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1.
Zhongguo Zhong Yao Za Zhi ; 49(1): 62-69, 2024 Jan.
Article in Chinese | MEDLINE | ID: mdl-38403339

ABSTRACT

The volatile oils are the effective components of Agastache rugosa, which are stored in the glandular scale. The leaves of pulegone-type A. rugosa were used as materials to observe the leaf morphology of A. rugosa at different growth stages, and the components of volatile oils in gland scales were detected by GC-MS. At the same time, qRT-PCR was used to determine the relative expression of key enzyme genes in the biosynthesis pathway of monoterpenes in volatile oils. The results showed that the density of A. rugosa glandular scale decreased first and then tended to be stable. With the growth of leaves, the relative content of pulegone decreased from 79.26% to 3.94%(89.97%-41.44%), while that of isomenthone increased from 2.43% to 77.87%(0.74%-51.01%), and the changes of other components were relatively insignificant. The correlation analysis between the relative content of monoterpenes and the relative expression levels of their key enzyme genes showed that there was a significant correlation between the relative content of menthone and isomenthone and the relative expression levels of pulegone reductase(PR)(r>0.6, P<0.01). To sum up, this study revealed the accumulation rules of the main components of the contents of the glandular scale of A. rugosa and the expression rules of the key enzyme genes for biosynthesis, which provided a scientific basis and data support for determining the appropriate harvesting period and quality control of the medicinal herbs. This study also initially revealed the biosynthesis mechanism of the monoterpenes mainly composed of pulegone and isomenthone in A. rugosa, laying a foundation for further research on the molecular mechanism of synthesis and accumulation of monoterpenes in A. rugosa.


Subject(s)
Agastache , Cyclohexane Monoterpenes , Oils, Volatile , Oils, Volatile/analysis , Agastache/metabolism , Monoterpenes/metabolism
2.
Zhongguo Zhong Yao Za Zhi ; 48(22): 6039-6050, 2023 Nov.
Article in Chinese | MEDLINE | ID: mdl-38114210

ABSTRACT

Terpenoids are important secondary metabolites of plants that possess both pharmacological activity and economic value. Terpene synthases(TPSs) are key enzymes in the synthesis process of terpenoids. In order to investigate the TPS gene family members and their potential functions in Schizonepeta tenuifolia, this study conducted a systematic analysis of the TPS gene family of S. tenuifolia based on the whole genome data of S. tenuifolia using bioinformatics methods. The results revealed 57 StTPS members identified from the genome database of S. tenuifolia. The StTPS family members encoded 285-819 amino acids, with protein molecular weights ranging from 32.75 to 94.11 kDa, all of which were hydrophilic proteins. The StTPS family members were mainly distributed in the cytoplasm and chloroplasts, exhibiting a random and uneven physical localization pattern. Phylogenetic analysis showed that the StTPS genes family were divided into six subgroups, mainly belonging to the TPS-a and TPS-b subfamilies. Promoter analysis predicted that the TPS gene family members could respond to various stressors such as light, abscisic acid, and methyl jasmonate(MeJA). Transcriptome data analysis revealed that most of the TPS genes were expressed in the roots of S. tenuifolia, and qRT-PCR analysis was conducted on genes with high expression in leaves and low expression in roots. Through the analysis of the TPS gene family of S. tenuifolia, this study identified StTPS5, StTPS18, StTPS32, and StTPS45 as potential genes involved in sesquiterpene synthesis of S. tenuifolia. StTPS45 was cloned for the construction of an prokaryotic expression vector, providing a reference for further investigation of the function and role of the TPS gene family in sesquiterpene synthesis.


Subject(s)
Lamiaceae , Sesquiterpenes , Phylogeny , Terpenes/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Lamiaceae/genetics
3.
Chemosphere ; 339: 139694, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37536538

ABSTRACT

In order to remove high concentrations of ammonia nitrogen (NH4+-N) and refractory sulfamethazine (SM2) from swine digestion effluent, different carbon/nitrogen (C/N) ratios and salinity were used to determine the effects of pollutants removal in the microalgae biofilm system. Microalgae biofilm treatment under optimal environmental conditions in synthetic swine digestion effluent were C/N ratio of 20 and salinity of 140 mM. In order to make the actual swine digestion effluent discharge up to the standard, three different two-cycle treatments (suspended microalgae, microalgae biofilm, microalgae biofilm under the optimal conditions) were studied. The results showed that after two-cycle treatment with microalgae biofilm under the optimal conditions, the actual swine digestion effluent levels of total nitrogen (TN), NH4+-N, total phosphorus (TP), chemical oxygen demand (COD), SM2 were 22.65, 9.32, 4.11, 367.28, and 0.99 mg L-1, respectively, which could satisfy the discharge standards for livestock and poultry wastewater in China. At the same time, first-order kinetic simulation equations suggested a degradation half-life of 4.85 d for SM2 under optimal conditions in microalgae biofilm, and microbial community analysis indicated that the dominant genus was Halomonas. Furthermore, 35.66% of lipid, 32.56% of protein and 18.44% of polysaccharides were harvested after two-cycle in microalgae biofilm treatment under optimal environmental conditions. These results indicated that the regulation of C/N and salinity in microalgae biofilm for the treatment of swine digestion effluent was a high-efficiency strategy to simultaneously achieve wastewater treatment and bioenergy production.


Subject(s)
Microalgae , Nitrogen , Animals , Swine , Nitrogen/metabolism , Microalgae/metabolism , Carbon/metabolism , Salinity , Biofilms , Phosphorus/metabolism , Digestion , Biomass
4.
Ying Yong Sheng Tai Xue Bao ; 33(10): 2611-2618, 2022 Oct.
Article in Chinese | MEDLINE | ID: mdl-36384594

ABSTRACT

As an important parameter regulating soil carbon mineralization, microbial carbon use efficiency (CUE) is essential for the understanding of carbon (C) cycle in terrestrial ecosystems. Three nitrogen supplemental levels, including control (0 kg N·hm-2·a-1), low nitrogen (40 kg N·hm-2·a-1), and high nitrogen (80 kg N·hm-2·a-1), were set up in a Castanopsis fabri forest in the Daiyun Mountain. The basic physical and chemical properties, organic carbon fractions, microbial biomass, and enzyme activities of the soil surface layer (0-10 cm) were measured. To examine the effects of increasing N deposition on microbial CUE and its influencing factors, soil microbial CUE was measured by the 18O-labelled-water approach. The results showed that short-term N addition significantly reduced microbial respiration rate and the activities of C and N acquisition enzymes, but significantly increased soil microbial CUE. ß-N-acetyl amino acid glucosidase (NAG)/microbial biomass carbon (MBC), microbial respiration rate, ß-glucosidase (BG)/MBC, cellulose hydrolase (CBH)/MBC, and soil organic carbon content were the main factors affecting CUE. Moreover, CUE significantly and negatively correlated with NAG/MBC, microbial respiration rate, BG/MBC, and CBH/MBC, but significantly and positively correlated with soil organic carbon. In summary, short-term N addition reduced the cost of soil microbial acquisition of C and N and microbial respiration, and thus increased soil microbial CUE, which would increase soil carbon sequestration potential of the C. fabri forest.


Subject(s)
Carbon , Soil , Soil/chemistry , Nitrogen/analysis , Soil Microbiology , Ecosystem , Forests
5.
Ying Yong Sheng Tai Xue Bao ; 33(8): 2178-2186, 2022 Aug.
Article in Chinese | MEDLINE | ID: mdl-36043825

ABSTRACT

Soil phosphatases are important in the mineralization of organophosphates and in the phosphorus (P) cycle. The kinetic mechanisms of phosphatases in response to nitrogen (N) deposition remain unclear. We carried out a field experiment with four different concentrations of N: 0 g N·hm-2·a-1(control), 20 g N·hm-2·a-1(low N), 40 g N·hm-2·a-1(medium N), and 80 g N·hm-2·a-1(high N) in a subtropical Moso bamboo forest. Soil samples were then collected from 0 to 15 cm depth, after 3, 5 and 7 years of N addition. We analyzed soil chemical properties and microbial biomass. Acid phosphatase (ACP) was investigated on the basis of maximum reaction velocity (Vm), Michaelis constant (Km), and catalytic efficiency (Ka). Results showed that N addition significantly decreased soil dissolved organic carbon (DOC), available phosphorus, and organophosphate content, but significantly increased soil ammonium, nitrate-N content, and Vm. There was a significant relationship between Vm and the concentrations of available phosphorus, organophosphate, and soil DOC. In general, N addition substantially increased Ka, but did not affect Km. The Km value in the high N treatment group was higher than that in the control group after five years of N addition. Km was significantly negatively associated with both available phosphorus and organophosphate. Medium and high N treatments had stronger effects on the kinetic parameters of ACP than low N treatment. Results of variation partition analysis showed that changes in soil chemical properties, rather than microbial biomass, dominated changes in Vm(47%) and Km(33%). In summary, N addition significantly affected substrate availability in Moso bamboo forest soil and modulated soil P cycle by regulating ACP kinetic parameters (especially Vm). The study would improve the understanding of the mechanisms underlying soil microorganisms-regulated soil P cycle under N enrichment. These mechanisms would identify the important parameters for improving soil P cycling models under global change scenarios.


Subject(s)
Nitrogen , Soil , Acid Phosphatase , Carbon/analysis , China , Forests , Nitrogen/analysis , Organophosphates , Phosphoric Monoester Hydrolases , Phosphorus/analysis , Poaceae , Soil/chemistry , Soil Microbiology
6.
Ying Yong Sheng Tai Xue Bao ; 33(1): 33-41, 2022 Jan.
Article in Chinese | MEDLINE | ID: mdl-35224923

ABSTRACT

Understanding changes in soil enzyme activities and ecoenzymatic stoichiometry is important for assessing soil nutrient availability and microbial nutrient limitation in mountain ecosystems. However, the variations of soil microbial nutrient limitation across elevational gradients and its driving factors in subtropical mountain forests are still unclear. In this study, we measured soil properties, microbial biomass, and enzyme activities related to carbon (C), nitrogen (N), and phosphorus (P) cycling in Pinus taiwanensis forests at different altitudes of Wuyi Mountains. By analyzing the enzyme stoichiometric ratio, vector length (VL), and vector angle (VA), the relative energy and nutrient limitation of soil microorganisms and its key regulatory factors were explored. The results showed that ß-glucosaminidase (BG) activities increased along the elevational gradient, while the activities of ß-N-acetyl glucosaminidase (NAG), leucine aminopeptidase (LAP), acid phosphatase (AcP) and (NAG+LAP)/microbial biomass carbon (MBC) and AcP/MBC showed the opposite trend. Enzyme C/N, enzyme C/P, enzyme N/P, and VL were enhanced with increasing elevation, while VA decreased, indicating a higher degree of microbial P limitation at low elevation and higher C limitation at high elevation. In addition, our results suggested that dissolved organic carbon and microbial biomass phosphorus are critical factors affecting the relative energy and nutrient limitation of soil microorganisms at different elevations. The results would provide a theoretical basis for the responses of soil carbon, nitrogen, and phosphorus availability as well as the relative limitation of microbial energy and nutrition to elevational gradients, and improve our understanding of soil biogeochemical cycle process in subtropical montane forest ecosystems.


Subject(s)
Pinus , Soil , Carbon/analysis , China , Ecosystem , Forests , Nitrogen/analysis , Phosphorus/analysis , Soil Microbiology
7.
Article in Chinese | WPRIM | ID: wpr-940711

ABSTRACT

ObjectiveTo explore the potential mechanism of Polygonati Rhizoma on the treatment of osteoporosis (OP) based on network pharmacology and molecular docking method and to verify the mechanism by experiments. MethodThe main active ingredients and corresponding targets of Polygonati Rhizoma were screened out from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) 2.3 by conditional searching. The treatment targets were obtained from the genes related to OP and DisGeNET 7.0. The potential target genes of Polygonati Rhizoma for treating OP were obtained by the crossing of the corresponding targets and the treatment targets. Cytoscape 3.7.1 was used to construct the “Polygonati Rhizoma-active ingredient-potential target” network. The protein-protein interaction (PPI) analysis was carried out by STRING 11.0, and the PPI network was constructed. Metascape 3.5 was used to conduct Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of the key targets. The core ingredients and key targets of Polygonati Rhizoma were selected for molecular docking by AutoDock Vina 1.1.2. Finally, the effect of β-sitosterol on osteogenic differentiation of MC3T3-E1 cells in rats was observed. ResultTwelve active ingredients and 32 potential targets of Polygonati Rhizoma for OP treatment were screened out. Six active ingredients including baicalein and β-sitosterol and key targets including protein kinase 1 (Akt1), tumor suppressor p53 (TP53), vascular endothelial growth factorA (VEGFA), proto-oncogene Jun (JUN), matrix metalloproteinase-9 (MMP-9), and proto-oncogene c-Fos (FOS) were obtained by Cytoscape 3.7.1 topological analysis. A total of 995 GO entries and 181 signaling pathways involving the response to reactive oxygen species and regulations of growth were obtained from GO and KEGG enrichment analyses. The results of molecular docking showed that the core active ingredients possessed good binding activities with the respective key targets. The results of cell experiments showed that β-sitosterol promoted the osteogenic differentiation at the concentration of 2.5 μmol·L-1 and 5 μmol·L-1. ConclusionPolygonati Rhizoma had the therapeutic effect on treating OP by regulating inflammation, oxidative stress, apoptosis, and metabolism. The β-sitosterol significantly promoted the osteogenic differentiation of MC3T3-E1 cells.

8.
Chinese Journal of Hematology ; (12): 209-214, 2022.
Article in Chinese | WPRIM | ID: wpr-929559

ABSTRACT

Objective: This study aimed to see how different initial treatment regimens affected the long-term prognosis of patients with extranodal marginal zone mucosa-associated lymphoid tissue lymphoma confining to the ocular adnexal (OAML) . Methods: Between April 2008 and April 2019, 109 patients with initial mucosa-associated lymphoid tissue confining to ocular adnexal were evaluated and followed-up, and the prognosis of various initial treatment regimens were examined. Results: A total of 36 patients underwent complete surgical resection of the lesions, and 73 patients had residual lesions after surgery, of which 37 patients chose watchful waiting, and 36 patients chose treatment. The treatment regimen included local radiotherapy and systemic treatment (chemotherapy, immunochemotherapy, the combination of radiotherapy and chemotherapy, etc.) , and no serious toxic and side effects were observed in patients receiving systemic treatment. The median follow-up time was 61 (10-142) months. The 5-year and 10-year progression-free survival (PFS) of monocular involvement patients were 78.2% and 76.0% . The 5-year and 10-year PFS rates of patients with binocular involvement were 64.4% and 23.5%. There was significant diference in PFS between patients with monocular and binocular involvement (P=0.010) . Patients who received additional treatment had higher PFS than those patients in the watchful waiting group (P=0.046) . The 5-year PFS was 71.4% and 90.1% among patients in the watchful waiting group and those who received additional treatment, whereas the 10-year PFS was 63.5% and 75.1% , respectively. Patients with OAML were still a risk of disease progression after 5 years. Conclusions: Patients with binocular involvement OAML at the start of the disease had a poor prognosis, but treatment could reduce the risk of recurrence/progression. Systemic therapy is one of the first-line treatment options for patients with OAML, who require long-term monitoring.


Subject(s)
Humans , Eye Neoplasms/radiotherapy , Lymphoid Tissue/pathology , Lymphoma, B-Cell, Marginal Zone/therapy , Prognosis , Retrospective Studies , Treatment Outcome
9.
J Sci Food Agric ; 101(15): 6424-6433, 2021 Dec.
Article in English | MEDLINE | ID: mdl-33987828

ABSTRACT

BACKGROUND: Peanut milk benefits human health mainly due to its high protein content and suitable amino acid composition. To reveal the molecular mechanism affecting the quality of peanut milk, tandem mass tag (TMT)-labeled proteomic analysis was applied to identify the proteome variation between two peanut cultivars that produced peanut milk with the best and worst stability. RESULTS: A total of 478 differentially abundant proteins (fold change >1.2 or <0.83, P < 0.05) were identified. Most of these proteins were located in the cytoplasm and chloroplasts. Correlation analysis showed that RNA recognition motif (RRM) domain-containing protein (17.1 kDa) had a negative relationship with the sedimentation rate of peanut milk and that 22.0 kDa class IV heat shock protein was negatively correlated with the creaming index (P < 0.05). Bioinformatic analysis showed that the molecular function of RRM domain-containing protein (17.1 kDa) was associated with RNA binding and nucleotide binding, and 22.0 kDa class IV heat shock protein was involved in the pathway of protein processing in the endoplasmic reticulum. CONCLUSION: Overall, the differentially abundant proteins in the biological metabolic pathway might offer some potential markers to guide future peanut breeding, especially for the production of peanut milk. © 2021 Society of Chemical Industry.


Subject(s)
Arachis/chemistry , Plant Preparations/chemistry , Plant Proteins/chemistry , Arachis/classification , Arachis/genetics , Arachis/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Protein Binding , Protein Domains , Protein Stability , Proteomics
10.
Ying Yong Sheng Tai Xue Bao ; 32(2): 521-528, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33650361

ABSTRACT

The activity and stoichiometry of soil extracellular enzyme can provide a good indication for changes in soil nutrient availability and microbial demands for nutrients. However, it remains unclear how would nitrogen (N) deposition affect nutrient limitation of microbes in subtropical forest soils. We conducted a 5 years N addition experiment in a subtropical Phyllostachys pubescens forest. The soil nutrients and enzyme activities associated with carbon (C), N, and phosphorus (P) cycles were measured. We also examined the nutrient distribution of microorganisms using enzyme stoichiometry and vector analysis. The results showed that N addition significantly decreased the contents of soil soluble organic C and available P and increased that of available N. Furthermore, N addition significantly decreased ß-N-acetyl-glucosaminidase (NAG) activity and NAG/ microbial biomass carbon (MBC), and increased acid phosphatase (ACP) and ACP/MBC. The low and moderate N addition levels significantly increased enzyme C/P, vector length, and vector angle, but significantly decreased enzyme N/P. Results of redundancy analysis showed that the change in soil enzyme activity and enzymatic stoichiometry were mainly driven by soil available P content under N addition. In summary, N addition altered the microbial nutrient acquisition strategy, which increased nutrient allocation to P-acquiring enzyme production but reduced that to N-acquiring enzyme production. Moreover, N addition exacerbated the C and P limitation of soil microorganisms. Appropriate amount of P fertilizer could be applied to improve soil fertility of subtropical P. pubescens forest in the future.


Subject(s)
Nitrogen , Phosphorus , Carbon/analysis , China , Forests , Nitrogen/analysis , Phosphorus/analysis , Soil , Soil Microbiology
11.
Carbohydr Polym ; 251: 117060, 2021 Jan 01.
Article in English | MEDLINE | ID: mdl-33142612

ABSTRACT

Due to the poor controlled release ability, bio-based materials are difficult for large scale application on controlled release fertilizers (CRFs). Starch-based polyol (SP) and castor oil (CO) were mutually modified, and a cross-linked polymer film was formed on the surface of urea by in-situ reaction, which improved the slow release ability of the bio-based material. The results showed that increasing the CO ratio reduced porosity of coating and prolonged the nitrogen (N) release period, while the SP changed the high-temperature wrinkle characteristics and regulated the early N release rate. The mutual modification achieved an ultra-long release period of bio-based CRUs for 7 months. The degradation rate during nine months of bio-based coatings (5.05 %) was significantly higher than that of petroleum-based (3.74 %), and the coating was non-toxic to rice seeds. Mutual modification provided a safe and effective solution for the preparation of bio-based CRFs with long-term controlled release capability.


Subject(s)
Castor Oil/chemistry , Cross-Linking Reagents/chemistry , Delayed-Action Preparations/chemistry , Fertilizers/analysis , Polyurethanes/chemistry , Starch/chemistry , Urea/metabolism , Delayed-Action Preparations/pharmacology , Oryza/chemistry , Polymers/chemistry , Urea/chemistry
12.
Ying Yong Sheng Tai Xue Bao ; 31(3): 753-760, 2020 Mar.
Article in Chinese | MEDLINE | ID: mdl-32537969

ABSTRACT

Phosphorus (P) is an important nutrient for plant and microbial growth. Soil P availabi-lity is poor in subtropical areas. Long-term heavy nitrogen (N) deposition might further reduce P availability. The experiment was performed in a Phyllostachys pubescens forest in Daiyun Mountain. The effects of N application on soil basic physical and chemical properties, soil P fractions, microbial biomass, and acid phosphomonoesterase activity were analyzed after three years of N application. The results showed that N application significantly increased NO3--N content and thus soil N availability, while it significantly reduced the percentage of decomposable organic P to total P, with the ratio of carbon (C) to organic P being over 200. The soil microbial biomass C, microbial biomass P, acid phosphomonoesterase, and the ratio of microbial biomass N to microbial biomass P and microbial biomass C to microbial biomass P were increased as the N application rate increased. There was a significant negative correlation between the percentage of decomposable organic P to total P and microbial biomass P. Consequently, N application enhanced soil P limitation and increased microbial P demand.


Subject(s)
Nitrogen , Phosphorus , Biomass , Carbon , China , Forests , Soil , Soil Microbiology
13.
J Agric Food Chem ; 68(24): 6530-6543, 2020 Jun 17.
Article in English | MEDLINE | ID: mdl-32383865

ABSTRACT

This study aimed to investigate the protective mechanism of common buckwheat (Fagopyrum esculentum Moench.) against nonalcoholic fatty liver disease (NAFLD) associated with dyslipidemia in mice that were fed a high-fat and high-cholesterol diet (HFD). Results showed that oral supplementation of common buckwheat significantly improved physiological indexes and biochemical parameters related to dyslipidemia and NAFLD in mice fed with HFD. Furthermore, the HFD-induced reductions in fecal short-chain fatty acids were reversed by common buckwheat intervention, which also increased the fecal bile acid (BA) abundance compared with HFD-induced hyperlipidemic mice. Liver metabolomics based on ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry demonstrated that common buckwheat supplementation made significant regulatory effects on the pentose phosphate pathway, starch and sucrose metabolism, primary BA biosynthesis, and so forth. The results of high-throughput sequencing revealed that common buckwheat supplementation significantly altered the structure of the intestinal microbiota in mice fed with HFD. The correlations between lipid metabolic parameters and intestinal microbial phylotypes were also revealed by the heatmap and network. Additionally, common buckwheat intervention regulated the mRNA expressions of genes responsible for liver lipid metabolism and BA homeostasis, thus promoting BA synthesis and excretion. These findings confirmed that common buckwheat has the outstanding ability of improving lipid metabolism and could be used as a potential functional food for the prevention of NAFLD and hyperlipidemia.


Subject(s)
Cholesterol/metabolism , Fagopyrum/metabolism , Hyperlipidemias/diet therapy , Non-alcoholic Fatty Liver Disease/diet therapy , Animals , Bile Acids and Salts/metabolism , Cholesterol/adverse effects , Diet, High-Fat/adverse effects , Gastrointestinal Microbiome , Humans , Hyperlipidemias/etiology , Hyperlipidemias/metabolism , Hyperlipidemias/microbiology , Liver/metabolism , Male , Mice , Non-alcoholic Fatty Liver Disease/etiology , Non-alcoholic Fatty Liver Disease/metabolism , Non-alcoholic Fatty Liver Disease/microbiology
14.
Front Pharmacol ; 11: 610772, 2020.
Article in English | MEDLINE | ID: mdl-33569004

ABSTRACT

Isorhamnetin (ISO), a naturally occurring plant flavonoid, is widely used as a phytomedicine. The major treatment modality for non-small-cell lung carcinoma (NSCLC) is radiotherapy. However, radiotherapy can induce radioresistance in cancer cells, thereby resulting in a poor response rate. Our results demonstrated that pretreatment with ISO induced radiosensitizing effect in A549 cells using colony formation, micronucleus, and γH2AX foci assays. In addition, ISO pretreatment significantly enhanced the radiation-induced incidence of apoptosis, the collapse of mitochondrial membrane potential, and the expressions of proteins associated with cellular apoptosis and suppressed the upregulation of NF-κBp65 induced by irradiation in A549 cells. Interestingly, the expression of interleukin-13 (IL-13), an anti-inflammatory cytokine, was positively correlated with the ISO-mediated radiosensitization of A549 cells. The knockdown of IL-13 expression by RNA interference decreased the IL-13 level and thus reduced ISO-mediated radiosensitivity in cells. We also found that the IR-induced NF-κB signaling activation was inhibited by ISO pretreatment, and it was abrogated in IL-13 silenced cells. We speculated that ISO may confer radiosensitivity on A549 cells via increasing the expression of IL-13 and inhibiting the activation of NF-κB. To our knowledge, this is the first report demonstrating the effects of ISO treatment on the responsiveness of lung cancer cells to irradiation through IL-13 and the NF-κB signaling pathway. In summary, ISO is a naturally occurring radiosensitizer with a potential application in adjuvant radiotherapy.

15.
Biomacromolecules ; 9(1): 363-8, 2008 Jan.
Article in English | MEDLINE | ID: mdl-18163571

ABSTRACT

For imitating the active site of antioxidant selenoenzyme glutathione peroxidase (GPx), an artificial enzyme selenosubtilisin was employed as a scaffold for reconstructing substrate glutathione (GSH) specific binding sites by a bioimprinting strategy. GSH was first covalently linked to selenosubtilisin to form a covalent complex GSH-selenosubtilisin through a Se-S bond, then the GSH molecule was used as a template to cast a complementary binding site for substrate GSH recognition. The bioimprinting procedure consists of unfolding the conformation of selenosubtilisin and fixing the new conformation of the complex GSH-selenosubtilisin. Thus a new specificity for naturally occurring GPx substrate GSH was obtained. This bioimprinting procedure facilitates the catalytic selenium moiety of the imprinted selenosubtilisin to match the reactive thiol group of GSH in the GSH binding site, which contributes to acceleration of the intramolecular catalysis. These imprinted selenium-containing proteins exhibited remarkable rate enhancement for the reduction of H2O2 by GSH. The average GPx activity was found to be 462 U/micromol, and it was approximately 100 times that for unimprinted selenosubtilisin. Compared with ebselen, a well-known GPx mimic, an activity enhancement of 500-fold was observed. Detailed steady-state kinetic studies demonstrated that the novel selenoenzyme followed a ping-pong mechanism similar to the naturally occurring GPx.


Subject(s)
Glutathione Peroxidase/metabolism , Molecular Mimicry , Selenium/metabolism , Binding Sites , Catalysis , Electrophoresis, Polyacrylamide Gel , Substrate Specificity
16.
FEBS J ; 274(15): 3846-54, 2007 Aug.
Article in English | MEDLINE | ID: mdl-17617230

ABSTRACT

A 6A,6A'-dicyclohexylamine-6B,6B'-diselenide-bis-beta-cyclodextrin (6-CySeCD) was designed and synthesized to imitate the antioxidant enzyme glutathione peroxidase (GPX). In this novel GPX model, beta-cyclodextrin provided a hydrophobic environment for substrate binding within its cavity, and a cyclohexylamine group was incorporated into cyclodextrin in proximity to the catalytic selenium in order to increase the stability of the nucleophilic intermediate selenolate. 6-CySeCD exhibits better GPX activity than 6,6'-diselenide-bis-cyclodextrin (6-SeCD) and 2-phenyl-1,2-benzoisoselenazol-3(2H)-one (Ebselen) in the reduction of H(2)O(2), tert-butyl hydroperoxide and cumenyl hydroperoxide by glutathione, respectively. A ping-pong mechanism was observed in steady-state kinetic studies on 6-CySeCD-catalyzed reactions. The enzymatic properties showed that there are two major factors for improving the catalytic efficiency of GPX mimics. First, the substrate-binding site should match the size and shape of the substrate and second, incorporation of an imido-group increases the stability of selenolate in the catalytic cycle. More efficient antioxidant ability compared with 6-SeCD and Ebselen was also seen in the ferrous sulfate/ascorbate-induced mitochondria damage system, and this implies its prospective therapeutic application.


Subject(s)
Chlorine/chemistry , Cyclodextrins/chemistry , Cyclodextrins/metabolism , Glutathione Peroxidase/metabolism , Organoselenium Compounds/chemistry , Organoselenium Compounds/metabolism , Selenium/chemistry , beta-Cyclodextrins/chemistry , beta-Cyclodextrins/metabolism , Animals , Catalysis , Cattle , Cyclodextrins/chemical synthesis , Kinetics , Mitochondria, Heart/metabolism , Molecular Structure , Organoselenium Compounds/chemical synthesis , Oxidative Stress , beta-Cyclodextrins/chemical synthesis
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