Separation Science Plus
·
2023
·Vol. 6
·期 3
·2200120
·
IF 1.1
Zhe Hou,Lei Zhang,Xuan Guo,Hui Li,Xiaodong Li,Jing Dong,Guangqin Zhang,Dongyang Liu
The aim of this study was to develop a validated liquid chromatography–tandem mass spectrometry method to quantify nine important transporter proteins, so as to support the quantification of transporter proteins abundance in different human tissues and organs. Quantification was performed by targeted proteomics in positive multiple reaction monitoring mode with liquid chromatography–tandem mass spectrometry. The method was validated with respect to specificity, linearity, accuracy, precision, and stability. All the correlation coefficients (r) were larger than 0.99 and the validation ranges of multidrug-resistant transporter 1, breast cancer resistance protein, organic cation transporter 2, and organic anion transporter 1 were 0.036–3.636 nM, organic anion transporter 3 was 0.018–1.818 nM, organic anion transporting polypeptide transporter 1B1 and multidrug and toxin extrusion transporter 1 were 0.091–9.091 nM, organic anion transporting polypeptide transporter 1B3 and multidrug and toxin extrusion transporter 2K were 0.182–18.182 nM. The results of intra-day and inter-day accuracy and precision were all in line with the quantification requirements of biological samples and the stability results indicated that the samples were stable under current processing and storage conditions. The method is sensitive and efficient for the quantification of nine transporters in human tissues.
研究领域:
药理学
·
分析化学
·
蛋白质组学
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产品关联
DOI
高影响因子
Environmental Science-Processes & Impacts
·
2024
·Vol. 26
·期 7
·1171-1183
·
IF 4.3
Wenxuan Wang,Haiou Wang,Xiaoyu Ren,Wenxiao Zhang,Qian Li
To explore key factors involved in the uptake, translocation and accumulation of organophosphate esters (OPEs), computer simulation analysis and hydroponic experiments were executed. Lipid transporters with stocky-like active (SAC) cavities usually showed stronger binding affinities with the OPEs, especially when the SAC cavities belong to the Fish Trap model according to molecular docking. In our hydroponic trial, the binding affinity and gene expression of the lipid transporters and log Kow of the OPEs could be charged to the uptake, translocation and accumulation of the OPEs; however, these three factors played various important roles in roots and shoots. In detail, the effect of gene expression and binding affinity were stronger than log Kow in roots uptake and accumulation, but the result was the opposite in the shoots translocation. Transporters OsTIL and OsLTPL1 among all investigated transporters could play key roles in transporter-mediated OPE uptake, translocation and accumulation in the roots and shoots. OsMLP could be involved in the bidirected vertical translocation of the OPEs. OsLTP2 and OsLTP4 mainly acted as transporters of the OPEs in roots.
研究领域:
环境科学
·
植物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL SCIENCES
·
2019
·
IF 3.197
Yuqing Wang,Ping Li,Feifeng Song,Xi Yang,Yayun Weng,Zhiyuan Ma,Lu Wang,Huidi Jiang
PHT2, a member of the proton-coupled oligopeptide transporter family, participates in the transportation of small peptides and histidine from lysosomes to the cytosol. It facilitates maintenance of intracellular peptide homeostasis. However, it remains a challenge to elucidate the functional properties of PHT2 due to its localization in the lysosomal membrane. The aim of this study was to explore the transport function and substrate properties of human PHT2 (hPHT2) by transfecting Madin-Darby canine kidney cells with hPHT2 mutants to obtain stably expressed protein in the cell membrane. Using this cell model, we found that the transport activity of hPHT2 reached a maximum capacity when the extracellular pH was 5.5. hPHT2 showed relatively low affinity for Gly-Sar and relatively high affinity for d 3 -L-histidine, with K m values of 428 ± 88 μM and 66.9 ± 5.7 μM, respectively. Several typical substrates or inhibitors of PEPT1 and PEPT2, including valacyclovir , Gly-Gly-Gly, and cefadroxil but not 5-aminolevulinic acid or captopril , were proven to be substrates of hPHT2. However, hPHT2 showed low affinity for valacyclovir with a K m value of 5350 ± 1234 μM. In conclusion, this study established a suitable and efficient cell model to explore the function of hPHT2 in vitro and provided important information on the transport activity and substrate properties of hPHT2.
研究领域:
分子生物学
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药理学
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细胞生理学
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产品关联
DOI
PubMed
高影响因子
XENOBIOTICA
·
2021
·
IF 1.908
Jadera Talap,Zhuowei Shen,Jing Nie,Jie Pan,Mingcheng Xu,Kui Zeng,Kaifeng He,Fengting Ou,Houhong He,Jianbiao Yao,Ruwei Wang,Lushan Yu,Su Zeng
6-Hydroxykynurenic acid (6-HKA) is a nitrogen-containing phenolic acid compound in Ginkgo biloba leaves. The pharmacological activities of 6-HKA have been reported and shown that 6-HKA has the potential to become a therapeutic drug and may play an important role in the treatment of nervous system diseases. However, there are few studies on the drug metabolism and transport of 6-HKA. The aim of this study is to investigate the in vitro metabolism of 6-HKA and its interaction with multiple important drug transporters.The in vitro metabolism experiments in the present study demonstrate that 6-HKA might not undergo phase-I or phase-II metabolism in hepatic microsomes/S9 of rats. In addition, some drug transporters, including OAT1/3, OCT2, MDR1, OATP1B1, MATE1/2K and OCTN2, were investigated. The cellular uptake assays indicate that 6-HKA exhibits inhibition to the transport of classical substrates mediated by OAT3, OCT2, MATE2K and OCTN2 but has no significant effect on the transport of substrates mediated by MDR1, OAT1, OATP1B1 or MATE1. Further investigation of cellular accumulation assays shows that 6-HKA might be the substrate of OAT3, but not OCT2 or OCTN2. The bidirectional transport study suggests that 6-HKA is not a substrate of MDR1.The information about the in vitro metabolism of 6-HKA and the interaction between 6-HKA and some transporters will help us to better understand the pharmacokinetic properties of 6-HKA and provide reference for its pharmacodynamics, DDIs and drug-food interactions studies.
研究领域:
药理学
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药物代谢动力学
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生物化学
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产品关联
DOI
PubMed
高影响因子
Food & Function
·
2019
·Vol. 10
·期 8
·5215-5227
·
IF 3.241
Yufeng Chen,Fan Xue,Guobin Xia,Zhenlei Zhao,Chun Chen,Yunhong Li,Ying Zhang
7,8-Dihydroxyflavone (7,8-DHF), as a high-affinity TrkB receptor agonist, has been extensively explored in many human disorders involving brain-derived neurotrophic factor (BDNF) such as Alzheimer's disease, Parkinson's disease, depression, and obesity. However, to date, the transepithelial transport mechanisms of 7,8-DHF in the intestines remain unclear. The aim of our work was to quantify and to characterize in vitro transport of naturally occurring 7,8-DHF distinguished by its physicochemical and pharmacological properties. We discussed the transport mechanisms of 7,8-DHF using the Caco-2 cell model to determine the bi-directional permeability with different environmental factors (time, concentration, pH, metabolic inhibitors etc.). The influx and efflux characteristics of 7,8-DHF were also clarified. 7,8-DHF was poorly transported across Caco-2 cell monolayers by mainly passive diffusion via a transcellular pathway and not a paracellular pathway. The transport of 7,8-DHF was time and concentration-dependent in both the apical (AP) to basolateral (BL) side and the reverse direction. Interestingly, decreasing the pH from 7.4 to 6.0 markedly enhanced 7,8-DHF transport. It is noteworthy that 7,8-DHF transport was strongly inhibited by metabolic inhibitors and was highly dependent on temperature. The efflux ratio (ER) values at different concentrations were all above 1.5, indicating the existence of the efflux transporter. We found that breast cancer resistance protein (BCRP) was not involved in 7,8-DHF secretion and that the transport mechanism of 7,8-DHF was passive transport with an active efflux mediated by P-glycoprotein (P-gp) and multidrug resistance associated proteins (MRPs), particularly MRP 2. Moreover, the use of various influx transporter inhibitors in Caco-2 cells showed that organic cation transporters (OCTs) and organic anion-transporting polypeptides (OATPs) participated in 7,8-DHF transport. Taken together, the elucidated transport characteristics of 7,8-DHF provide useful information for designing novel and efficient delivery systems and avoiding food–food or food–drug interactions.
研究领域:
药理学
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细胞生物学
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神经科学
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药物递送系统
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF RADIOANALYTICAL AND NUCLEAR CHEMISTRY
·
2014
·Vol. 301
·期 1
·257-262
·
IF 1.415
Zha Zhongyong,Wang Dingna,Hong Wei,Liu Li,Zhou Sai,Feng Xiaojie,Qin Bing,Wang Jianmei,Yang Yi,Du Liang,Zhang Dong,Fang Zhendong,Xia Chuanqin
This study aimed at influence of europium speciation on its accumulation in Brassica napus (CK) and over-expressing BnTR1 lines (OE), and the kinetics of Eu uptake were investigated. These results indicated that the uptake in the roots of OE was higher than that of CK at high concentrations of europium and even the enrichment in the roots of OE was as high as 20,000 mg/kg at 328.9 μΜ Eu, this suggested that OE might have better resistance to uranium than CK. The time kinetics in plants showed that there had the similar trend between CK and OE. The formation of Eu 3+ at pH 5 in deionized water was beneficial to plants enrichment Eu and carbonate could reduce the adsorption of Eu in two group and phosphate almost completely impress the adsorption of Eu in two group, but citric acid could enhance europium root-to-shoot translocation in two group. These results would help understanding the mechanism of Eu uptake in Brassica napus (CK) and over-expressing BnTR1 lines (OE), therefore properly developing efficient europium/americium phytoremediation.
研究领域:
植物学
·
环境科学
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DOI
高影响因子
INTERNATIONAL JOURNAL OF PHARMACEUTICS
·
2020
·
IF 4.845
Shan Huang,Yike Huang,Xiongwei Yin,Dandan Wang,Wei Xiang,Min Wang,Zhining Xia
The absorption mechanisms of drugs play an important role on development and various application of drugs. However, the present methods of absorption mechanisms are not perfect enough. Hence, exploring a novel method to accurately predict the absorption mechanisms is necessary. In this study, we developed an Ussing model coupled with artificial membrane (AM-Ussing) for predicting the permeability of passive diffusion. Furthermore, by the combination of AM-Ussing and Ussing model, the three different mechanisms including efflux transport, uptake transport and passive diffusion could be successfully distinguished. Specifically, 12 drugs of passive transport were selected to evaluate AM-Ussing. And the high permeability correlation between AM-Ussing model and traditional Ussing model indicated that AM-Ussing was successfully established. Moreover, the absorption mechanisms of 11 drugs were predicted by the combination of AM-Ussing and Ussing model. The results showed that three kinds of absorption mechanisms mentioned can be predicted successfully. Therefore, AM-Ussing model is a potential tool to assess passive diffusion, and the combined use of AM-Ussing and Ussing model is an effective method to predict the intestinal absorption mechanisms of drugs.
研究领域:
药剂学
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药理学
·
生物药剂学
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产品关联
DOI
PubMed
高影响因子
AAPS PHARMSCITECH
·
2024
·Vol. 25
·期 6
·1-15
·
IF 3.4
Chen Jiasheng,Chen Zhenzhen,Wang Wentao,Wang Liyang,Zheng Jiaqi,Wu Shiqiong,Pan Yuru,Li Sai,Zhao Jie,Cai Zheng
Some glycoside drugs can be transported through intestinal glucose transporters (IGTs). The surfactants used in oral drug preparations can affect the function of transporter proteins. This study aimed to investigate the effect of commonly used surfactants, Poloxamer 188 and Tween 80, on the drug transport capacity of IGTs. Previous studies have shown that gastrodin is the optimal drug substrate for IGTs. Gastrodin was used as a probe drug to evaluate the effect of these two surfactants on intestinal absorption in SD rats through pharmacokinetic and in situ single-pass intestinal perfusion. Then, the effects of the two surfactants on the expression of glucose transporters and tight-junction proteins were examined using RT-PCR and western blotting. Additionally, the effect of surfactants on intestinal permeability was evaluated through hematoxylin-eosin staining. The results found that all experimental for Poloxamer 188 (0.5%, 2.0% and 8.0%) and Tween 80 (0.1% and 2.0%) were not significantly different from those of the blank group. However, the AUC (0-∞) of gastrodin increased by approximately 32% when 0.5% Tween 80 was used. The changes in IGT expression correlated with the intestinal absorption of gastrodin. A significant increase in the expression of IGTs was observed at 0.5% Tween 80. In conclusion, Poloxamer 188 had minimal effect on the drug transport capacity of IGTs within the recommended limits of use. However, the expression of IGTs increased in response to 0.5% Tween 80, which significantly enhanced the drug transport capacity of IGTs. However, 0.1% and 2.0% Tween 80 had no significant effect. Graphical Abstract
研究领域:
药剂学
·
药理学
·
肠道吸收
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产品关联
DOI
PubMed
高影响因子
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY
·
2019
·
IF 4.715
Qingquan Zeng,Mengru Bai,Cui Li,Shuanghui Lu,Zhiyuan Ma,Yunchun Zhao,Hui Zhou,Huidi Jiang,Dongli Sun,Caihong Zheng
Emtricitabine (FTC) is a first-line antiviral drug recommended for the treatment of AIDS during pregnancy. We hypothesized that transporters located in the placenta contribute to FTC transfer across the blood-placenta barrier. BeWo cells, cell models with stable or transient expression of transporter genes, primary human trophoblast cells (PHTCs), and small interfering RNAs (siRNAs) were applied to demonstrate which transporters were involved. FTC accumulation in BeWo cells was reduced markedly by inhibitors of equilibrative nucleoside transporters (ENTs), concentrative nucleoside transporters (CNTs), organic cation transporters (OCTs), and organic cation/carnitine transporter 1 (OCTN1) and increased by inhibitors of breast cancer resistance protein (BCRP) and multidrug resistance-associated proteins (MRPs). ENT1, CNT1, OCTN1, MRP1/2/3, and BCRP, but not ENT2, CNT3, OCTN2, or multidrug resistance protein 1 (MDR1), were found to transport FTC. FTC accumulation in PHTCs was decreased significantly by inhibitors of ENTs and OCTN1. These results suggest that ENT1, CNT1, and OCTN1 probably contribute to FTC uptake from maternal circulation to trophoblasts and that ENT1, CNT1, and MRP1 are likely involved in FTC transport between trophoblasts and fetal blood, whereas BCRP and MRP1/2/3 facilitate FTC transport from trophoblasts to maternal circulation. Coexistence of tenofovir or efavirenz with FTC in the cell medium did not influence FTC accumulation in BeWo cells or PHTCs.
研究领域:
药理学
·
妇产科学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
iScience
·
2024
·
IF 4.6
Yixi Su,Jiwei Chen,Jingyan Hu,Cheng Qian,Jiahao Ma,Sigurður Brynjolfsson,Weiqi Fu
A major obstacle to exploiting industrial flue gas for microalgae cultivation is the unfavorable acidic environment. We previously identified three upregulated genes in the low-pH-adapted model diatom Phaeodactylum tricornutum : ferredoxin (PtFDX), cation/proton antiporter (PtCPA), and HCO 3 − transporter (PtSCL4-2). Here, we individually overexpressed these genes in P. tricornutum to investigate their respective roles in resisting acidic stress (pH 5.0). The genetic modifications enabled positive growths of transgenic strains under acidic stress that completely inhibited the growth of the wild-type strain. Physiological studies indicated improved photosynthesis and reduced oxidative stress in the transgenic strains. Transcriptomes of the PtCPA- and PtSCL4-2-overexpressing transgenics showed widespread upregulation of various transmembrane transporters, which could help counteract excessive external protons. This work highlights ion/electron carrier genes’ role in enhancing diatom resistance to acidic stress, providing insights into phytoplankton adaptation to ocean acidification and a strategy for biological carbon capture and industrial flue gas CO 2 utilization.
研究领域:
植物学
·
遗传学
·
环境科学
·
生物技术
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2018
·Vol. 19
·期 5
·1394
·
IF 3.687
Danling Liang,Tianqiao Yong,Shaodan Chen,Yizhen Xie,Diling Chen,Xinxin Zhou,Dan Li,Muxia Li,Lu Su,Dan Zuo
Searching novel hypouricemic agents of high efficacy and safety has attracted a great attention. Previously, we reported the hypouricemic effect ofGanoderma applanatum, but its bioactives, was not referred. Herein, we report the hypouricemic effect of 2,5-dihydroxyacetophenone (DHAP), a compound screened fromGanoderma applanatumcomputationally. Serum parameters, such as uric acid (SUA), xanthine oxidase (XOD) activity, blood urea nitrogen (BUN), and creatinine were recorded. Real-time reverse transcription PCR (RT-PCR) and Western blot were exploited to assay RNA and protein expressions of organic anion transporter 1 (OAT1), glucose transporter 9 (GLUT9), uric acid transporter 1 (URAT1), and gastrointestinal concentrative nucleoside transporter 2 (CNT2). DHAP at 20, 40, and 80 mg/kg exerted excellent hypouricemic action on hyperuricemic mice, reducing SUA from hyperuricemic control (407 ± 31 μmol/L,p< 0.01) to 180 ± 29, 144 ± 13, and 139 ± 31 μmol/L, respectively. In contrast to the renal toxic allopurinol, DHAP showed some kidney-protective effects. Moreover, its suppression on XOD activity, in vivo and in vitro, suggested that XOD inhibition may be a mechanism for its hypouricemic effect. Given this, its binding mode to XOD was explored by molecular docking and revealed that three hydrogen bonds may play key roles in its binding and orientation. It upregulated OAT1 and downregulated GLUT9, URAT1, and CNT2 too. In summary, its hypouricemic effect may be mediated by regulation of XOD, OAT1, GLUT9, URAT1, and CNT2.Keywords:hyperuricemia;2,5-dihydroxyacetophenone;Ganoderma applanatum;bioactive compound
研究领域:
药理学
·
生物信息学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2019
·
IF 4.527
Fan Du,Peng Liu,Kai Wang,Zhaoguang Yang,Lin Wang
Uptake, translocation and speciation of As in rice plants have been investigated through hydroponic cultivations under stress from different As species. After germination, rice seedlings were treated with arsenite [As(III)], arsenate [As(V)], monomethylarsonic acid (MMA) or dimethylarsinic acid (DMA) at concentrations of 50, 100, 150, 200 and 250 μg/L for 24 days. Only inorganic As species were detected in the rice plants treated with As(III) or As(V), indicating that rice seedlings could not methylate inorganic As in hydroponic culture . As(V) in the rice roots was readily reduced to As(III) after uptake; thus, As(III) was the dominant species in the rice roots (>60%) and shoots (>80%) regardless of As(III) or As(V) treatment. The increased As(III) proportion in the nutrient solutions was due to the efflux of As(III) from the rice roots. MMA with relatively low stability in the blank nutrient solution was demethylated to As(III). Moreover, demethylation and methylation of MMA might occur simultaneously in rice plants. Specific proportions of MMA and AsB were observed in the rice roots treated with DMA, implying that MMA and AsB were the DMA metabolites in rice roots after detoxification.
研究领域:
植物学
·
环境科学
·
毒理学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）