MEDICINAL CHEMISTRY RESEARCH
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2020
·Vol. 29
·期 7
·1230-1237
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IF 1.783
Jiang Tingting,Cheng Ting,Li Jing,Zhou Mengyue,Tan Rong,Yang Xiaojing,Wang Yang,Li Weiwei,Zheng Jiang
Grapefruit-mediated fruit–drug interaction has been well discussed in clinical practice. However, the biochemical mechanisms for such interaction are far beyond our understanding. The objectives of the present study aimed at the interaction between bergaptol (BGT) and CYP2C9. BGT, one of the major furanocoumarin constituents in grapefruit, has been reported to inhibit the activity of CYP2C9. And a number of furan-containing compounds are known to act as mechanism-based inactivators of CYP2C9. In this study, BGT was found to induce time-, concentration-, and NADPH-dependent irreversible inhibition of CYP2C9. A GSH conjugate was detected in an incubation mixture containing CYP2C9, BGT, NADPH, and GSH. Further mechanistic investigation revealed that a γ-ketoenal metabolite was responsible for the enzyme inactivation. The obtained data indicate that BGT was a mechanism-based inactivator of CYP2C9. The study would facilitate the understanding of the mechanistic insight into fruit–drug interactions mediated by grapefruits.
研究领域:
药理学
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生物化学
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临床药理学
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DOI
高影响因子
JOURNAL OF CELLULAR BIOCHEMISTRY
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2019
·Vol. 120
·期 10
·18509-18523
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IF 3.448
Yujie Yao,Xueyuan Hu,Xiujing Feng,Yuan Zhao,Manyu Song,Chaoran Wang,Honggang Fan
Dexmedetomidine (DEX) prevents kidney damage caused by sepsis, but the mechanism of this effect remains unclear. In this study, the protective molecular mechanism of DEX in lipopolysaccharide (LPS)-induced acute kidney injury was investigated and its potential pharmacological targets from the perspective of inhibiting oxidative stress damage and the nucleotide-binding domain-like receptor protein 3 (NLRP3) inflammasome activation. Intraperitoneal injection of DEX (30 μg/kg) significantly improved LPS (10 mg/kg) induced renal pathological damage and renal dysfunction. DEX also ameliorated oxidative stress damage by reducing the contents of reactive oxygen species, malondialdehyde and hydrogen peroxide, and increasing the level of glutathione, as well as the activity of superoxide dismutase and catalase. In addition, DEX prevented nuclear factor-kappa B (NF-κB) activation and I-kappa B (IκB) phosphorylation, as well as the expressions of NLRP3 inflammasome-associated protein and downstream IL-18 and IL-1β. The messengerRNA (mRNA) and protein expressions of toll-like receptor 4 (TLR4), NADPH oxidase-4 (NOX4), NF-κB, and NLRP3 were also significantly reduced by DEX. Their expressions were further evaluated by immunohistochemistry, yielding results were consistent with the results of mRNA and protein detection. Interestingly, the protective effects of DEX were reversed by atipamezole-an alpha 2 adrenal receptor (α 2 AR) inhibitor, whereas idazoxan-an imidazoline receptor (IR) inhibitor failed to reverse this change. In conclusion, DEX attenuated LPS-induced AKI by inhibiting oxidative stress damage and NLRP3 inflammasome activation via regulating the TLR4/NOX4/NF-κB pathway, mainly acting on the α 2 AR rather than IR.
研究领域:
药理学
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免疫学
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肾脏生理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Nature Protocols
·
2018
·Vol. 13
·期 10
·2362-2386
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IF 12.423
Zou Yejun,Wang Aoxue,Shi Mei,Chen Xianjun,Liu Renmei,Li Ting,Zhang Chenxia,Zhang Zhuo,Zhu Linyong,Ju Zhenyu,Loscalzo Joseph,Yang Yi,Zhao Yuzheng
Cellular oxidation–reduction reactions are mainly regulated by pyridine nucleotides (NADPH/NADP + and NADH/NAD + ), thiols, and reactive oxygen species (ROS) and play central roles in cell metabolism, cellular signaling, and cell-fate decisions. A comprehensive evaluation or multiplex analysis of redox landscapes and dynamics in intact living cells is important for interrogating cell functions in both healthy and disease states; however, until recently, this goal has been limited by the lack of a complete set of redox sensors. We recently reported the development of a series of highly responsive, genetically encoded fluorescent sensors for NADPH that substantially strengthen the existing toolset of genetically encoded sensors for thiols, H 2 O 2 , and NADH redox states. By combining sensors with unique spectral properties and specific subcellular targeting domains, our approach allows simultaneous imaging of up to four different sensors. In this protocol, we first describe strategies for multiplex fluorescence imaging of these sensors in single cells; then we demonstrate how to apply these sensors to study changes in redox landscapes during the cell cycle, after macrophage activation, and in living zebrafish. This approach can be adapted to different genetically encoded fluorescent sensors and various analytical platforms such as fluorescence microscopy, high-content imaging systems, flow cytometry, and microplate readers. A typical preparation of cells or zebrafish expressing different sensors takes 2–3 d; microscopy imaging or flow-cytometry analysis can be performed within 5–60 min.
研究领域:
细胞生物学
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分子生物学
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生物化学
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DOI
PubMed
高影响因子
Liver Research
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2021
·
IF 0
Guizhi Yang,Shengwen Li,Jiahua Jin,Yuanyuan Xuan,Liqin Ding,Minxia Huang,Jun Liu,Biye Wang,Tian Lan
Background and aim Liver fibrosis resulting from persistent liver injury represents a major healthcare problem globally. Traditional Chinese medicine has played an essential role in the treatment of liver fibrosis in recent years. Thus, this study aims to assess the effect of Longhu Rendan (LHRD), a Chinese traditional patent medicine, on liver fibrosis and its potential mechanism. Methods The liver fibrosis in mice was induced via the intraperitoneal injection of carbon tetrachloride (CCl 4 ) for 6 weeks or bile duct ligation for 15 days. Various methods were used to judge the therapeutic effect of LHRD. Results LHRD significantly suppressed the activity of serum index of abnormal liver function, liver cell apoptosis, and necrosis, attenuating liver injury. Moreover, LHRD treatment alleviated liver fibrotic features, such as the reduction of collagen deposition and hepatic stellate cell activation as well as profibrotic gene expression. Mechanistically, LHRD treatment inhibited nuclear transcription factor-kappa B signaling and inflammatory gene expression and diminished the production of reactive oxygen species and 4-hydroxynonenal, along with the downregulation of NADPH oxidase 4. Conclusions Overall, the present study demonstrates that LHRD ameliorates liver injury and fibrosis via the inhibition of inflammation and oxidative stress in mice, indicating that LHRD is a potential medicine for the treatment of liver fibrosis.
研究领域:
药理学
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肝病学
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中医药
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DOI
Journal of Pharmaceutical Analysis
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2024
·
IF 6.1
Caiyi Yang,Kehan Chen,Yunliang Chen,Xuting Xie,Pengcheng Li,Meng Zhao,Junjie Liang,Xueqian Xie,Xiaoyun Chen,Yanping Cai,Bo Xu,Qing Wang,Lian Zhou,Xia Luo
The incomplete degradation of tumour cells by macrophages (Mφ) is a contributing factor to tumour progression and metastasis, and the degradation function of Mφ is mediated through phagosomes and lysosomes. In our preliminary experiments, we found that overactivation of NADPH oxidase 2 (NOX2) reduced the ability of Mφ to degrade engulfed tumour cells. Above this, we screened out liquiritin from Glycyrrhiza uralensis Fisch , which can significantly inhibit NOX2 activity and inhibit tumours, to elucidate that suppressing NOX2 can enhance the ability of Mφ to degrade tumour cells. We found that the tumour environment could activate the NOX2 activity in Mφ phagosomes, causing Mφ to produce excessive reactive oxygen species (ROS), thus prohibiting the formation of phagolysosomes before degradation. Conversely, inhibiting NOX2 in Mφ by liquiritin can reduce ROS and promote phagosome-lysosome fusion, therefore improving the enzymatic degradation of tumour cells after phagocytosis, and subsequently promote T cell activity by presenting antigens. We further confirmed that liquiritin down-regulated the expression of the NOX2 specific membrane component protein gp91 phox, blocking its binding to the NOX2 cytoplasmic component proteins p67 phox and p47 phox, thereby inhibiting the activity of NOX2. This study elucidates the specific mechanism by which Mφ cannot degrade tumour cells after phagocytosis, and indicates that liquiritin can promote the ability of Mφ to degrade tumour cells by suppressing NOX2.
研究领域:
免疫学
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癌症生物学
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细胞生物学
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药理学
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DOI
PubMed
高影响因子
Nature Communications
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2025
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IF 15.7
Jiang Zhentao,Rao Jingxin,Zhu Caokai,Li Yamiao,Zhu Qiang,Zheng Mingyue,Zhang Wenchi,Zhang Rongzhen
Bilirubin biosynthesis has long been constrained by low yields and poorly understood bottlenecks. Here, we report a fully in vitro pathway that converts heme to bilirubin with the titer of 1.7 g/L and 95.8%. Systematically, enzyme screening and mechanistic analysis reveal the hidden challenge: Fe²⁺-induced oxidative degradation of intermediates. We show that Fe²⁺ coordinates with deprotonated intermediates to trigger oxidative ring-opening degradation via O₂-mediated radical mechanism, as supported by DFT calculations indicating reduced HOMO-LUMO gap in Fe²⁺-ligand complexes. The degradation is mitigated through competitive iron chelation and protonation state modulation, improving yield to 80.1%. Furthermore, we have resolved heme-CO complexes blocking O₂ activation at heme oxygenase by introducing a carbon monoxide dehydrogenase to remove CO and restore enzyme activity. Coupled with NADPH-recycling via formate dehydrogenase, these interventions enable efficient, scalable bilirubin synthesis with a 20-fold improvement. Our work shows controlling inhibitory byproducts is critical for stabilizing heme-related pathways and as a generalizable framework for synthetic biology.
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DOI
PubMed
高影响因子
METABOLIC ENGINEERING
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2025
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IF 6.8
Xin Jin,Ruxin Hao,Hannuo Shen,Zhu Liu,Sumeng Wang,Qingsheng Qi,Quanfeng Liang
The design of synthetic driving forces for biosynthetic pathway is crucial for directing carbon flux toward the target product. Optimizing cellular redox status is one of the key strategies for constructing microbial cell factories. In this study, we attempt to create a novel redox imbalance force-driven (RIFD) strategy to direct carbon flow toward the target synthetic pathway. Initially, we increased the NADPH pool through a strategy of “open source and reduce expenditure” employing four approaches to achieve excessive NADPH levels and growth inhibition: (I) the expression of cofactor-converting enzymes, (II) the expression of heterologous cofactor-dependent enzymes, (III) the expression of enzymes involved in the NADPH synthesis pathway, and (IV) reduced NADPH wastage by knocking down non-essential genes that consume NADPH in vivo. Next, multiple automated genome engineering (MAGE) techniques were employed to evolve redox-imbalanced engineered strains and drive the metabolic flux to L-threonine production. Finally, we developed a NADPH and L-threonine dual-sensing biosensor, combined it with Fluorescence-Activated Cell Sorting (FACS), and a high-yield (0.65 g/g) L-threonine-producing strain with a titer of 117.65 g L −1 was obtained. This research presents a general approach to increasing the production of cofactor-related products. Utilizing redox imbalance forces to drive metabolic flow toward the target product, it is possible to increase production while simultaneously restoring cell growth.
研究领域:
合成生物学
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代谢工程
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系统生物学
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生物技术
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生物工程
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产品关联
DOI
PubMed
高影响因子
Heliyon
·
2024
·
IF 3.4
Qi Wu,Xue-li Ge,Zi-kai Geng,Hao Wu,Jing-yi Yang,Shi-rong Cao,Ai-lin Yang
HuaChanSu is active water extracts from the skin of Bufo bufo gargarizans Cantor. It has been already used to treat clinical cancers including HCC (Hepatocellular carcinoma, HCC), however, the molecular mechanisms under HuaChanSu's anti-cancer effects remain unclear. PPP (Pentose phosphate pathway, PPP), the major source of ribose and NADPH (Nicotinamide adenine dinucleotide phosphate, NADPH), is always over-activated and particularly critical for tumor cells growth. In this study, firstly, we illustrate that HuaChanSu restrains the growth of human hepatoma cells. More importantly, we demonstrate that the expression of G6PD (Glucose-6-phosphate dehydrogenase, G6PD), the first rate-limiting enzyme of the PPP, is restrained in human hepatoma cells after treatment with HuaChanSu. Additionally, our results show that G6PD enzyme activity and dimer formation are inhibited by HuaChanSu. Furthermore, we find that HuaChanSu could inhibit NADPH production and nucleotide level. In addition, we identify that expression of PLK1 (Polo-like kinase 1, PLK1) is also reduced in response to HuaChanSu, and knockdown of PLK1 restrains enzyme activity and dimer formation of G6PD, but has no effect on G6PD protein level. Subsequently, we demonstrate that inhibition of G6PD could restrain the proliferation of tumor cells and enhance the inhibitory effect of HuaChanSu on cell proliferation of human hepatoma cells. In conclusion, for the first time, our study reveals that HuaChanSu interferes with PPP via suppression of G6PD expression and enzyme activity to restrain growth of tumor cells, and these results provide a novel insight for the anti-hepatoma mechanisms of HuaChanSu and promote the innovation of the research model of TCM. Moreover, the development of drugs targeting abnormal tumor metabolism is currently a hot topic, our works provide theoretical support for further drug development from HuaChanSu, meanwhile, the revelation of the new molecular mechanism also provides a new perspective for the study of the pathogenesis of liver cancer.
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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癌症代谢
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2022
·
IF 5.895
Xiang Wang,Jin-Hua Mou,Zi-Hao Qin,Ting-Bin Hao,Lan Zheng,Joseph Buhagiar,Yu-Hong Liu,Srinivasan Balamurugan,Yuhe He,Carol Sze Ki Lin,Wei-Dong Yang,Hong-Ye Li
The unicellular freshwater green alga Haematococcus pluvialis has attracted much research attention due to its biosynthetic ability for large amounts of astaxanthin, a blood-red ketocarotenoid that is used in cosmetics, nutraceuticals, and pharmaceuticals. Recently, numerous studies have investigated the functions of natural astaxanthin; however, the high cost of the production of astaxanthin from H. pluvialis cultures restricts its commercial viability. There is an urgent need to fulfill commercial demands by increasing astaxanthin accumulation from H. pluvialis cultures. In this study, we discovered that treatment of H. pluvialis cultures at the beginning of the macrozooid stage (day 0) with 1 μM rac-GR24, a synthetic analogue of strigolactones (a class of phytohormones), led to significant increases in biomass [up to a maximum dry cell weight (DCW) of 0.53 g/L] during the macrozooid stage and astaxanthin (from 0.63 to 5.32% of DCW) during the hematocyst stage. We elucidated that this enhancement of biomass accumulation during the macrozooid stage by rac-GR24 is due to its increasing CO2 utilization efficiency in photosynthesis and carbohydrate biosynthesis. We also found that rac-GR24 stimulated the overproduction of nicotinamide adenine dinucleotide phosphate (NADPH) and antioxidant enzymes in H. pluvialis cultures, which alleviated the oxidative damage caused by reactive oxygen species generated during the hematocyst stage due to the exhaustion of nitrogen supplies. Moreover, rac-GR24 treatment of H. pluvialis synergistically altered the activity of the pathways of fatty acid biosynthesis and astaxanthin esterification, which resulted in larger amounts of astaxanthin being generated by rac-GR24-treated cultures than by controls. In summary, we have developed a feasible and economic rac-GR24-assisted strategy that increases the amounts of biomass and astaxanthin generated by H. pluvialis cultures, and have provided novel insights into the mechanistic roles of rac-GR24 to achieve these effects.
研究领域:
植物学
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植物化学
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生物技术
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药理学
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DOI
PubMed
高影响因子
Green Synthesis and Catalysis
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2022
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IF 0
Yongxing Li,Pengqian Luan,Lele Dong,Jianqiao Liu,Luying Jiang,Jing Bai,Fufeng Liu,Yanjun Jiang
Asymmetric reduction of the conjugated C C bonds by the old yellow enzymes (OYEs) presents a promising field in the synthesis of chiral chemicals. Nevertheless, few natural OYEs have been applied in large-scale applications due to the requirement of costly NADPH and low operational stability. Herein, a stable and efficient fusion of YqjM from Bacillus subtilis and glucose dehydrogenase (GDH) from Bacillus megaterium was constructed to stereoselectively reduce the conjugated C C bonds in a self-sufficient continuous process. The effects of the enzyme order and different linkers on the fusions were investigated by structural analysis and all-atom molecular dynamics simulation. The best fusion YqjM_G_GDH gave 98% conversion of 100 mmol/L 2-methylcyclopentenone with an excellent ee value (>99%) in 3 h, while the mixture of individual enzymes only obtained 68% conversion after more than 8 h. The improved substrate conversion of YqjM_G_GDH fusion was probably attributed to the increased flexibility of each fused enzyme and the shortening of the diffusion distance of NADPH regenerated. A one-pot process was designed to purify and immobilize the fusion on the Ni 2+ -nitrilotriacetic acid functionalized magnetic mesoporous silica nanoflowers. The resulting immobilized biocatalyst not only catalyzed the asymmetric reduction of various α,β -unsaturated ketones (20 mmol/L) continuously with only 50 μmol/L NADP + to initiate the whole process, but also retained more than 82% of the initial activity after seven cycles, serving as a good candidate for the industrial applications.
研究领域:
生物催化
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酶工程
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有机化学
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材料科学
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DOI
BIOMACROMOLECULES
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2018
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IF 5.738
Xue-Jian Zhang,Xiao-Wei Wang,Jia-Xing Sun,Chao Su,Shuguang Yang,Wen-Bin Zhang
Protein immobilization is critical to utilize their unique functions in diverse applications. Herein, we report that orthogonal peptide–protein chemistry enabled multilayer construction can facilitate the incorporation of various folded structural domains, including calmodulin in different states, affibody, and dihydrofolate reductase (DHFR). An extended conformation is found to be the most advantageous for steady film growth. The resulting protein thin films exhibit sensitive and selective responsive behaviors to biosignals, such as Ca2+, trifluoperazine, and nicotinamide adenine dinucleotide phosphate (NADPH), and fully maintain the catalytic activity of DHFR. The approach is applicable to different substrates such as hydrophobic gold and hydrophilic silica microparticles. The DHFR enzyme can be immobilized onto silica microparticles with tunable amounts. The multilayer setup exhibits a synergistic enhancement of DHFR activity with increasing numbers of bilayers and also makes the embedded DHFR more resilient to lyophilization. Therefore, this is a convenient and versatile method for protein immobilization with potential benefits of synergistic enhancement in enzyme performance and resilience.
研究领域:
生物化学
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材料科学
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生物技术
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酶学
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DOI
PubMed
高影响因子
ACS Nano
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2020
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IF 14.588
Xuliang Guo,Fang Liu,Jian Deng,Peipei Dai,Yan Qin,Zhi Li,Bingbing Wang,Aiping Fan,Zheng Wang,Yanjun Zhao
Ferroptotic antitumor therapy has been compromised by various intracellular antioxidants, particularly glutathione and thioredoxin. Both are cofactors of glutathione peroxide 4 (GPX4) that act against oxidative stress via catalyzing the reduction of lipid peroxides. It was postulated that tailored polymer micelles could enhance ferroptotic antitumor efficacy via diminishing glutathione and thioredoxin under hypoxia. The aim was to engineer hypoxia-responsive micelles for selective enhancement of ferroptotic cell death in solid tumor. The polymer contains hydrophilic poly(ethylene glycol) (PEG) that is linked by azobenzene linker with nitroimidazole-conjugated polypeptide. The tailored polymer could self-assemble into nanoscale micelles to encapsulate RAS-selective lethal small molecule 3, a covalent GPX4 inhibitor. Under hypoxia, the azobenzene moiety enabled PEG shedding and enhanced micelles uptake in 4T1 cells. Likewise, the nitroimidazole moiety was reduced by the overexpressed nitroreductase with reduced nicotinamide adenine dinucleotide phosphate (NADPH) as the cofactor, resulting in transient depletion of NADPH. This impaired both the glutathione and thioredoxin redox cycle, leading to diminished intracellular glutathione and thioredoxin. The selective potency of ferroptotic micelles in depleting NADPH, glutathione and thioredoxin was further verified in vivo in the 4T1 tumor xenograft mice model. This work highlights the role of hypoxia-responsive polymers in enhancing the potency of ferroptotic inducers against solid tumors without additional side effects to healthy organs.
研究领域:
肿瘤学
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药学
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生物医学工程
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癌症生物学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）