ABSTRACT
Developing multifunctional metal-organic frameworks (MOFs) is a new research trend. MOFs have shown remarkable performances in both proton conduction and fluorescence sensing, but the MOFs integrating the two performances are scarce. Herein, a Co-MOF, [Co6(oba)4(Hatz)(atz)(H2O)2(µ3-OH)2(µ2-OH)]·H2O (1, H2oba = 4,4-oxybis(benzoic acid), Hatz = 5-amino-1H tetrazole), has been assembled by Co2+ ions with H2oba and Hatz ligands, providing a unique example of multifunctional MOFs with both proton conduction and fluorescence sensing performances. The framework of 1 displays a pillar-layer structure built by the oba ligand as a pillar and a layer composed of Co-clusters and atz linkers. Because large-scale single crystals of 1 were successfully synthesized, the proton conduction ability of 1 was investigated using single crystal samples. 1 exhibits highly anisotropic conduction with conductivity values of 1.1 × 10-3 S cm-1 along the [001] direction and 9.1 × 10-6 S cm-1 along the [010] direction at 55 °C and 95% RH, respectively. Meanwhile, the fluorescence sensing of 1 towards metal ions was studied in aqueous solutions. Attractively, 1 may sensitively and selectively detect Fe3+ ions in the presence of other interfering ions by fluorescence quenching.
ABSTRACT
An effective method to eliminate grain boundary resistance of crystalline vanadoborate electrolyte was developed. This method involved the addition of glycerol to result in the formation of many hydrogen bonds between crystal grains, facilitating a rapid transfer of protons across grain boundaries. Using this method, the intrinsic conduction of vanadoborate electrolyte was fully reflected in its bulk materials, valuable for advancing our understanding of vanadoborate electrolytes and for promoting the application of these electrolytes.
ABSTRACT
Through one-pot mechanochemical synthesis, a series of guests [imidazole, (aminomethyl)phosphonic acid, urea and sulfamic acid] are rapidly encapsulated into the pores of MOF NENU-3 while the MOF is formed. The synthesis of a MOF loaded with functional guests that used to take several days and require a multistep procedure can now be completed in one step within several minutes. The proton conductivities of the obtained composites increased by 2-3 orders of magnitude compared with NENU-3.
ABSTRACT
Effecting the synergistic function of single metal atom sites and their supports is of great importance to achieve high-performance catalysts. Herein, we successfully fabricate polyoxometalates (POMs)-stabilized atomically dispersed platinum sites by employing three-dimensional metal-organic frameworks (MOFs) as the finite spatial skeleton to govern the accessible quantity, spatial dispersion, and mobility of metal precursors around each POM unit. The isolated single platinum atoms (Pt1) are steadily anchored in the square-planar sites on the surface of monodispersed Keggin-type phosphomolybdic acid (PMo) in the cavities of various MOFs, including MIL-101, HKUST-1, and ZIF-67. In contrast, either the absence of POMs or MOFs yielded only platinum nanoparticles. Pt1-PMo@MIL-101 are seven times more active than the corresponding nanoparticles in the diboration of phenylacetylene, which can be attributed to the synergistic effect of the preconcentration of organic reaction substrates by porous MOFs skeleton and the decreased desorption energy of products on isolated Pt atom sites.
ABSTRACT
Kinesin-1 is a motor protein moving along a microtubule with its two identical motor heads dimerized by two neck linkers and a coiled-coil stalk. When both motor heads bind the microtubule, an internal strain is built up between the two heads, which is indispensable to ensure proper coordination of the two motor heads during kinesin-1's mechanochemical cycle. The internal strain forms a tensile force along the neck linker that tends to unwind the neck coiled coil (NCC). Experiments showed that the kinesin-1's NCC has a high antiunwinding ability compared with conventional coiled coils, which was mainly attributed to the enhanced hydrophobic pressure arising from the unconventional sequence of kinesin-1's NCC. However, hydrophobic pressure cannot provide the shearing force which is needed to balance the tensile force on the interface between two helices. To find out the true origin of the mechanical stability of kinesin-1's NCC, we perform a novel and detailed mechanical analysis for the system based on molecular dynamics simulation at an atomic level. We find that the needed shearing force is provided by a buckle structure formed by two tyrosines which form effective steric hindrance in the presence of tensile forces. The tensile force is balanced by the tensile direction component of the contact force between the two tyrosines which forms the shearing force. The hydrophobic pressure balances the other component of the contact force perpendicular to the tensile direction. The antiunwinding strength of NCC is defined by the maximum shearing force, which is finally determined by the hydrophobic pressure. Kinesin-1 uses residues with plane side chains, tryptophans and tyrosines, to form the hydrophobic center and to shorten the interhelix distance so that a high antiunwinding strength is obtained. The special design of NCC ensures exquisite cooperation of steric hindrance and hydrophobic pressure that results in the surprising mechanical stability of NCC.
Subject(s)
Kinesins/chemistry , Protein Stability , Computer Simulation , Hydrogen Bonding , Hydrophobic and Hydrophilic Interactions , Protein Domains , Protein Structure, SecondaryABSTRACT
A polyoxovanadate-based nickel-organic framework, [Ni(bib)2]{V2O6}({V6}-MOF, bib = 1,4-bis(1H-imidazoly-1-yl)benzene), was facilely prepared under gentle hydrothermal conditions and structurally characterized. Single-crystal X-ray diffraction analysis indicates that the {V6} cluster in the {V6}-MOF is constructed of two VO5 tetragonal pyramids and four VO4 tetrahedrons via the apex sharing of O atoms, presenting a hollow Linqvist-like structure, which is different from these reported hexanuclear vanadium clusters. The {V6}-MOF not only expands the structure of polyoxovanadates (POVs) but also catalyzes the rapid detoxification of mustard gas simulant (2-chloroethyl ethyl sulfide, CEES) at 25 °C. The catalytic results were determined by means of GC, GC-MS, and 1H NMR. Using {V6}-MOF as a heterogeneous catalyst, CEES underwent catalyzed oxidation to only nontoxic product 2-chloroethyl ethyl sulfoxide (CEESO) within 40 min, and the conversion and selectivity were almost 100%. In addition, {V6}-MOF exhibits high sustainability, and no obvious reductions in conversion and selectivity are observed after five runs.
ABSTRACT
AtAMT1;3 is a major contributor to high-affinity ammonium uptake in Arabidopsis roots. Using a stable electrophysiological recording strategy, we demonstrate in Xenopus laevis oocytes that AtAMT1;3 functions as a typical high-affinity NH4 + uniporter independent of protons and Ca2+. The findings that AtAMT1;3 transports methylammonium (MeA+, a chemical analog of NH4 +) with extremely low affinity (K m in the range of 2.9-6.1 mM) led to investigate the mechanisms underlying substrate binding. Homologous modeling and substrate docking analyses predicted that the deduced substrate binding motif of AtAMT1;3 facilitates the binding of NH4 + ions but loosely accommodates the binding of MeA+ to a more superficial location of the permeation pathway. Amongst point mutations tested based on this analysis, P181A resulted in both significantly increased current amplitudes and substrate binding affinity, whereas F178I led to opposite effects. Thus these 2 residues, which flank W179, a major structural component of the binding site, are also important determinants of AtAMT1;3 transport capacity by being involved in substrate binding. The Q365K mutation neighboring the histidine residue H378, which confines the substrate permeation tunnel, affected only the current amplitudes but not the binding affinities, providing evidence that Q365 mainly controls the substrate diffusion rate within the permeation pathway.
ABSTRACT
To elucidate the mechanism of bloom formation on fruit surface of cucumber, we investigated silicon absorption and bloom formation on fruit surface of cucumber with 'Shannong No. 5' (Cucumis sativus) as scion, 'Yunnan figleaf gourd' (Cucurbita ficifolia, weak de-blooming ability) and 'Huangchenggen No. 2' (C. moschata, strong de-blooming ability) as rootstocks in solar greenhouse at winter-spring and autumn-winter growing seasons. Experimental conditions inclu-ded: T1 with temperature 28 â/18 â (day/night), relative humidity 55%/65%, photosynthetic photon flux density 600 µmol·m-2·s-1, and T2 with temperature 22 â/12 â (day/night), relative humidity 85%/95%, photosynthetic photon flux density 300 µmol·m-2·s-1. We examined environmental effects on silicon absorption and expression of silicon transporter genes in. The amount of bloom on cucumber fruit surface at winter-spring growing season dramatically increased compared with autumn-winter season. 'Yunnan figleaf gourd' grafted cucumber was more heavily affected by the cultivation season than self-rooted and 'Huangchenggen No. 2' grafted cucumber. In the same cultivation season, 'Yunnan figleaf gourd' grafted cucumber had the highest amount of bloom on fruit surface and silicon content, while own-rooted and 'Huangchenggen No. 2' grafted cucumber had the medium and least amount of bloom and silicon content. Silicon content in each organ and expression of silicon transporter genes in cucumber leaves and roots under T1 environment were significantly increased compared with T2. 'Yunnan figleaf gourd' grafted cucumber had the highest contents of silicon in each organ and expression of silicon transporter genes in leaves in the same environment, followed by own-rooted and 'Huangchenggen No. 2' grafted cucumber. In conclusion, environmental conditions affect absorption and allocation of silicon in cucumber plants, with conseuqnece on bloom formation on fruit surface. Suitable environmental conditions, including temperature, humidity and light, are beneficial to reduce the bloom formation on cucumber fruit surface. High temperature, strong light, and low humidity will increase bloom amount on cucumber fruit. Rootstocks have significant effects on silicon absorption and fruit bloom formation of grafted cucumber.
Subject(s)
Cucumis sativus , China , Fruit , Plant Roots , SiliconABSTRACT
AIM: To evaluate whether recombinant complement factor B (CFB) short hairpin RNA (shRNA) reduces laser-induced choroidal neovascularization (CNV) in rats. METHODS: Laser-induced rat CNV model was established, and then the animals underwent fundus fluorescence angiography (FFA) and hematoxylin and eosin (HE) staining. On day 3 and 7 after photocoagulation, the expression of CFB and membrane attack complex (MAC) was detected by immunhischemistry. A recombinant CFB-shRNA plasmid was constructed. CFB and scrambled shRNA plasmids were intravenous injected into rats via the tail vein on the day of laser treatment, respectively. On day 7, the incidence of CNV was determined by FFA, and the expression of CFB and vascular endothelial growth factor (VEGF) in retinal pigment epithelium (RPE)/choroidal tissues was detected by immunhischemistry, Western blot and/or semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) in CFB and scrambled shRNA groups. The possible adverse effects of CFB-shRNA injection were assessed by transmission electron microscopy and electroretinography. RESULTS: FFA and HE results indicated that a laser-induced rat CNV model was successfully established on day 7 after photocoagulation. The expression of CFB and MAC was extremely weak in normal retina and choroid, and increased on day 3 after photocoagulation. However, it started to reduce on day 7. CFB shRNA plasmid was successfully constructed and induced CFB knockdown in the retinal and choroidal tissues. FFA showed CFB knockdown significantly inhibited incidence of CNV in rats. Moreover, CFB knockdown significantly inhibited the expression of VEGF in RPE/choroidal tissues. CFB shRNA caused no obvious side effects in eyes. CONCLUSION: CFB knockdown significantly inhibits the formation and development of CNV in vivo through reducing the expression of VEGF, which is a potential therapy target. The alternative pathway of complement activation plays an important role in CNV formation.
ABSTRACT
A novel method is proposed to fabricate polyoxometalate-based metal-organic frameworks (POM@MOFs) by using the ability of POMs to oxidize metals to cations. The morphology-controlled growth of POM@MOFs (NENU-n) is achieved in the absence of any modulators and an effect of polyoxoanion charge number on morphology is observed.
ABSTRACT
A pure-inorganic framework based on {P4MoV4MoVI2} clusters with rich phosphate groups has been synthesized. It underwent a single crystal to single crystal conversion in air to form a new framework with changes both in the metal valent state and coordination environment. The new framework exhibits an ultra-high proton conductivity of 1.33 × 10-2 S cm-1 at 95 °C and 98% relative humidity and excellent stability.
ABSTRACT
BACKGROUND: Accumulating evidence shows that circular RNAs (circRNAs) plays vital roles in tumor progression. However, the biological functions of circRNAs in laryngeal squamous cell carcinoma (LSCC) metastasis is still unclear. METHODS: qRT-PCR was used to detect circFLNA, miRNAs and FLNA mRNA expression. Transwell assay and western blot were performed to evaluate cell migration ability and to detect FLNA, MMP2 and MLK1 protein expression, respectively. RNA pull-down analysis was used to find the binding-miRNAs of circFLNA. Luciferase reporter assay was used to examine the effect of circFLNA on miRNAs and miR-486-3p on FLNA expression. RESULTS: In this study, we confirmed that a Filamin A (FLNA)-derived hsa_circ_0092012 known as circFLNA, was upregulated in LSCC, and the higher expression of circFLNA was correlated with LSCC lymph node metastasis. Increased circFLNA facilitates LSCC cell migration ability through upregulating FLNA and MMP2 protein expression. Mechanistically, we find that circFLNA sponges miR-486-3p in LSCC cells, relieving miR-486-3p-induced repression of FLNA which promotes LSCC cell migration. Accordingly, FLNA mRNA is overexpressed in LSCC tissues and a higher FLNA level is correlated with poor survival. Dysregulation of the circFLNA/miR-486-3p/FLNA regulatory pathway contributes to LSCC migration. CONCLUSIONS: In summary, our study sheds light on the regulatory mechanism of circFLNA in LSCC migration via sponging miR-486-3p, which downregulates the FLNA protein expression. Targeting circFLNA/miR-486-3p/FLAN axis provides a potential therapeutic target for aggressive LSCC.
ABSTRACT
A novel bacterial strain A7.6T was isolated from the sediments collected near the Zhairuo Island located in the East China Sea and characterized using a polyphasic approach. Cells were Gram-stain-negative, rod-shaped, non-spore forming, non-flagellated but motile by gliding. The strain was aerobic, positive for oxidase and catalase activities. The strain can grow at 4-35 °C, pH 5.5-9.0, and 0-3% (w/v) NaCl concentration. The major polar lipid was phosphatidylethanolamine, the predominant fatty acids (> 10%) were iso-C15:0 and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c). The genomic G+C content was 33.6 mol% and the major respiratory quinone was menaquinone 6. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain A7.6T belonged to the genus Flavobacterium and was closely related to Flavobacterium tistrianum GB 56.1T (98.4% similarity), F. nitrogenifigens NXU-44T (98.4%), F. ginsenosidimutans THG 01T (98.0%) and F. anhuiense D3T (97.7%). Average nucleotide identities and digital DNA-DNA hybridizations values for genomes ranged from 75.9 to 91.4% and 21.4 to 43.9% between strain A7.6T and its closest phylogenetic neighbors. The polyphasic characterization indicated that strain A7.6T represented a novel species of the genus Flavobacterium, for which the name Flavobacterium sharifuzzamanii is proposed. The type strain is A7.6T (= KCTC 62405T = MCCC 1K03485T). The NCBI GenBank accession number for the 16S rRNA gene of A7.6T is MH396692, and for the genome sequence is QJGZ00000000. The digital protologue database (DPD) Taxon Number is TA00643.
Subject(s)
Flavobacterium/classification , Flavobacterium/physiology , Geologic Sediments/microbiology , Oceans and Seas , Phylogeny , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/analysis , Flavobacterium/chemistry , Genome, Bacterial/genetics , Hydrogen-Ion Concentration , Phospholipids/analysis , Quinones/analysis , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Sodium Chloride , TemperatureABSTRACT
In this paper, a sandwich type of polyoxophosphotungstate K7Na3[Cu4(H2O)2(PW9O34)2]·20H2O (abbr. Cu4(PW9)2) was selected as an active unit to construct inorganic/organic hybrid materials. The ordered honeycomb structure of the surfactant-encapsulated polyoxometalate (SEP) complex was successfully obtained by using a famous breath figure method. Cu4(PW9)2 maintains typical features as confirmed by IR spectra, magnetization hysteresis studies and UV-vis spectra. An investigation of the effects of the relative humidity on the morphology of the honeycomb structures demonstrated a wide generality and high reproducibility of the formation of self-organized honeycomb-patterned films. In the present experiment, this magnetically ordered honeycomb was successfully deposited on indium tin oxide (ITO) substrates by directly casting the SEP chloroform solution. The cyclic voltammetric responses showed the redox couples of Cu4(PW9)2. The electrochemical behavior of the electrode was studied. In addition, the modified electrode exhibited high electrocatalytic activity for the reduction of BrO3-, which offered a new POM-based film material for the electrochemical fields.
ABSTRACT
The objective of this study was to examine the role and possible mechanisms of toll-like receptor 2 (TLR2) in high-mobility group box chromosomal protein 1 (HMGB1)-induced mouse mesangial cell (MMC) proliferation and glomeruli proliferation of MRL/Fas(lpr) mice. First, the expression of proliferating cell nuclear antigen (PCNA), TLR2 and Forkhead box protein O1 (FoxO1) messenger RNA (mRNA) and protein in the glomeruli of MRL/Fas(lpr) mice was quantified, and the correlation with cell proliferation of glomeruli was analyzed. Then, lipopolysaccharide (LPS), TLR2 neutralization antibody, and small hairpin TLR2 (shTLR2) were used to confirm the role of TLR2 in HMGB1-induced MMC proliferation. Furthermore, wild-type FoxO1 (WT-FoxO1) vector was used to investigate the effect of FoxO1 pathway on HMGB1-induced MMC proliferation. Finally, electroporation was used to knockdown TLR2 in the glomeruli of MRL/Fas(lpr) mice, and renal function, FoxO1, and PCNA expression were detected. The results showed that the TLR2 expression was upregulated and FoxO1 expression was decreased in the glomeruli of MRL/Fas(lpr) mice, and these effects were significantly correlated with cell proliferation of the glomeruli. In vitro, the TLR2 neutralization antibody and the WT-FoxO1 vector, both reduced the MMC proliferation levels induced by HMGB1. The TLR2 neutralization antibody also blocked the HMGB1-dependent activation of the FoxO1 pathway and cell proliferation. In addition, transfection with shTLR2 decreased the proliferation levels and PCNA expression induced by HMGB1. In vivo, treatment with shTLR2 significantly reduced the PCNA expression in the glomeruli of MRL/Fas(lpr) mice and improved renal function. In addition, treatment with shTLR2 or blocking of TLR2 also reduced the translocation of FoxO1. Thus, TLR2 plays a critical role in HMGB1-induced glomeruli cell proliferation through the FoxO1 signaling pathway in lupus nephritis.
Subject(s)
Forkhead Box Protein O1/metabolism , Lupus Nephritis/immunology , Toll-Like Receptor 2/metabolism , Animals , Antibodies, Blocking/immunology , Cell Proliferation , Cells, Cultured , Disease Models, Animal , Female , Forkhead Box Protein O1/genetics , HMGB1 Protein/physiology , Humans , Mice , Mice, Mutant Strains , Proliferating Cell Nuclear Antigen/genetics , Proliferating Cell Nuclear Antigen/metabolism , RNA, Small Interfering/genetics , Signal Transduction/immunology , Toll-Like Receptor 2/genetics , Toll-Like Receptor 2/immunology , fas Receptor/geneticsABSTRACT
Although selective COX-2 inhibitors have cancer-preventive effects and induce apoptosis, the mechanisms underlying these effects are not fully understood. This study investigated the effects of nimesulide, a selective COX-2 inhibitor, on apoptosis and on the JAK/STAT signaling pathway in Eca-109 human esophageal squamous carcinoma cells. The effects and mechanisms of nimesulide on Eca-109 cell growth were studied in culture and in nude mice with Eca-109 xenografts. Cells were cultured with or without nimesulide and/or the JAK2 inhibitor AG490. Cell proliferation was evaluated using the MTT assay, and apoptosis was investigated. COX-2 mRNA expression was measured using reverse transcription polymerase chain reaction, and protein expression was detected by Western blot analysis, immunohistochemistry, and flow cytometry. Nimesulide significantly inhibited Eca-109 cell viability in vitro in a dose- and time-dependent manner (P<0.05). Nimesulide also induced apoptosis, which was accompanied by a significant decrease in the expression of COX-2 and survivin and an increase in caspase-3 expression. Nimesulide downregulated the phosphorylation levels of JAK2 and STAT3, and JAK2 inhibition by AG490 significantly augmented both nimesulide-induced apoptosis and the downregulation of COX-2 and survivin (P<0.05). In vivo, nimesulide inhibited the growth of Eca-109 tumors and the expression of p-JAK2 and p-STAT3. Thus, nimesulide downregulates COX-2 and survivin expression and upregulates caspase-3 expression in Eca-109 cells, by inactivating the JAK2/STAT3 pathway. These effects may mediate nimesulide-induced apoptosis and growth inhibition in Eca-109 cells in vitro and in vivo.
Subject(s)
Carcinoma, Squamous Cell/metabolism , Cyclooxygenase 2/metabolism , Esophageal Neoplasms/metabolism , Janus Kinase 2/metabolism , STAT3 Transcription Factor/metabolism , Sulfonamides/pharmacology , Animals , Apoptosis/drug effects , Carcinoma, Squamous Cell/drug therapy , Cell Line, Tumor , Cell Proliferation/drug effects , Esophageal Neoplasms/drug therapy , Esophageal Squamous Cell Carcinoma , Female , Humans , Inhibitor of Apoptosis Proteins/genetics , Mice, Inbred BALB C , Mice, Nude , Signal Transduction/drug effectsABSTRACT
Our previous experiment confirmed that high-mobility group box chromosomal protein 1 (HMGB1) was involved in the pathogenesis of Lupus nephritis (LN) by upregulating the proliferation of the mouse mesangial cell line (MMC) through the cyclin D1/CDK4/p16 system, but the precise mechanism is still unknown. Therefore, in the present study, we demonstrated that HMGB1 induced the proliferation of MMC cells in a time- and concentration-dependent manner, downregulated phosphatase and tensin homolog deleted on chromosome ten (PTEN) expression, increased the level of Akt serine 473 phosphorylation, and induced p65 subunit nuclear translocation. The overexpression of PTEN prevented the upregulation of HMGB1-induced proliferation by blocking the activation of Akt. The knockdown of Akt by siRNA technology and blocking the nuclear factor-κB (NF-κB) pathway using pyrrolidine dithiocarbamate (PDTC) and SN50, inhibitors of NF-κB, both attenuated the HMGB1-induced proliferation by counteracting the activation of the cyclin D1. In addition, while sh-Akt partly blocked the nuclear translocation of the p65 subunit, PDTC did not affect the activation of the Akt induced by HMGB1 in MMC cells. These findings indicate that HMGB1 induced the proliferation of MMC cells by activating the PTEN/phosphoinositide-3-kinase (PI3K)/Akt/NF-κB signaling pathway.
Subject(s)
Cyclin D1/genetics , HMGB1 Protein/genetics , Lupus Nephritis/genetics , Mesangial Cells/metabolism , PTEN Phosphohydrolase/genetics , Animals , Cell Proliferation , Cyclin D1/metabolism , Gene Knockdown Techniques , HMGB1 Protein/metabolism , Lupus Nephritis/metabolism , Lupus Nephritis/pathology , Mice , NF-kappa B/genetics , NF-kappa B/metabolism , Oncogene Protein v-akt/genetics , Oncogene Protein v-akt/metabolism , PTEN Phosphohydrolase/metabolism , Phosphatidylinositol 3-Kinases/genetics , Phosphorylation , Signal Transduction/geneticsABSTRACT
Conventional kinesin's force generation process always takes place on the leading head and the generated force is transmitted to the trailing head through two neck linkers. To guarantee a strong force to be transmitted to the trailing head so that it can be detached from microtubule surface, the neck linker of the leading head must have a large enough forward displacement, which is proposed to be achieved by the amplifying function of the ß-domain. However, the experimental result shows that the forward displacement of the ß-domain itself appears too small. To elucidate the function of the ß-domain, we make a detailed analysis of the mechanical relationship between the two motor heads and, based on the results of molecular dynamics simulation and mechanical analysis, we calculate the forward displacement of the neck linker of the leading head during the ATP binding induced motor head rotation. We show that ß-domain achieves its amplifying function together with ß0, so that neck linker can have a forward displacement during motor head rotation. This displacement of neck linker is large enough to cause detachment of the trailing head. Based on these results, a possible initiation mechanism of neck linker docking is proposed.
Subject(s)
Kinesins/chemistry , Kinesins/metabolism , Mechanical Phenomena , Adenosine Triphosphate/metabolism , Molecular Dynamics Simulation , Protein Structure, Tertiary , RotationABSTRACT
Podocyte apoptosis contributes to the pathogenesis of diabetic nephropathy (DN). However, the mechanisms that mediate high glucose (HG)-induced podocyte apoptosis remain poorly understood. Conditionally immortalized mouse podocytes were cultured in HG medium. A chemical inhibitor or a specific short-hairpin RNA (shRNA) vector was used to inhibit the activation of the Notch pathway and the PI3K/Akt pathway in HG-treated podocytes. Western blotting and real-time PCR were used to evaluate the levels of Notch, PI3K/Akt, and apoptotic pathway signaling. The apoptosis rate of HG-treated podocytes was assessed by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling and annexin V/propidium iodide staining. In HG-treated podocytes, PI3K/Akt pathway activation prevented podocyte apoptosis in the early stage of HG stimulation and Notch pathway-induced podocyte apoptosis in the late stage of HG stimulation. The inhibition of the Notch pathway or the activation of the PI3K/Akt pathway prevented cell apoptosis in HG-treated podocytes. These findings suggest that the Notch and PI3K/Akt pathways may mediate HG-induced podocyte apoptosis.
Subject(s)
Apoptosis/drug effects , Apoptosis/physiology , Glucose/pharmacology , Oncogene Protein v-akt/physiology , Phosphatidylinositol 3-Kinases/physiology , Podocytes/drug effects , Podocytes/physiology , Receptors, Notch/physiology , Signal Transduction/physiology , Animals , Annexin A5 , Apoptosis/genetics , Apoptosis Regulatory Proteins/genetics , Apoptosis Regulatory Proteins/physiology , Blotting, Western , Cells, Cultured , Chromones/pharmacology , Enzyme Inhibitors/pharmacology , Genetic Vectors , In Situ Nick-End Labeling , Mice , Morpholines/pharmacology , Oligopeptides/pharmacology , RNA, Small Interfering/genetics , Real-Time Polymerase Chain Reaction , Receptor, Notch1/metabolism , Receptor, Notch1/physiology , Up-Regulation/drug effectsABSTRACT
OBJECTIVE: To explore the possible mechanism of lipid deposition induced by interferon-gamma (IFN-gamma). METHODS: The mouse mesangial cells (MMC) were randomly divided into control group, stimulation group, stimulation + control vector group (sh-HMGB1) and stimulation+ specific sh-vector group (sh-SREBP-1). RT-PCR was used to detect the expression of HMGB1, SREBP-1 and fatty acid synthetase (FAS) mRNA; the protein expression was determined by Western blot. RESULTS: The Oil Red O staining revealed that the mouse mesangial cells showed significant lipid droplet in IFN-gamma group. IFN-gamma up-regulated the expression of HMGB1, SREBP-1, FAS mRNA and protein time-dependently; Transfection of MMC with HMGB1 siRNA resulted in the suppression of SREBP-1, FAS protein levels induced by IFN-gamma, following with decrease of lipid deposition. Stimulation with HMGB1 markedly induced expression of SREBP-1, FAS expression and peaked at 8 h, decreased at 12 h compared with that at 8 h. Sh-SREBP-1 decreased the lipid deposition induced by HMGB1 in MMC. CONCLUSION: IFN-gamma might induce lipid deposition in mouse mesangial cells partly by up-regulating the expression of HMGB1/SREBP-1/FAS.