Song, C-G*
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Macranthoside B (MB) is a triterpenoid saponin extracted from Lonicera macranthoides, a traditional Chinese medicine. In the current study, we investigated the anticancer potential of MB in various cancer cells and elucidated its underlying mechanisms. MB exposure inhibited cell proliferation, induced mitochondrial membrane potential (MMP) loss, increased sub-G1 accumulation, and resulted in cleavage of caspase-3 and PARP, which are reflective of apoptosis. In HeLa cells, MB induced down-regulation of SOD2 and GPx1, phosphorylation of Akt and PDK1, and thus promoted ROS-mediated apoptosis. This was further supported by the protection of sub-G1 accumulation, MMP loss, cleavage of caspase-3 and PARP in the presence of N-acetylcysteine (NAC). Additionally, MB induced cell death via down-regulation of ubiquitin-like with PHD and ringfinger domains 1 (UHRF1) and Bcl-xL. Taken together, this study provides a new insight into the apoptosis- inducing potential of MB, and its molecular mechanisms are associated with an increase in oxidative stress and inhibition of the PDK1/Akt pathway.
- MeSH
- adenokarcinom * MeSH
- apoptóza MeSH
- HeLa buňky MeSH
- kaspasa 3 metabolismus MeSH
- lidé MeSH
- membránový potenciál mitochondrií MeSH
- nádorové buněčné linie MeSH
- PARP inhibitory farmakologie MeSH
- proteiny vázající zesilovač transkripce CCAAT metabolismus farmakologie MeSH
- protoonkogenní proteiny c-akt metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- saponiny * farmakologie MeSH
- ubikvitinligasy metabolismus farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Cysteine protease is a superfamily of widespread proteolytic enzymes and plays a major role in larval invasion, migration, exsheathing, survival and immune evasion in parasites. In the present study, the gene coding cysteine proteinase of the nematode Trichinella spiralis (Owen, 1835) was cloned into pQE-80L and subsequently expressed in E. coli JM109. The rTsCP was purified and its antigenicity was identified by Western blot and ELISA. Using anti-rTsCP serum the native TsCP was identified in muscle larval crude proteins. The results of quantitative real-time PCR and immunofluorescence test demonstrated that the TsCP was expressed in all stages of T. spiralis and located mainly in cuticle, stichosome and reproductive organs. The immunisation of mice with rTsCP elicited Th2-predominant immune responses. Anti-rTsCP antibodies could partially inhibit the in vitro larval invasion of intestinal epithelial cells and kill the newborn larvae by an antibody-dependent cell-mediated dose-dependent cytotoxicity. The vaccinated mice exhibited a 54% reduction of adults and a 33% reduction of muscle larvae following challenge infection. The results suggested that the TsCP might be an indispensable protein in Trichinella invasion, development and survival of T. spiralis in hosts, and could be a potential vaccine target against infection.
- MeSH
- cysteinové proteasy genetika metabolismus MeSH
- Escherichia coli genetika metabolismus MeSH
- exprese genu MeSH
- geneticky modifikované mikroorganismy genetika MeSH
- klonování DNA MeSH
- larva enzymologie genetika růst a vývoj MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- organismy bez specifických patogenů MeSH
- proteiny červů genetika metabolismus MeSH
- sekvenční analýza DNA veterinární MeSH
- Trichinella spiralis enzymologie genetika růst a vývoj MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Enterovirus 71 (EV71) infection and its associated hand-foot-mouth disease is a significant public health problem. The purpose of this study is to develop a novel vaccine to prevent EV71 infection. Bacillus subtilis spores were engineered to express VP1 protein of EV71 with CotB as carrier protein. The recombination was tested in adult mice for the ability to induce immune responses. Mice were inoculated orally and intranasally simultaneously with the spores. The vaccine’s efficacy on stimulating immune responses was evaluated by measuring the titer of anti-VP1 IgG and IgA with enzyme-linked-immunosorbent serologic assay (ELISA), and the number of VP1-specific T cells by ELIS-POT. Serum titers of IgG (0.41±0.05 vs 0.20±0.07) and IgA (0.24±0.02 vs 0.11±0.01) in mice immunized with recombinant CotB-VP1 spores were higher than that of mice immunized with nonrecombinant spores 1A771. Splenocytes from the group of mice receiving VP1 spores vaccination contained 1.69±0.52/10 4 VP1-specific T cells, which was greater than the 0.06±0.06/10 4 cells from the group of mice receiving nonrecombinant spores vaccination. In conclusion, B. subtilis spores displaying VP1 of EV71 are effective in stimulating cellular immunity and humoral immunity in mice.
- MeSH
- aktivní imunita imunologie účinky léků MeSH
- Bacillus subtilis * imunologie virologie MeSH
- bakteriální proteiny imunologie MeSH
- buněčná imunita imunologie účinky léků MeSH
- dospělí MeSH
- ELISA MeSH
- enterovirové infekce * prevence a kontrola MeSH
- enterovirus A lidský imunologie MeSH
- fluorescenční mikroskopie MeSH
- imunitní systém účinky léků MeSH
- imunoglobulin A imunologie MeSH
- imunoglobulin G imunologie MeSH
- imunologická odpověď na dávku MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- rekombinantní proteiny imunologie MeSH
- spory bakteriální imunologie virologie MeSH
- virové vakcíny * aplikace a dávkování chemická syntéza terapeutické užití MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- validační studie MeSH
- MeSH
- hypertriglyceridemie komplikace krev MeSH
- hypotyreóza epidemiologie etiologie krev MeSH
- lidé středního věku MeSH
- lidé MeSH
- prevalence MeSH
- rizikové faktory MeSH
- senioři MeSH
- studie případů a kontrol MeSH
- stupeň závažnosti nemoci MeSH
- triglyceridy krev MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- souhrny MeSH
To investigate the exact effects of dietary choline on hypertensive heart disease (HHD) and explore the potential mechanisms, male spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) were randomly divided into five groups as follows: WKY group, WKY + Choline group, SHR group, SHR + Choline group, and SHR + Choline + NaHS group. In choline treatment groups, rats were fed with 1.3% (w/v) choline in the drinking water for 3 months. The rats in the SHR + Choline + NaHS group were intraperitoneally injected with NaHS (100 micromol/kg/day, a hydrogen sulfide (H2S) donor) for 3 months. After 3 months, left ventricular ejection fraction (LVEF) and fractional shortening (LVFS), the indicators of cardiac function measured by echocardiography, were increased significantly in SHR as compared to WKY, although there was no significant difference in collagen volumes and Bax/Bcl-2 ratio between the two groups, indicating the early stage of cardiac hypertrophy. There was a significant decrease in LVEF and LVFS and an increase in collagen volumes and Bax/Bcl-2 ratio in SHR fed with choline, meanwhile, plasma H2S levels were significantly decreased significantly in SHR fed with choline accompanying by the decrease of cystathionine-gamma-lyase (CSE) activity. Three months of NaHS significantly increased plasma H2S levels, ameliorated cardiac dysfunction and inhibited cardiac fibrosis and apoptosis in SHR fed with choline. In conclusion, choline aggravated cardiac dysfunction in HHD through inhibiting the production of endogenous H2S, which was reversed by supplementation of exogenous H2S donor.
- MeSH
- funkce levé komory srdeční MeSH
- hypertenze * chemicky indukované MeSH
- kolagen MeSH
- krysa rodu rattus MeSH
- nemoci srdce * MeSH
- potkani inbrední SHR MeSH
- potkani inbrední WKY MeSH
- protein X asociovaný s bcl-2 MeSH
- sulfan * MeSH
- sulfidy * MeSH
- tepový objem MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Some metallo-supramolecular helical assemblies with size, shape, charge and amphipathic architectures similar to short cationic α-helical peptides have been shown to target and stabilise DNA G-quadruplexes (G4s) in vitro and downregulate the expression of G4-regulated genes in human cells. To expand the library of metallohelical structures that can act as efficient DNA G4 binders and downregulate genes containing G4-forming sequences in their promoter regions, we investigated the interaction of the two enantiomeric pairs of asymmetric Fe(II) triplex metallohelices with a series of five different DNA G4s formed by the human telomeric sequence (hTelo) and in the promoter regions of c-MYC, c-KIT, and k-RAS oncogenes. The metallohelices display preferential binding to G4s over duplex DNA in all investigated G4-forming sequences and induced arrest of DNA polymerase on template strands containing G4-forming sequences. Moreover, the investigated metallohelices suppressed the expression of c-MYC and k-RAS genes at mRNA and protein levels in HCT116 human cancer cells, as revealed by RT-qPCR analysis and western blotting.
Explaining species geographic distributions by macroclimate variables is the most common approach for getting mechanistic insights into large-scale diversity patterns and range shifts. However, species' traits influencing biophysical processes can produce a large decoupling from ambient air temperature, which can seriously undermine biogeographical inference. We combined stable oxygen isotope theory with a trait-based approach to assess leaf temperature during carbon assimilation (TL ) and its departure (ΔT) from daytime free air temperature during the growing season (Tgs ) for 158 plant species occurring from 3,400 to 6,150 m a.s.l. in Western Himalayas. We uncovered a general extent of temperature decoupling in the region. The interspecific variation in ΔT was best explained by the combination of plant height and δ13 C, and leaf dry matter content partly captured the variation in TL . The combination of TL and ΔT, with ΔT contributing most, explained the interspecific difference in elevational distributions. Stable oxygen isotope theory appears promising for investigating how plants perceive temperatures, a pivotal information to species biogeographic distributions.
- MeSH
- izotopy kyslíku MeSH
- listy rostlin * MeSH
- roční období MeSH
- teplota MeSH
- uhlík * MeSH
- Publikační typ
- časopisecké články MeSH
PGC1α is a transcriptional coactivator that plays key roles in mitochondrial biogenesis, so exploring its molecular characterization contributes to the understanding of mitochondrial function in cultured fish. In the present study, a full-length cDNA coding PGC1α was cloned from the liver of blunt snout bream (Megalobrama amblycephala) which covered 3741 bp with an open reading frame of 2646 bp encoding 881 amino acids. Sequence alignment and phylogenetic analysis revealed high conservation with other fish species, as well as other higher vertebrates. Comparison of the derived amino acid sequences indicates that, as with other fish, there is a proline at position 176 (RIRP) compared to a Thr in the mammalian sequences (RIRT). To investigate PGC1α function, three in vitro tests were carried out using primary hepatocytes of blunt snout bream. The effect of AMPK activity on the expression of PGC1α was determined by the culture of the hepatocytes with an activator (Metformin) or inhibitor (Compound C) of AMPK. Neither AMPK activation nor inhibition altered PGC1α expression. Knockdown of PGC1α expression in hepatocytes using small interfering RNA (si-RNA) was used to determine the role of PGC1α in mitochondrial biogenesis. No significant differences in the expression of NRF1 and TFAM, and mtDNA copy number were found between control and si-RNA groups. Also, hepatocytes were cultured with oleic acid, and the findings showed the significant reduction of mtDNA copy number in oleic acid group compared to control. Moreover, oleic acid down-regulated the expression of NRF1 and TFAM genes, while PGC1α expression remained unchanged. Our findings support the proposal that PGC1α may not play a role in mitochondrial biogenesis in blunt snout bream hepatocytes.
- Publikační typ
- časopisecké články MeSH
This study aimed to isolate and characterize a novel cellulolytic enzyme from black goat rumen by using a culture-independent approach. A metagenomic fosmid library was constructed from black goat rumen contents and screened for a novel cellulase. The KG37 gene encoding a protein of 858 amino acid residues (92.7 kDa) was isolated. The deduced protein contained a glycosyl hydrolase family 74 (GH74) domain and showed 77% sequence identity to two endo-1,4-β-glucanases from Fibrobacter succinogenes. The novel GH74 cellulase gene was overexpressed in Escherichia coli, and its protein product was functionally characterized. The recombinant GH74 cellulase showed a broad substrate spectrum. The enzyme exhibited its optimum activity at pH 5.0 and temperature range of 20-50 °C. The enzyme was thermally stable at pH 5.0 and at a temperature of 20-40 °C. The novel GH74 cellulase can be practically exploited to convert lignocellulosic biomass to value-added products in various industrial applications in future.
- MeSH
- bachor mikrobiologie MeSH
- celulasa chemie genetika izolace a purifikace MeSH
- Escherichia coli genetika metabolismus MeSH
- exprese genu MeSH
- Fibrobacter enzymologie genetika MeSH
- genetické testování MeSH
- genová knihovna MeSH
- klonování DNA MeSH
- koncentrace vodíkových iontů MeSH
- kozy mikrobiologie MeSH
- metagenom * MeSH
- metagenomika MeSH
- molekulová hmotnost MeSH
- rekombinantní proteiny genetika izolace a purifikace metabolismus MeSH
- sekvenční homologie MeSH
- stabilita enzymů MeSH
- substrátová specifita MeSH
- teplota MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
