JOURNAL OF CELLULAR AND MOLECULAR MEDICINE
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2021
·Vol. 25
·期 13
·6082-6093
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IF 5.31
Pin Wang,Juan Zhao,Xiulian Sun
Myocyte enhancer factor 2D (MEF2D) is predominantly expressed in the nucleus and associated with cell growth, differentiation, survival and apoptosis. Previous studies verified that phosphorylation at different amino acids determined MEF2's transcriptional activity which was essential in regulating downstream target genes expression. What regulates phosphorylation of MEF2D and affects its function has not been fully elucidated. Here, we uncovered that dual-specificity tyrosine phosphorylation regulated kinase 1A (DYRK1A), a kinase critical in Down's syndrome pathogenesis, directly bound to and phosphorylated MEF2D at Ser251 in vitro. Phosphorylation of MEF2D by DYRK1A significantly increased MEF2D protein level but attenuated its transcriptional activity, which resulted in decreased transcriptions of MEF2D target genes. Phosphorylation mutated Ser251A MEF2D exhibited enhanced transcriptional activity compared with wild type MEF2D. MEF2D and DYRK1A were observed co-localized in HEK293 and U87MG cells. Moreover, DYRK1A-mediated MEF2D phosphorylation in vitro might influence its nuclear export upon subcellular fractionation, which partially explained the reduction of MEF2D transcriptional activity by DYRK1A. Our results indicated that DYRK1A might be a regulator of MEF2D transcriptional activity and indirectly get involved in regulation of MEF2D target genes.
研究领域:
分子生物学
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细胞生物学
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神经生物学
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DOI
PubMed
高影响因子
DRUG METABOLISM AND DISPOSITION
·
2018
·
IF 3.64
Danyi Lu,Dong Dong,Qian Xie,Zhijie Li,Baojian Wu
UDP-glucuronosyltransferases (UGTs) play an important role in metabolism and detoxification of amine-containing chemicals. However, the disposition mechanisms for amines via UGT metabolism are not fully clear. Here we aim to investigate a potential role of UGT2B10 in N-glucosidation, and to determine the transporters for excretion of N-glucosides. We first established a HEK293 cell line stably overexpressing UGT2B10. Incubation of mianserin or cyclizine with the cells generated one N-glucuronide and one N-glucoside. Chemical inhibition (using specific chemical inhibitor Ko143) and biological inhibition (using specific shRNA) of BCRP resulted in a significant reduction in efflux of N-glucuronide. Similar results were observed when MRP4 was inhibited. Furthermore, inhibition of BCRP led to increased intracellular N-glucoside, whereas inhibition of MRP4 caused no changes in disposition of N-glucosides. The data overall indicated that BCRP not MRP4 was responsible for excretion of N-glucosides, whereas both BCRP and MRP4 contributed to excretion of N-glucuronides. Interestingly, down-regulation of N-glucuronidation led to increased N-glucosidation in the cells, supporting the glucosidation as a potential complementary pathway for conventional glucuronidation. In conclusion, UGT2B10 was for the first time identified as a N-glucosidation enzyme. Generated N-glucosides were excreted primarily by the BCRP transporter.
研究领域:
药理学
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生物化学
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分子生物学
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DOI
PubMed
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
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2015
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IF 4.152
Hui Wang,Haifeng Wei,Quan Huang,Hongmei Liu,Jingjing Hu,Yiyun Cheng,Jianru Xiao
Cationic dendrimers are associated with relatively low transfection efficacy and high toxicity in gene delivery. Surface modification of these dendrimers with functional ligands may resolve these issues. Here, we proposed a novel strategy to prepare surface-engineered dendrimers with high transfection efficacy and low toxicity on transfected cells. Several nucleobase analogues were modified on cationic dendrimers by a facile method. These nucleobase-modified dendrimers show improved transfection efficacy and reduced cytotoxicity compared to unmodified dendrimers on HEK293 and HeLa cells. Efficacy of the most efficient polymer is comparable to that of commercial transfection reagents such as SuperFect, PolyFect, and Lipofectamine 2000. The improved transfection efficacy of dendrimers after nucleobase modification is probably attributed to easier intracellular DNA unpacking and lower cytotoxicity. The results provide a valuable insight to guide the design of efficient and low cytotoxic gene vectors.
研究领域:
分子生物学
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基因治疗
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药剂学
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产品关联
DOI
PubMed
高影响因子
ACTA PHARMACOLOGICA SINICA
·
2021
·Vol. 43
·期 1
·121-132
·
IF 6.15
Zhao Ze-an,Jiang Yu,Chen Yan-yu,Wu Ting,Lan Qun-sheng,Li Yong-mei,Li Lu,Yang Yang,Lin Cui-ting,Cao Ying,Zhou Ping-zheng,Guo Jia-yin,Tian Yuan-xin,Pang Jian-xin
Urate transporter 1 (URAT1) and glucose transporter 9 (GLUT9) are important targets for the development of uric acid-lowering drugs. We previously showed that the flexible linkers of URAT1 inhibitors could enhance their potency. In this study we designed and synthesized CDER167, a novel RDEA3710 analogue, by introducing a linker (methylene) between the naphthalene and pyridine rings to increase flexibility, and characterized its pharmacological and pharmacokinetics properties in vitro and in vivo. We showed that CDER167 exerted dual-target inhibitory effects on both URAT1 and GLUT9: CDER167 concentration-dependently inhibited the uptake of [ 14 C]-uric acid in URAT1-expressing HEK293 cells with an IC 50 value of 2.08 ± 0.31 μM, which was similar to that of RDEA3170 (its IC 50 value was 1.47 ± 0.23 μM). Using site-directed mutagenesis, we demonstrated that CDER167 might interact with URAT1 at S35 and F365. In GLUT9-expressing HEK293T cells, CDER167 concentration-dependently inhibited GLUT9 with an IC 50 value of 91.55 ± 15.28 μM, whereas RDEA3170 at 100 μM had no effect on GLUT9. In potassium oxonate-induced hyperuricemic mice, oral administration of CDER167 (10 mg·kg −1 · d −1 ) for 7 days was more effective in lowering uric acid in blood and significantly promoted uric acid excretion in urine as compared with RDEA3170 (20 mg·kg −1 · d −1 ) administered. The animal experiment proved the safety of CDER167. In addition, CDER167 displayed better bioavailability than RDEA3170, better metabolic stability and no hERG toxicity at 100 μM. These results suggest that CDER167 deserves further investigation as a candidate antihyperuricemic drug targeting URAT1 and GLUT9.
研究领域:
药理学
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医学
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药物开发
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产品关联
DOI
PubMed
高影响因子
Frontiers in Materials
·
2022
·
IF 3.985
Chen Yu,Liu Miao
Application of amino-functionalized mesoporous silica-coated core-shell magnetic nanoparticles (Fe3O4@mSiO2-NH2 NPs) for adsorbing heavy metal ions has attracted intensive interest in recent years. Despite the cytotoxicity triggered by the co-exposure of nanoparticles (NPs) and metal ions in relatively high dosages being reported, the effect of the adsorbed heavy metal ions on the cytotoxicity to human cells remains unexplored. Herein, we demonstrated the effect of amino-functionalized Fe3O4@mSiO2 core-shell magnetic nanoparticles before and after adsorbing Pb2+ ions on the cytotoxicity of human kidney cells (HEK293). The surface morphology, viability, and oxidative stress (OS) induction of HEK293 cells incubated with Fe3O4@mSiO2-NH2 NPs and Pb2+ ion-adsorbed Fe3O4@mSiO2-NH2 NPs were assessed, respectively. Transmission electron microscopic (TEM) images of cell sections depicted that Fe3O4@mSiO2-NH2 NPs were internalized by HEK293 cells and gathered mainly in the cytoplasm. Cell viability (MTT) assays revealed the Fe3O4@mSiO2-NH2 NPs could enhance the cell viability to 119.9% and 108.2% compared to the control group, respectively. On contrast, the Pb2+ ion-adsorbed Fe3O4@mSiO2-NH2 NPs were toxic to the cell because when the Pb2+ ion contents were 5.0 and 7.5 μg mL−1, the viabilities of the samples decreased to 97.1% and 84.7%, respectively. Oxidative stress data proved that OS was negatively affected by both dissociative Pb2+ ions and the Pb2+ ion-adsorbed Fe3O4@mSiO2-NH2 NPs. Cytotoxicity may be attributed to the OS induced by Pb2+ ions leaked from the adsorbent. Under the same Pb2+ ion concentration, the cytotoxicity of the adsorbed Pb2+ ions was lower than that of the dissociative Pb2+ ions, indicating that the adsorption by NPs inhibited the cytotoxicity of Pb2+ ions. This work will provide new references for assessing the cytotoxicity of Pb2+-adsorbed nanoparticles.
研究领域:
纳米技术
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毒理学
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细胞生物学
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环境科学
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产品关联
DOI
高影响因子
INVESTIGATIONAL NEW DRUGS
·
2022
·Vol. 40
·期 5
·977-989
·
IF 3.651
Duan Zelin,Zhou Zhiyun,Lu Feifei,Zhang Yawen,Guo Xvqin,Gui Chunshan,Zhang Hongjian
Targeting tumor metabolic vulnerabilities such as “glutamine addiction” has become an attractive approach for the discovery of novel antitumor agents. Among various mechanisms explored, SLC1A5, a membrane transporter that plays an important role in glutamine cellular uptake, represents a viable target to interfere with tumor’s ability to acquire critical nutrients during proliferation. In the present study, a stably transfected HEK293 cell line with human SLC1A5 (HEK293-SLC1A5) was established for the screening and identification of small molecule SLC1A5 inhibitors. This in vitro system, in conjunction with direct measurement of SLC1A5-mediated L-glutamine-2,3,3,4,4-D 5 (substrate) uptake, was practical and efficient in ensuring the specificity of SLC1A5 inhibition. Among a group of diverse compounds tested, mianserin (a tetracyclic antidepressant) demonstrated a marked inhibition of SLC1A5-mediated glutamine uptake. Subsequent investigations using SW480 cells demonstrated that mianserin was capable of inhibiting SW480 tumor growth both in vitro and in vivo, and the in vivo antitumor efficacy was correlated to the reduction of glutamine concentrations in tumor tissues. Computational analysis revealed that hydrophobic interactions between SLC1A5 and its inhibitors could be a critical factor in drug design. Taken together, the current findings confirmed the feasibility of targeting SLC1A5-mediated glutamine uptake as a novel approach for antitumor intervention. It is anticipated that structural insights obtained based on homology modeling would lead to the discovery of more potent and specific SLC1A5 inhibitors for clinical development.
研究领域:
肿瘤学
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药理学
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分子生物学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
Journal of Neuroinflammation
·
2023
·Vol. 20
·期 1
·1-18
·
IF 9.3
Yao Ke,Mou Qianxue,Lou Xiaotong,Ye Meng,Zhao Bowen,Hu Yuanyuan,Luo Jing,Zhang Hong,Li Xing,Zhao Yin
Background Optic nerve injury (ONI) is a key cause of irreversible blindness and triggers retinal ganglion cells (RGCs) change and synapse loss. Microglia is the resistant immune cell in brain and retina and has been demonstrated to be highly related with neuron and synapse injury. However, the function of Sirtuin 1 (SIRT1), a neuroprotective molecule, in mediating microglial activation, retinal synapse loss and subsequent retinal ganglion cells death in optic nerve injury model as well as the regulatory mechanism remain unclear.MethodTo this end, optic nerve crush (ONC) model was conducted to mimic optic nerve injury. Resveratrol and EX527, highly specific activator and inhibitor of SIRT1, respectively, were used to explore the function of SIRT1 in vivo and vitro. Cx3Cr1-CreERT2/RaptorF/F mice were used to delete Raptor for inhibiting mammalian target of rapamycin complex 1 (mTORC1) activity in microglia. HEK293 and BV2 cells were transfected with plasmids to explore the regulatory mechanism of SIRT1.Results We discovered that microglial activation and synapse loss in retinal inner plexiform layer (IPL) occurred after optic nerve crush, with later-development retinal ganglion cells death. SIRT1 activation induced by resveratrol inhibited microglial activation and attenuated synapse loss and retinal ganglion cells injury. After injury, microglial phagocytosed synapse and SIRT1 inhibited this process to protect synapse and retinal ganglion cells. Moreover, SIRT1 exhibited neuron protective effects via activating tuberous sclerosis complex 2 (TSC2) through deacetylation, and enhancing the inhibition effect of tuberous sclerosis complex 2 on mammalian target of rapamycin complex 1 activity.Conclusion Our research provides novel insights into microglial SIRT1 in optic nerve injury and suggests a potential strategy for neuroprotective treatment of optic nerve injury disease.
研究领域:
神经科学
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眼科学
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分子生物学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
CHEMOSPHERE
·
2015
·
IF 3.34
Lu Lai,Jian-Cheng Jin,Zi-Qiang Xu,Ping Mei,Feng-Lei Jiang,Yi Liu
The toxicity of CdTe QDs with nearly identical maximum emission wavelength but modified with four different ligands (MPA, NAC, GSH and dBSA) to HEK293 and HeLa cells were investigated using flow cytometry, spectroscopic and microscopic methods. The results showed that the cytotoxicity of QDs increased in a dose- and time-dependent manner. No appreciable fraction of cells with sub-G1 DNA content, the loss of membrane integrity, and the swelling of nuclei clearly indicated that CdTe QDs could lead to necrotic cell death in HEK293 cells. JC-1 staining and TEM images confirmed that QDs induced MPT, which resulted in mitochondrial swelling, collapse of the membrane potential. MPT is an important step in QDs-induced necrosis. Moreover, QDs induced MPT through the elevation of ROS. The fluorimetric assay and theoretical analysis demonstrated ROS production has been associated with the internalization of QDs with cells. Due to large surface/volume ratios of QDs, when QDs added in the culture medium, serum proteins in the culture medium will be adsorbed on the surface of QDs. This adsorption of serum protein will change the surface properties and size, and then mediate the cellular uptake of QDs via the clathrin-mediated endocytic pathway. After entering into cells, the translocation of QDs in cells is usually via endosomal or lysosomal vesicles. The rapid degradation of QDs in lysosome and the lysosomal destabilization induce cell necrosis. This study provides a basis for understanding the cytotoxicity mechanism of CdTe QDs, and valuable information for safe use of QDs in the future.
研究领域:
毒理学
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细胞生物学
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材料科学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2018
·Vol. 8
·期 58
·33000-33009
·
IF 2.936
Xiaohong Chu,Ming Zhang,Ninglin Zhou,Fan Wu,Baohong Sun,Jian Shen
A novel antibacterial agent was synthesized using 2-(dimethylamino)ethyl methacrylate (DM) and sodium 3-chloro-2-hydroxypropane sulfonate (CHPS). It was characterized by Fourier transform infrared spectroscopy (FTIR), NMR Spectroscopy (1H NMR), and X-ray photoelectron spectroscopy (XPS). This new agent DMCHPS was then grafted onto a polyurethane (PU) substrate via surface-initiated reverse atom transfer radical polymerization (SI-RATRP). The modified PU was characterized by FTIR and XPS. The hydrophilic properties of the PU surface before and after the incorporation of DMCHPS were determined by static contact angle (SCA) measurements. The results showed that the hydrophilicity of the PU surface after the modification was remarkably improved. MIC tests and bacterial adhesion confirmed that modified PU has good antibacterial properties. Protein adsorption experiments show that the material has a certain ability to resist pollution. Furthermore, the high survival rate of HEK293 human embryonic kidney cells shows that the modified PU has a potential use as a medicinal material.
研究领域:
材料科学
·
高分子化学
·
生物医学工程
·
生物材料
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）