NEUROCHEMICAL RESEARCH
·
2021
·Vol. 47
·期 3
·713-722
·
IF 3.996
Zhang Haiju,Yu Shiqian,Xia Liping,Peng Xia,Wang Shun,Yao Baozhen
Epilepsy (SE) is a common and serious neurological disease. NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome participates in the pathogenesis of SE, while its underlying mechanism is still unclear. Here, we attempted to explore the mechanism of action of NLRP3 inflammasome in SE. SE mouse model was constructed by administration of kainic acid (KA). Astrocytes were treated with KA to mimic SE cell model. MCC950 (NLRP3 inhibitor) and Z-YVAD-FMK (Caspase-1 inhibitor) were used to treat astrocytes to inhibit the activity of NLRP3 and Caspase-1. Nissl staining was performed to examine the morphology of neuron. Western blot, enzyme-linked immunosorbent assay and immunofluorescence staining were performed to assess protein expression. SE mouse model exhibited an increase of neuronal loss, and an up-regulation of Cleaved-Caspase-1, IL-1β and IL-18 in hippocampus. The levels of GFAP + ADK + cells were significantly increased in SE mice. MCC950 or Z-YVAD-FMK abolished these impacts conferred by KA in SE mice. Moreover, KA treatment enhanced the expression of NLRP3, Cleaved-Caspase-1, IL-1β and IL-18 in astrocytes, which was rescued by knockdown of NLRP3 or Caspase-1. Additionally, CREB, p-CREB, REST were up-regulated, and SP1 was down-regulated in the KA-treated SE mice and KA-treated astrocytes. Inhibition of NLRP3 or Caspase-1 rescued these proteins expression in KA-treated astrocytes. CREB or REST silencing reduced adenosine kinase (ADK) expression, while SP1 knockdown enhanced ADK expression in KA-treated astrocytes. In conclusion, NLRP3 inflammasome activation enhances ADK expression to accelerate SE in mice through regulating CREB/REST/SP1 signaling pathway. Thus, inhibition of NLRP3 inflammasome may be a treatment for SE.
研究领域:
神经科学
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免疫学
·
药理学
·
细胞生物学
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产品关联
DOI
PubMed
高影响因子
BIOMACROMOLECULES
·
2025
·
IF 5.4
Yuduo Gao,Luwei Zhang,Ziyue Wang,Hao Bai,Chengfan Wu,Qi Shuai,Yunfeng Yan
mRNA-based therapies hold tremendous promise for treating various diseases, yet their clinical success is hindered by delivery challenges. This study developed a library of 140 lipocationic Poly(β-amino ester)s (PBAEs) and formulated lipid-polymer hybrid nanoparticles (LPHs) with four helper lipids, including 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), to enhance mRNA delivery. Initial in vitro screening of four representative PBAEs identified the D/P4-1 formulation (DOTAP/PBAE molar ratio of 4:1) as the most effective. Further screening of the library using this formulation identified eight top-performing LPHs. In vivo experiments confirmed high luciferase expression in the spleen and lungs of mice following intravenous administration of Luc mRNA-loaded LPHs. Detailed analysis revealed that DOTAP incorporation influenced LPH properties, including apparent pKa, surface charge, and internal hydrophobicity, enabling enhanced mRNA release and cellular uptake. This study demonstrates potent approaches to modulate PBAE-lipid nanoparticle properties by altering PBAE structures and nanoparticle composition, offering insights for designing effective hybrid carriers for mRNA therapeutics.
研究领域:
药学
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纳米技术
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分子生物学
·
治疗学
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产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2021
·
IF 3.978
Yiwei Dai,Chenchen Li,Luhua Zheng,Bo Jiang,Tao Zhang,Jingjing Chen
In our previous study, a whole-cell biocatalytic system involving α-glucan phosphorylase , phosphoglucomutase , glucose 6-phosphate isomerase, tagatose 1,6-bisphosphate aldolase , and phosphoglycolate phosphatase , was constructed to produce d -tagatose from maltodextrin . However, the biotransformation efficiency of this whole-cell catalyst was low because of the unbalanced ratio of each enzyme and metabolic flux of the intermediates. Therefore, in this study, the biosynthesis of d -tagatose was enhanced by optimizing vectors and improving the expression of rate-limiting enzymes through the construction of multi-copy genes to regulate expression levels. The conversion rate increased from 20.8% after 24 h to 25.2% after 3 h using 10 g/L maltodextrin as the substrate. Furthermore, the genes in the bypass pathways in Escherichia coli , pfka and zwf , were deleted to increase the accumulation of intermediates. The strain ER-2GatZ (Δ p Δ z ) produced 3.383 g/L d -tagatose using 10 g/L substrate after 3 h, which was 1.34 times that of the wild strain. This work exemplifies the use of modular engineering to enhance d -tagatose production using whole-cell catalysts.
研究领域:
生物技术
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代谢工程
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微生物学
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生化工程
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DOI
高影响因子
RSC Advances
·
2016
·Vol. 6
·期 54
·49250-49262
·
IF 3.289
Jilai Tian,Miaomiao Han,Yue Wang,Kang Qian,Xue Ke,Tianyuan Ci
For oncotherapy, a delivery system that can effectively prevent and attenuate tumor metastasis is greatly desirable. Herein, we report a novel delivery system of liposomal doxorubicin with a reduction-responsive modification of low molecular weight heparin (LMWH) to realize higher efficacy in the prevention of tumor metastasis. A series of DOX liposomal formulations of crude DOX-Lip, reduction-insensitive LMWH-DOX-Lip and reduction-responsive LMWH-ss-DOX-Lip were prepared. The reduction-responsive modification of DOX liposomes with LMWH chains resulted in higher cellular uptake, higher cytotoxicity of DOX and higher anti-invasion and anti-migration abilities of LMWH compared with reduction-insensitive liposomes, owing to the fast release of DOX and LMWH in the tumor cytoplasm. Meanwhile, the evaluation of heparanase expression was also adopted to reveal the possible underlying mechanism of the effects of LMWH on the attenuation of tumor metastasis. In vivo pulmonary melanoma metastasis assays confirmed the enhanced drug efficacy and lower side effects of LMWH-ss-DOX-Lip compared with crude DOX-Lip or the reduction-insensitive control group. These results may be attributed to the rational design of the drug carriers with a synergistic combination of LMWH moieties and DOX loaded liposomes via reduction-responsive modification.
研究领域:
药学
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肿瘤学
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药物递送系统
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癌症治疗
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产品关联
DOI
高影响因子
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
·
2017
·Vol. 33
·期 2
·1-9
·
IF 1.658
Zhang Jian-Fen,Chen Wei-Qing,Chen Hong
In this study, we report the cloning and expression of a functional glucoside 3-dehydrogenase (G3DH) gene from Sphingobacterium faecium ZJF-D6. This gene is 1686 bp in length and encodes a peptide of 562 amino acids. The G3DH gene was successfully expressed in E. coli , and the recombinant enzyme could oxidize glucosides, galactosides and analogues at C-3 position. The sequence and multiple alignment analysis showed that the enzyme has highest identity with G3DHs from Paraglaciecola polaris LMG 21857, Aliiglaciecola lipolytica E3 and Halomonas sp. alpha-15. The recombinant G3DH was purified on Ni–NTA column and exhibited the highest activity at pH 7.6 and 30 °C. It was sensitive to acid and alkali, and showed well thermostability. The SfG3DH could oxidize a wild range of sugars. When recombinant E. coli BL21 cells were used as catalyst, a high rate of conversion to N- p -nitrophenyl-3-ketovalidamine was achieved, and no p -nitroaniline was detected. This process offers a promising approach to fulfill substrate of 3-ketovalidoxylamine A C–N lyase production.
研究领域:
微生物学
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生物化学
·
酶学
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生物催化
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DOI
PubMed
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2022
·Vol. 45
·期 8
·1319-1329
·
IF 3.434
Yi Xia,Wu Jianfang,Jiang He,Zhao Yan,Mei Jun
Kinases modulate the various physiological activities of microbial fermenting strains including the conversion of lignocellulose-derived phenolic aldehydes (4-hydroxyaldehyde, vanillin, and syringaldehyde). Here, we comprehensively investigated the gene transcriptional profiling of the kinases under the stress of phenolic aldehydes for ethanologenic Zymomonas mobilis using DNA microarray. Among 47 kinase genes, three genes of ZMO0003 (adenylylsulfate kinase), ZMO1162 (histidine kinase), and ZMO1391 (diacylglycerol kinase), were differentially expressed against 4-hydroxybenzaldehyde and vanillin, in which the overexpression of ZMO1162 promoted the phenolic aldehydes conversion and ethanol fermentability. The perturbance originated from plasmid-based expression of ZMO1162 gene contributed to a unique expression profiling of genome-encoding genes under all three phenolic aldehydes stress. Differentially expressed ribosome genes were predicted as one of the main contributors to phenolic aldehydes conversion and thus finally enhanced ethanol fermentability for Z. mobilis ZM4. The results provided an insight into the kinases on regulation of phenolic aldehydes conversion and ethanol fermentability for Z. mobilis ZM4, as well as the target object for rational design of robust biorefinery strains.
研究领域:
微生物学
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生物技术
·
分子生物学
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生物化学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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