CHEMICO-BIOLOGICAL INTERACTIONS
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2018
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IF 3.296
Shishi Li,Jinjin Xu,Zhihong Yao,Liufang Hu,Zifei Qin,Hao Gao,Kristopher W. Krausz,Frank J. Gonzalez,Xinsheng Yao
Cycloicaritin is a bioactive natural phenolic compound from Epimedium species. However, the glucuronidation and excretion which would influence oral bioavailability and pharmacokinetics of cycloicaritin still remain unknown. Here we aimed to establish UGT1A1 stably transfected HeLa cells, and to determine the contributions of BCRP and MRPs transporters to excretion of cycloicaritin-3- O -glucuronide. First, β-estradiol was used to validate the expression of active UGT1A1 protein in engineered HeLa1A1 cells. Furthermore, Ko143 (5 and 20 μM) led to a significant decrease (42.4%–63.8%, p < 0.01) in CICT-3-G excretion and obvious accumulation (19.7%–54.2%, p < 0.05) of intracellular CICT-3-G, while MK571 (5 and 20 μM) caused a significant reduction (46.8%–64.8%, p < 0.05) in the excretion and obvious elevation (50.7%–85.2%, p < 0.01) of intracellular level of CICT-3-G. Furthermore, BCRP knocked-down brought marked reduction in excretion rates of CICT-3-G (26.0%–42.2%, p < 0.01), whereas MRP1 and MRP4-mediated silencing led to significant decrease in the excretion of CICT-3-G (23.8%–35.4%, p < 0.05 for MRP1 and 11.9%–16.0%, p < 0.05 for MRP4). By contrast, neither CICT-3-G excretion nor CICT-3-G accumulation altered in MRP3 knocked-down cells as compared to scramble cells. Taken together, BCRP, MRP1 and MRP4 were identified as the most important contributors for CICT-3-G excretion. Meanwhile, the UGT1A1 modified HeLa cells were a simple and practical tool to study UGT1A1-mediated glucuronidation and to characterize BCRP and MRPs-mediated glucuronide transport at a cellular level.
研究领域:
药理学
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分子生物学
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药物代谢
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MOLECULAR PHARMACEUTICS
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2015
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IF 4.384
Xingwang Zhang,Dong Dong,Huailing Wang,Zhiguo Ma,Yifei Wang,Baojian Wu
Efflux of glucuronide is facilitated by the membrane transporters including BCRP and MRPs. In this study, we aimed to determine the effects of transporter expression on glucuronide efflux and cellular glucuronidation. Single efflux transporter (i.e., BCRP, MRP1, MRP3, or MRP4) was stably knocked-down in UGT1A1-overexpressing HeLa cells. Knock-down of transporters was performed by stable transfection of short-hairpin RNA (shRNA) using lentiviral vectors. Glucuronidation and glucuronide transport in the cells were characterized using three different aglycones (i.e., genistein, apigenin, and emodin) with distinct metabolic activities. BCRP knock-down resulted in significant reductions in excretion of glucuronides (42.9% for genistein glucuronide (GG), 21.1% for apigenin glucuronide (AG) , and 33.7% for emodin glucuronide (EG); p < 0.01) and in cellular glucuronidation (38.3% for genistein, 38.6% for apigenin, and 34.7% for emodin; p < 0.01). Knock-down of a MRP transporter led to substantial decreases in excretion of GG (32.3% for MRP1, 36.7% for MRP3, and 36.6% for MRP4; p < 0.01) and AG (59.3% for MRP1, 24.7% for MRP3, and 34.1% for MRP4; p < 0.01). Also, cellular glucuronidation of genistein (38.3% for MRP1, 32.3% for MRP3, and 31.1% for MRP4; p < 0.01) and apigenin (40.6% for MRP1, 32.4% for MRP3, and 34.6% for MRP4; p < 0.001) was markedly suppressed. By contrast, silencing of MRPs did not cause any changes in either excretion of EG or cellular glucuronidation of emodin. In conclusion, cellular glucuronidation was significantly altered by decreasing expression of efflux transporters, revealing a strong interplay of glucuronidation with efflux transport.
研究领域:
药理学
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分子生物学
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细胞生物学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2024
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IF 6.7
Lihong Yu,Chenglin Yang,Shuting Cheng,Qianqian Jiang,Yuehong Pang,Xiaofang Shen
The recognition of small molecules plays a crucial role in disease diagnosis, environmental assessment, and food safety. Currently, their recognition elements predominantly rely on antibodies and aptamers while suffering from a limitation of the complex screening process due to the low immunogenicity of small molecules. Herein, we present a top-down computational design strategy for molecule recognition peptides (MRPs) for enzyme-peptide self-assembly and chemiluminescence biosensing. Taking ochratoxin A (OTA) as an illustrative example, human serum albumin (HSA) was selected as the parental protein due to its high affinity for OTA binding. Through iterative computational simulations involving the binding domain of the HSA-OTA complex, our strategy identified a specific 15-mer MRP (RLKCASLKFGERAFK), which possesses excellent binding affinity (38.02 ± 1.24 nM) against OTA. Molecular dynamics simulations revealed that the 15-mer MRP unfolds into a flexible short chain with high affinity for OTA, but exhibits weak or no binding affinity with five structurally similar mycotoxins. Furthermore, we developed a novel enzyme-peptide self-assembly approach mediated by calcium(II) to obtain nanoflowers, which integrates both the recognition element (MRP) and the signal translator (enzyme) for chemiluminescence biosensing. The assembled nanoflowers allow MRPs to be directly utilized as a tracer for OTA biosensing without labeling or secondary antibodies. This computational-to-application approach offers a new route for small-molecule recognition.
研究领域:
生物化学
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计算生物学
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分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF PHARMACOLOGY
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2015
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IF 2.532
Pan Chen,Jingjie Li,Xiaomei Fan,Hang Zeng,Rongrong Deng,Dongshun Li,Min Huang,Huichang Bi
Obstructive cholestasis is characterized by impairment of hepatic canalicular bile efflux and there are no clinically effective drugs to cure except surgeries. Previously we revealed that oleanolic acid (OA) protected against lithocholic acid (LCA)-induced intrahepatic cholestasis in mice. Cholestasis caused by LCA is characterized by segmental bile duct obstruction, whether OA possesses the beneficial effect on completed obstructive cholestasis induced by bile duct ligation (BDL) remains unknown. In this study, we demonstrated that BDL-induced mice liver pathological change, and increase in serum levels of ALT, AST and ALP were all significantly reduced by OA (20 mg/kg, i.p.). Meanwhile, OA also lowered total bilirubin and total bile acids levels in serum, as well as total bile acids level in liver, in contrast, urinary total bile acids output was remarkably up-regulated by OA. Gene expression analysis showed that OA caused significant increased mRNA expression of MRP3 and MRP4 located at hepatic basolateral membrane , and restoration of MRP2 and BSEP located at hepatic cannalicular membrane. Furthermore, significant NRF2 protein accumulation in nucleus was also observed in OA treated mice. In mice primary cultured hepatocytes, the effects of OA on MRP2, MRP3 and MRP4 expression were directly proved to be mediated via NRF2 activation, and BSEP downregulation induced by OA was in part due to FXR antagonism. Luciferase assay performed in Hep G2 cells also illustrated that OA was a partial FXR antagonist. Taken together, we conclude that OA attenuates obstructive cholestasis in BDL mice, possibly via activation of NRF2-MRPs and FXR antagonism.
研究领域:
肝病学
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药理学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Wen-mei Chen,Yang Wang,Xu-mei Wang,Yan-hong Shao,Zong-cai Tu,Jun Liu
This study analyzes the interactions among Maillard reaction products (MRPs), digestibility and antioxidant activity in a β-lactoglobulin-glucose (βlg-Glu) model system during superheated steam treatment at 120 °C for 1 to 5 min. With an increase in treatment duration, there were significant increases in glucose loss, grafting degree, browning, and the formation of intermediate products in βlg-Glu. Characteristic MRPs, including α-dicarbonyl compounds, 5-hydroxymethylfurfural, and advanced glycation end products (AGEs), were formed through the degradation of sugars and condensation reactions between carbonyls and amines, accompanied by an increase in oxidative products. These changes impacted the molecular weight distribution and conformational structure of βlg-Glu, resulting in decreased digestibility. βLg-Glu with the highest level of glycation exhibited superior antioxidant activity after in vitro digestion, which was closely associated with the increase in AGEs and oxidation products. Therefore, the protein-sugar system treated by superheated steam with different heating time can significantly affect the formation and activity of the products, providing theoretical basis for superheated steam processing nutritious and healthy food.
研究领域:
食品科学
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营养学
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生物化学
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食品加工
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL DAIRY JOURNAL
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2023
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IF 3.1
Hongbo Li,Meng Bian,Yuelu Jiang,Chunshuang Li,Yang Zou,Xiaohong Chen,Hongjuan Li,Jinghua Yu
To better monitor and reduce the generation of Maillard reaction products (MRPs) during the heat treatment and storage of dairy products , a reaction kinetic model was established to evaluate the inhibitory effects of tea polyphenols on MRPs in the milk simulation system. The inhibition of MRPs by tea polyphenols followed first order kinetics, while the inhibition of 5-hydroxymethyl-2-furfural (5-HMF) was most pronounced at 95 °C with the reaction rate constant of −0.0683 d −1 . Arrhenius model showed that the conversion rate of 5-HMF (35.672 d −1 ) was higher than N ε -carboxyethyllysine (2.585 d −1 ) and N ε -carboxymethyllysine (0.53 d −1 ) after the addition of tea polyphenols, which indicated that 5-HMF was the main intermediate product of Maillard reaction. Moreover, the shelf life of the two-stage sterilised milk containing tea polyphenols was 141 d, which was 2.1 times longer than that of the control sample (67 d), which provided reference for consumers during the drinking period.
研究领域:
食品科学
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化学动力学
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乳品技术
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食品保鲜
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）