt. aerial parts against two stored product insects

Abstract: The chemical composition of the essential oil of Perilla frutescens (L.) Britt. aerial parts and its insecticidal activity against Tribolium castaneum and Lasioderma serricorne were investigated. The essential oil of P. frutescens was obtained by hydrodistillation and a total of 34 components in the essential oil were identified with GC–MS. It was found that the main compounds included 2-furyl methyl ketone (71.83 %), decahydro-1-methyl-2-methylene-naphthalene (10.47 %), limonene (5.16 %) and caryophyllene (1.66 %). With a further isolation, the two active constituents were obtained from the essential oil and identified as 2-furyl methyl ketone, limonene. In the progress of assay, it showed that the essential oil and 2-furyl methyl ketone exhibited stronger contact and fumigant activities against the two stored product insects than limonene. Moreover, the essential oil and its constituents exhibited the comparable repellency against the two stored product insects, relative to the positive control, DEET. The results indicate that the essential oil of P. frutescens aerial parts and its isolated compounds have potential for development into natural insecticides or fumigants as well as repellents for control of insects in stored grains.

Bioactivity of Essential Oil of Artemisia argyi Lévl. et Van. and Its Main Compounds Against Lasioderma serricorne

Abstract: Artemisia argyi Levl. et Van., a perennial herb with a strong volatile odor, is widely distrbuted in the world. Essential oil obtained from Artemisia argyi was analyzed by gas chromatography-mass spectrometry (GC-MS). A total of 32 components representing 91.74% of the total oil were identified and the main compounds in the oil were found to be eucalyptol (22.03%), β-pinene (14.53%), β-caryophyllene (9.24%) and (-)-camphor (5.45%). With a further isolation, four active constituents were obtained from the essential oil and identified as eucalyptol, β-pinene, β-caryophyllene and camphor. The essential oil and the four isolated compounds exhibited potential bioactivity against Lasioderma serricorne adults. In the progress of assay, it showed that the essential oil, camphor, eucalyptol, β-caryophyllene and β-pinene exhibited strong contact toxicity against L. serricorne adults with LD50 values of 6.42, 11.30, 15.58, 35.52, and 65.55 μg/adult, respectively. During the fumigant toxicity test, the essential oil, eucalyptol and camphor showed stronger fumigant toxicity against L. serricorne adults than β-pinene (LC50 = 29.03 mg/L air) with LC50 values of 8.04, 5.18 and 2.91 mg/L air. Moreover, the essential oil, eucalyptol, β-pinene and camphor also exhibited the strong repellency against L. serricorne adults, while, β-caryophyllene exhibited attracting activity relative to the positive control, DEET. The study revealed that the bioactivity properties of the essential oil can be attributed to the synergistic effects of its diverse major and minor components. The results indicate that the essential oil of A. argyi and the isolated compounds have potential to be developed into natural insecticides, fumigants or repellents in controlling insects in stored grains and traditional Chinese medicinal materials.

Chondrocyte β-Catenin Signaling Regulates Postnatal Bone Remodeling Through Modulation of Osteoclast Formation in a Murine Model

Abstract: Objective To investigate whether β-catenin signaling in chondrocytes regulates osteoclastogenesis, thereby contributing to postnatal bone growth and bone remodeling.

Selectivity for inhibition of nilotinib on the catalytic activity of human UDP-glucuronosyltransferases

Abstract: 1. Nilotinib, a tyrosine kinase inhibitor, could potently inhibit SN-38 glucuronidation mainly catalyzed by UDP-glucuronosyltransferase (UGT) 1A1. This study was designed to investigate whether nilotinib can be used as a selective inhibitor of UGT1A1 in human liver.2. Assays with recombinant UGTs indicated that nilotinib could strongly inhibit the activity of UGT1A1 and decreased the activity of extra-hepatic UGT1A7 to a much lesser extent. The inhibition on 4-methylumbelliferone (4Mu) glucuronidation by recombinant UGT1A1 obeyed competitive inhibition mechanism, with a kinetic constants (Ki) value of 0.17 μM. Assays with human liver microsomes (HLM) demonstrated that nilotinib could selectively inhibit estradiol-3-O-glucuronidation (E2-3-O-glucuronidation), a probe reaction of UGT1A1. Kinetic studies displayed that the inhibition on E2-3-O-glucuronidation followed non-competitive inhibition model, different from the inhibition on 4Mu glucuronidation. The Ki values were calculated to be 0.14 and 0...

Strategy integrating stepped fragmentation and glycan diagnostic ion-based spectrum refinement for the identification of core fucosylated glycoproteome using mass spectrometry.

Abstract: Core fucosylation (CF) is a special glycosylation pattern of proteins that has a strong relationship with cancer. The Food and Drug Administration (FDA) has approved the core fucosylated α-fetoprotein as a biomarker for the early diagnosis of hepatocellular carcinoma (HCC). The technology for identifying core fucosylated proteins has significant practical value. The major method for core fucosylated glycoprotein/glycopeptide analysis is neutral loss-based MS(3) scanning under collision-induced dissociation (CID) by ion trap mass spectrometry. However, due to the limited speed and low resolution of the MS(3) scan mode, it is difficult to achieve high-throughput, with only dozens of core fucosylated proteins identified in a single run. In this work, we developed a novel strategy for the identification of CF glycopeptides at a large scale, integrating the stepped fragmentation function, one novel feature of quadrupole-orbitrap mass spectrometry, with "glycan diagnostic ion"-based spectrum optimization. By using stepped fragmentation, we were able to obtain both highly accurate glycan and peptide information of a simplified CF glycopeptide in one spectrum. Moreover, the spectrum could be recorded with the same high speed as the conventional MS(2) scan. By using the "glycan diagnostic ion"-based spectrum refinement method, the efficiency of the CF glycopeptide discovery was significantly improved. We demonstrated the feasibility and reproducibility of our method by analyzing CF glycoproteomes of mouse liver tissue and HeLa cell samples spiked with standard CF glycoprotein. In total, 1364 and 856 CF glycopeptides belonging to 702 and 449 CF glycoproteins were identified, respectively, within a 78-min gradient analysis, which was approximately a 7-fold increase in the identification efficiency of CF glycopeptides compared to the currently used method. In this work, we took core fucosylated glycopeptides as a practical example to demonstrate the great potential of our novel method for use in glycoproteome analysis, and we also anticipate using the flexible novel method in other research fields.

Multifunctional targeting daunorubicin plus quinacrine liposomes, modified by wheat germ agglutinin and tamoxifen, for treating brain glioma and glioma stem cells

Abstract: Most anticancer drugs are not able to cross the blood-brain barrier (BBB) effectively while surgery and radiation therapy cannot eradicate brain glioma cells and glioma stem cells (GSCs), hence resulting in poor prognosis with high recurrence rates. In the present study, a kind of multifunctional targeting daunorubicin plus quinacrine liposomes was developed for treating brain glioma and GSCs. Evaluations were performed on in-vitro BBB model, murine glioma cells, GSCs, and GSCs bearing mice. Results showed that the multifunctional targeting daunorubicin plus quinacrine liposomes exhibited evident capabilities in crossing the BBB, in killing glioma cells and GSCs and in diminishing brain glioma in mice. Action mechanism studies indicated that the enhanced efficacy of the multifunctional targeting drugs-loaded liposomes could be due to the following aspects: evading the rapid elimination from blood circulation; crossing the BBB effectively; improving drug uptake by glioma cells and GSCs; down-regulating the overexpressed ABC transporters; inducing apoptosis of GSCs via up-regulating apoptotic receptor/ligand (Fas/Fasl), activating apoptotic enzymes (caspases 8, 9 and 3), activating pro-apoptotic proteins (Bax and Bok), activating tumor suppressor protein (P53) and suppressing anti-apoptotic proteins (Bcl-2 and Mcl-1). In conclusion, the multifunctional targeting daunorubicin plus quinacrine liposomes could be used as a potential therapy for treating brain glioma and GSCs.

Influence of glucocorticoids on the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells

Abstract: Glucocorticoid has been used extensively in clinical applications, because of its several pharmacologic actions, which include immunosuppression, anti-inflammation, anti-shock, and relief of asthma. However, the long-term or high-dose application of glucocorticoid can induce adverse effects such as osteoporosis, which is known in this case as glucocorticoid-induced osteoporosis (GIOP). It is a secondary osteoporosis that results in easy fracturing, and even disability. Therefore it became a thorny issue. The rat model of glucocorticoid-induced osteoporosis (GIOP) was replicated to isolate BMSCs. Rats were assigned into four groups: normal, normal induction, GIOP, and GIOP induction. The growth cycle was monitored by using flow cytometry. Osteogenic differentiation was compared by using alkaline phosphatase (ALP) staining with a modified calcium cobalt method. The quantitative detection of osteoprotegerin and the receptor activator of nuclear factor kappa-B ligand (RANKL) was conducted by using enzyme-linked immunoassay. Finally, renal Klotho mRNA expression was assessed by using RT-PCR. BMSC proliferation was reduced in GIOP rats. The ALP-positive expression of normal BMSCs to the osteogenic induction solution was stronger than that of BMSCs from GIOP rats (P < 0.01). Osteoprotegerin expression was significantly higher in the normal induction group than in the normal, GIOP (P < 0.01), and GIOP induction groups (P < 0.05). RANKL expression was significantly higher in the normal induction group than in the other groups (P < 0.01) and significantly higher in the normal group than in the GIOP and GIOP induction groups (P < 0.01). RT-PCR analysis showed that renal Klotho mRNA expression was significantly reduced in the GIOP group compared with the normal group (P < 0.01). BMSC proliferation, osteogenic differentiation, and reactive activity to an osteogenic inductor were reduced in GIOP rats. Klotho mRNA expression decreased during GIOP induction.

Streptozotocin-induced diabetes increases amyloid plaque deposition in AD transgenic mice through modulating AGEs/RAGE/NF-κB pathway.

Abstract: Background: An increasing number of studies have demonstrated of that diabetes mellitus (DM) is associated with an increased prevalence of Alzheimer disease (AD), the underlying mechanisms are still obscure. Methods: We developed a streptozotocin (STZ)-induced diabetic AD transgenic mouse model and evaluated the effect of hyperglycemia on senile plaque formation. Results: Our data showed that administration of STZ increased the level of blood glucose and increased the advanced glycation end products (AGEs) in brain tissue, and further enhanced the expression levels of the receptor for AGEs (RAGE) and the nuclear factor-kappa B (NF-κB) in the brain, and accelerated the senile plaque formation in the transgenic mice. Our results showed that STZ-induced insulin-deficient hyperglycemia caused the pathophysiology of AD in APP/PS1 transgenic mice by modulating the AGEs/RAGE/NF-κB pathway. Conclusions: Our study suggests that there is a close linkage of DM and cerebral amyloidosis in the pathogenesis of AD.

Multivalent hydrazide-functionalized magnetic nanoparticles for glycopeptide enrichment and identification

Abstract: Among the common approaches for global glycopeptide enrichment, hydrazide chemistry is well recognized. However, conventional hydrazide-functionalized products are composed of a single layer of hydrazide functional groups. Due to the limited specific surface area of such a structure, the loading amount of hydrazide groups immobilized on these materials is restricted. Therefore, these materials can only provide a limited reaction rate with glycopeptides in complex protein samples, which is exacerbated by the microheterogeneities of glycosylation. Here, we introduce a new functionalized magnetic nanoparticle coating with hydrazide-modified non-crosslinked polymer chains. The multivalent hydrazide-functionalized particles were synthesized by the surface-initiated atom transfer radical polymerization (SI-ATRP) technique. The density of the hydrazide groups on the surface of these nanoparticles was three-fold higher than that of conventional single-layered materials. The new particles enabled the highly sensitive and selective enrichment of glycopeptides from a digestion mixture of fetuin, even from a background mixture of non-glycosylated protein that was 100-fold more abundant. The recovery ratio of glycopeptides was determined to be 77.8%, and the glycopeptide binding capacity of the materials was determined to be 25 μg mg−1. Finally, the novel multivalent hydrazide-functionalized particles were applied in the enrichment of N-linked glycopeptides from mouse liver tissues, which resulted in the assignment of 511 unique glycopeptides belonging to 372 different glycoproteins. The results further demonstrated the potential of the multivalent particles for glycopeptide enrichment in complex proteomics samples.

Chemical Constituents and Insecticidal Activities of the Essential Oil of Cinnamomum camphora Leaves against Lasioderma serricorne

Abstract: During our screening program for agrochemicals from Chinese medicinal herbs and wild plants, the essential oil of Cinnamomum camphora leaves was found to possess strong fumigant and contact toxicity against Lasioderma serricorne adults with LC50/LD50 values of 2.5 mg/L air and 21.25 μg/adult, respectively. The essential oil obtained by hydrodistillation was investigated by GC and GC-MS. The main components of the essential oil were identified to be D-camphor (40.54%), linalool (22.92%), cineole (11.26%), and 3,7,11-trimethyl-3-hydroxy-6,10-dodecadien-1-yl acetate (4.50%). Bioactivity-directed chromatographic separation on repeated silica gel columns led to the isolation of D-camphor and linalool. D-camphor and linalool showed strong fumigant toxicity (LC50 = 2.36 and 18.04 mg/L air, resp.) and contact toxicity (LD50 = 13.44 and 12.74 μg/adult, resp.) against L. serricorne. The results indicate that the essential oil of C. camphora and its active compounds had the potential to be developed as natural fumigants and insecticides for control of L. serricorne.

Basic Mechanism Leading to Stimulation of Glycogenolysis by Isoproterenol, EGF, Elevated Extracellular K+ Concentrations, or GABA

Abstract: Glycogenolysis, in brain parenchyma an astrocyte-specific process, has changed from being envisaged as an emergency procedure to playing central roles during brain response to whisker stimulation, memory formation, astrocytic K+ uptake and stimulated release of ATP. It is activated by several transmitters and by even very small increases in extracellular K+ concentration, and to be critically dependent upon an increase in free cytosolic Ca2+ concentration ([Ca2+]i), whereas cAMP plays only a facilitatory role together with increased [Ca2+]i. Detailed knowledge about the signaling pathways eliciting glycogenolysis is therefore of interest and was investigated in the present study in well differentiated cultures of mouse astrocytes. The β-adrenergic agonist isoproterenol stimulated glycogenolysis by a β1-adrenergic effect, which initiated a pathway in which cAMP/protein kinase A activated a Gi/Gs shift, leading to Ca2+-activated glycogenolysis. Inhibition of this pathway downstream of cAMP but upstream of the Gi/Gs shift abolished the glycogenolysis. However, inhibitors operating downstream of the Ca2+-sensitive step, but preventing transactivation-mediated epidermal growth factor (EGF) receptor stimulation, a later step in the activated pathway, also caused inhibition of glycogenolysis. For this reason the effect of EGF was investigated and it was found to be glycogenolytic. Large increases in extracellular K+ activated glycogenolysis by a nifedipine-inhibited L-channel opening allowing influx of Ca2+, known to be glycogenolysis-dependent. Small increases (addition of 5 mM KCl) caused a smaller effect by a similarly glycogenolysis-reliant opening of an IP3 receptor-dependent ouabain signaling pathway. The same pathway could be activated by GABA (also in brain slices) due to its depolarizing effect in astrocytes.

Metabolomics Coupled with Multivariate Data and Pathway Analysis on Potential Biomarkers in Gastric Ulcer and Intervention Effects of Corydalis yanhusuo Alkaloid

Abstract: Metabolomics, the systematic analysis of potential metabolites in a biological specimen, has been increasingly applied to discovering biomarkers, identifying perturbed pathways, measuring therapeutic targets, and discovering new drugs. By analyzing and verifying the significant difference in metabolic profiles and changes of metabolite biomarkers, metabolomics enables us to better understand substance metabolic pathways which can clarify the mechanism of Traditional Chinese Medicines (TCM). Corydalis yanhusuo alkaloid (CA) is a major component of Qizhiweitong (QZWT) prescription which has been used for treating gastric ulcer for centuries and its mechanism remains unclear completely. Metabolite profiling was performed by high-performance liquid chromatography combined with time-of-flight mass spectrometry (HPLC/ESI-TOF-MS) and in conjunction with multivariate data analysis and pathway analysis. The statistic software Mass Profiller Prossional (MPP) and statistic method including ANOVA and principal component analysis (PCA) were used for discovering novel potential biomarkers to clarify mechanism of CA in treating acid injected rats with gastric ulcer. The changes in metabolic profiling were restored to their base-line values after CA treatment according to the PCA score plots. Ten different potential biomarkers and seven key metabolic pathways contributing to the treatment of gastric ulcer were discovered and identified. Among the pathways, sphingophospholipid metabolism and fatty acid metabolism related network were acutely perturbed. Quantitative real time polymerase chain reaction (RT-PCR) analysis were performed to evaluate the expression of genes related to the two pathways for verifying the above results. The results show that changed biomarkers and pathways may provide evidence to insight into drug action mechanisms and enable us to increase research productivity toward metabolomics drug discovery.

Effects of vitexin-2"-O-rhamnoside and vitexin-4"-O-glucoside on growth and oxidative stress-induced cell apoptosis of human adipose-derived stem cells.

Abstract: Objectives Vitexin-2″-O-rhamnoside (VOR) and vitexin-4″-O-glucoside (VOG) are the two main flavonoid glycosides of the leaves of Cratagus pinnatifida Bge var major N E Br that has been widely used for the treatment of cardiovascular system diseases In this study, we simultaneously investigated the influence of VOR and VOG on human adipose-derived stem cells (hADSCs) injury induced by hydrogen peroxide (H2O2) to further characterize their anti-oxidative and anti-apoptotic activity Methods hADSCs were isolated, cultured in vitro and pretreated with 625 μm VOR or 120 μm VOG for 24 h and then exposed to 500 μm H2O2 for an additional 4 h Key findings Pretreatment of hADSCs with VOR and VOG was demonstrated to significantly ameliorate the toxicity and apoptosis effects, such as morphological distortion, nuclear condensation, decreased intracellular caspase-3 activity and percentage of cells in apoptosis/necrosis by using morphological assay, immunocytochemistry and flow cytometric evaluation In addition, VOR and VOG caused no cytotoxic effect on hADSCs at concentrations up to 250 and 480 μm, respectively Conclusions Our results indicated that both VOR and VOG contribute to the protection against H2O2-mediated oxidative stress damage and could be safely used for a wide range of concentrations

Activation of β-catenin signalling leads to temporomandibular joint defects.

Abstract: Despite extensive research in knee and hip osteoarthritis (OA), the underlying mechanism of temporomandibular joint (TMJ) disorder remains largely unknown. The purpose of this study was to determine whether the constitutive activation of β-catenin in the middle and deep layers of the articular cartilage can compromise the homeostasis of this tissue in the TMJ. Col2CreERT2 transgenic mice were bred with RosamT/mG reporter mice to determine Cre recombination efficiency. Col2CreERT2 mice were then crossed with β-cateninflox(ex3)+ mice to generate β-catenin conditional activation mice, β-catenin(ex3)Col2ER. TMJ samples were harvested when the mice were 1-, 3- or 6-month-old and evaluated using histology, histomorphometry and immunohistochemistry. β-catenin(ex3)Col2ER mice were further crossed with Mmp13flox/flox and Adamts5-/- mice to generate (β-catenin(ex3)/Mmp13)Col2ER and β-catenin(ex3)Col2ER)/Adamts5-/- double mutant mice to investigate the role of Mmp13 and Adamts5 in the development of TMJ disorder. High levels of Cre-recombination were seen in Col2CreERT2;RosamT/mGmice. Progressive TMJ defects developed in 1-, 3- and 6-month-old β-catenin(ex3)Col2ER mice, as revealed by histology and histomorphometry. Results further demonstrated that the defects observed in β-catenin(ex3)Col2ER mice were significantly decelerated after deletion of the Mmp13 or Adamts5 gene in (β-catenin(ex3)/Mmp13)Col2ER or β-catenin(ex3)Col2ER/Adamts5-/- double mutant mice. In summary, we found that β-catenin is a critical gene in the induction of TMJ cartilage degeneration, and over-expressing β-catenin in TMJ cartilage leads to defects assembling an OA-like phenotype. Deletion of Mmp13 and Adamts5 in β-catenin(ex3)Col2ER mice ameliorates the development of TMJ defects. This study suggests that Mmp13 and Adamts5 could be potential therapeutic targets for the treatment of TMJ disorders.

The impact of CYP2D6 polymorphisms on the pharmacokinetics of codeine and its metabolites in Mongolian Chinese subjects

Abstract: Codeine is an analgesic drug acting on μ-opioid receptors predominantly via its metabolite morphine formed almost exclusively by CYP2D6. Genetic polymorphisms in CYP2D6 are associated with diminished pain relief and/or severe opioid side effects. In Chinese individuals, CYP2D6*10 is the most common allele with reduced enzyme activity. In this study, we investigated the effect of this allele on the pharmacokinetics of codeine and its metabolites. A blood sample was collected from healthy Mongolian volunteers for CYP2D6 genotyping using a PCR-RFLP assay. A pharmacokinetic study was then carried out in three groups with CYP2D6*1/*1 (n = 10), CYP2D6*1/*10 (n = 10) and CYP2D6*10/*10 (n = 9) genotypes by collecting serial blood samples for determination of plasma levels of codeine and its metabolites, morphine, morphine 3-glucuronide (M3G) and morphine 6-glucuronide (M6G) before and after a single 30-mg oral dose of codeine phosphate. Codeine and its metabolites were measured by LC-MS/MS. No significant differences were observed in the pharmacokinetic parameters of codeine in the three genotype groups. However, the C max and AUC0-∞ of morphine, M3G and M6G were significantly different between the study groups (P < 0.05). Compared with the *1/*1 group, the AUC0-∞ for morphine in the *1/*10 and *10/*10 groups decreased by ratios (95 % CI) of 0.93 (0.26–1.59) and 0.494 (0.135–0.853) respectively. Corresponding ratios for M3G were 0.791 (0.294–1.288) and 0.615 (0.412–0.818) and for M6G were 0.643 (0.39–0.957) and 0.423 (0.267–0.579). This study demonstrates that the CYP2D6*10 allele plays an important role in the pharmacokinetics of the O-demethylated metabolites of codeine after oral administration.

Anti-inflammatory and anti-nociceptive activities of methanol extract from aerial part of Phlomis younghusbandii Mukerjee.

Abstract: This study was designed to investigate the anti-inflammatory and anti-nociceptive activity of the methanol extract from the aerial part of Phlomis younghusbandii (MEAP) and to explore the possible related mechanisms. Anti-inflammatory effects of MEAP were evaluated by using the ear edema test induced by dimethylbenzene and vascular permeability test induced by acetic acid. Anti-nociceptive activities of MEAP were evaluated by the chemical nociception in models of acetic acid-induced writhing and formalin-induced hind paw licking, and by the thermal nociception in hot plate tests. Mechanisms of MEAP activities also were explored by evaluating expression levels of TNF-α, IL-6 and iNOS induced by LPS using real-time fluorogenic PCR and expression of COX-2 using Western blotting and an open-field test. The results indicated that the MEAP administered orally could significantly decrease ear edema induced by dimethylbenzene and increase vascular permeability induced by acetic acid. Additionally, the nociceptions induced by acetic acid and formalin were significantly inhibited. The anti-nociceptive effect could not be decreased by naloxone in the formalin test, and MEAP did not affect the normal autonomic activities of mice. Expression levels of pro-inflammatory cytokines (TNF-α, IL-6, iNOS) induced by LPS were decreased obviously by treatment with MEAP. Furthermore, COX-2 expression in the spinal dorsal horns of the pain model mice induced by formalin was significantly down-regulated by MEAP. In conclusion, MEAP has significant anti-inflammatory and antinociceptive activities, and the mechanisms may be related to the down-regulated expression of TNF-α, IL-6, iNOS and COX-2.

Hepatic, gastric, and intestinal first-pass effects of vitexin in rats.

Abstract: Context: Recent research has demonstrated that vitexin exhibits a prominent first-pass effect In this light, it is necessary to investigate the causes of this distinct first-pass effectObjective: The aim of this study was to evaluate hepatic, gastric, and intestinal first-pass effects of vitexin in rats and, furthermore, to investigate the role of P-glycoprotein (P-gp) and cytochrome P450 3A (CYP3A) in the absorption and secretion of vitexin in the duodenumMaterials and methods: Vitexin was infused into rats intravenously, intraportally, intraduodenally, and intragastrically (30 mg/kg) In addition, verapamil (50 mg/kg), a common substrate/inhibitor of P-gp and CYP3A, was also instilled with vitexin into the duodenum to investigate the regulatory action of P-gp and CYP3A The plasma concentrations of vitexin were measured by the HPLC method using hesperidin as an internal standardResults: The hepatic, gastric, and intestinal first-pass effects of vitexin in rats were 52%, 313%, and 941%, re

Role of moxibustion in inflammatory responses during treatment of rat ulcerative colitis

Abstract: AIM: To investigate the efficacy of moxibustion in ulcerative colitis (UC) rats from morphological, immunological and molecular biological perspectives. METHODS: Thirty-two Sprague-Dawley rats were randomly assigned to a blank control group (normal rats, n = 6) and a model replication (MR) group (UC rats, n = 26). A UC model was established by 2,4,6-trinitrobenzenesulfonic acid/dextran sulfate sodium enema. Rats in the MR group were further randomly assigned to a 9-min moxibustion (9M) group (9 moxa-cone, n = 6), 6-min moxibustion (6M) group (6 moxa-cone, n = 6), 3-min moxibustion (3M) group (3 moxa-cone, n = 6), and a waiting list control (WLC) group (no moxibustion treatment, n = 6). Rats in the moxibustion treatment group were treated in 14 sessions over 28 d. Disease activity, local tissue morphology, serum level of interleukin (IL)-8 and IL-10, and expression of Toll-like receptor (TLR)9 as well as nuclear factor (NF)-κB p65 in colonic tissue were determined by disease activity index (DAI), hematoxylin and eosin staining, electron microscopy, enzyme-linked immunosorbent assay and Western blotting, respectively. RESULTS: DAI was lowest in the 9M group and highest in the WLC group. The differences in DAI between the moxibustion treatment (3M, 6M, 9M) and no treatment groups were significant for all one-to-one comparisons (0.60 ± 0.54 vs 1.20 ± 0.44, 0.60 ± 0.54 vs 1.80 ± 0.45, 0.60 ± 0.54 vs 3.0 ± 0.45, respectively, P < 0.05). Light and electron microscopy showed that the neatness of the glandular arrangement in colonic mucosal epithelia gradually increased in the WLC, 3M, 6M to 9M groups. IL-8 level successively decreased while IL-10 level increased from the WLC to 3M, 6M and 9M groups. The differences among these groups were significant for all comparisons (105.46 ± 8.75 vs 76.61 ± 3.58, 105.46 ± 8.75 vs 69.78 ± 1.87, 105.46 ± 8.75 vs 67.41 ± 1.84, respectively, P < 0.01 for IL-8; and 30.83 ± 1.29 vs 75.64 ± 1.90, 30.83 ± 1.29 vs 80.90 ± 3.16, 30.83 ± 1.29 vs 83.46 ± 2.37, respectively, P < 0.01 for IL-10), except comparison of 6M vs 9M. Expression of TLR9 and NF-κB p65 decreased in order: highest in the WLC group and lowest in the 9M group. In addition, the differences among the WLC, 3M, 6M and 9M groups were significant for all comparisons (0.492 ± 0.026 vs 0.380 ± 0.022, 0.492 ± 0.026 vs 0.355 ± 0.005, 0.492 ± 0.026 vs 0.327 ± 0.015, respectively, P < 0.05 for TLR9; and 0.436 ± 0.041 vs 0.326 ± 0.022, 0.436 ± 0.041 vs 0.293 ± 0.006, 0.436 ± 0.041 vs 0.265 ± 0.017, respectively, P < 0.05 for NF-κB p65). CONCLUSION: Moxibustion repairs damaged colonic mucosa, suppresses serum IL-8, activates serum IL-10 level, and decreases expression of TLR-9 and NF-κB p65 in UC rats.

Evidence for the involvement of JAK/STAT/SOCS pathway in the mechanism of Tangshen formula-treated diabetic nephropathy.

Abstract: Diabetic nephropathy is one of the most significant microvascular complications associated with diabetes. Until now, there is no effective treatment and the gene mechanism of diabetic nephropathy is still unclear. Tangshen formula is a traditional Chinese medicine, and has been shown to have good clinical efficacy in diabetic nephropathy treatment. The objective of this study was to investigate the changes of gene expression profiling and explore the molecular mechanism using a db/db mice model treated by Tangshen formula. After administration for 12 weeks, a microarray was applied to detect the gene expression of db/db mice kidney tissues. Quantitative real-time PCR was used to confirm the differential gene expression and carry out a JAK/STAT/SOCS signaling pathway study. Treatment with Tangshen formula reduced the levels of serum glucose and urinary albumin in db/db mice, and the effects of Tangshen formula on db/db mice were significantly different from the positive control (Losartan potassium tablets) on microarray data. It also showed that the JAK/STAT/SOCS signaling pathway played an important role in the treatment process. The expressions of JAK1, JAK2, and STAT3 were upregulated, and STAT4 was downregulated in Tangshen formula-treated db/db mice. SOCS1, 3, and 7 were all activated, while negative feedback regulated other related genes in the JAK/STAT/SOCS pathway. Our study suggested that Tangshen formula has beneficial effects on diabetic nephropathy treatment via regulating the JAK/STAT/SOCS signaling pathway. This study will help to provide evidence-based recommendations for Tangshen formula clinical treatment.

Chemical constituents and biological activities against Tribolium castaneum (Herbst) of the essential oil from Citrus wilsonii leaves

Abstract: The essential oil obtained from Citrus wilsonii Tanaka leaves by hydrodistillation was investigated by GC and GC-MS. The main components of the essential oil were identified to be citronellol (16.94 %), neryl acetate (10.35 %), γ-terpinene (9.85 %), citronellal (9.36 %) and β-pinene (6.72 %). These four compounds, predicted with a bioactivity-test to be the active constituents, were isolated and identified. It was found that the essential oil of C. wilsonii leaves and the isolated compounds possessed fumigant and contact toxicity against Tribolium castaneum adults. The essential oil and γ-terpinene showed strong fumigant toxicity against T. castaneum (LC50 = 8.18 and 4.09 mg L -1 , respectively). The repellency of the crude oil and the active compounds was also determined. Citronellol, neryl acetate and β-pinene were strongly repellent (100, 86 and 92 %, respectively, at 78.63 nL cm -2 , after 2 h treatment) against T. castaneum. The essential oil and citronellol exhibited the same level of repellency compared with the positive control, N,N-diethyl-meta-toluamide (DEET, N,N-diethyl-3-methylbenzamide). The results indicate that the essen- tial oil of C. wilsonii leaves and its active compounds had the potential to be developed as natural fumigants, insecticides and repellents for the control of T. castaneum.