JOURNAL OF MATERIALS CHEMISTRY
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2012
·Vol. 22
·期 23
·11543-11549
·
IF 5.968
Juan Qiao,Li Qi,Ying Shen,Lingzhi Zhao,Cui Qi,Dihua Shangguan,Lanqun Mao,Yi Chen
In this study, a novel fluorescent and temperature responsive block copolymer has been designed and synthesized by a reversible addition–fragmentation chain transfer (RAFT) polymerization method in terms of the strategy that N-isopropylacrylamide (NIPAm), maleic anhydride (MAn) and 7-amino-4-methylcoumarin (AMC) act as the temperature responsive unit, the hydrophilic unit and the fluorescent unit, respectively. The successfully synthesized block copolymer was characterized by gel permeation chromatography (GPC) and nuclear magnetic resonance (1H NMR) spectroscopy. Meanwhile, the self-aggregation behaviour in aqueous solution and the thermo-responsive property of the block copolymer were demonstrated by particle size measurement, transmission electron microscopy (TEM) observations and lower critical solution temperature (LCST) determination, respectively. Then the variation of fluorescence intensity with temperature was confirmed. With increasing temperature, shrinking of PNIPAm chains caused the block copolymer to become more hydrophobic above the LCST, assembling larger aggregates with lower interfacial curvature. Thus a part of the fluorescent groups would be embedded inside the enlarged block copolymer micelles, resulting in lower fluorescence intensity. Furthermore, the superior hydrophilicity and biocompatibility of the block copolymer as a thermometer have been demonstrated by application in intracellular temperature sensing of MDCK cells ranging from 24 °C to 38 °C.
研究领域:
材料科学
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生物医学工程
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高分子化学
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DOI
高影响因子
ACS Applied Materials & Interfaces
·
2017
·
IF 7.504
Feiyang Deng,Hua Zhang,Xing Wang,Yuan Zhang,Hongxiang Hu,Siyang Song,Wenbing Dai,Bing He,Ying Zheng,Xueqing Wang,Qiang Zhang
Drug nanocrystals (NCs) appear to be favorable to improving oral bioavailability of poorly water-soluble drugs as evidenced by the great success they have had in the market. However, the pathway and mechanism of drug NCs through epithelial membrane are still unclear. In an attempt to clarify their transport features, paclitaxel nanocrystals (PTX-NCs), and paclitaxel hybrid NCs with lipophilic carbocyanine dyes, were prepared and characterized as the models. The endocytosis, intracellular trafficking, paracellular transport, and transcytosis of PTX-NCs were carefully investigated with Förster resonance energy transfer (FRET) analysis, as well as a colocalization assay, flow cytometry, gene silencing, Western-blot, transepithelial electrical resistance (TEER) study and other approaches on MDCK cells. It was found that rod-like PTX-NCs could transport through the monolayer intact, and the process of endocytosis proved to be time and energy dependent. Endoplasmic reticulum (ER) and Golgi complexes were colocalized with PTX-NCs in cells, so the ER–Golgi complexes/Golgi complexes–basolateral membrane pathway may be involved in the intracellular trafficking and transcytosis of PTX-NCs. It was demonstrated here that cav-1, dynamin, and actin filament modulated the endocytosis process, and Cdc 42 regulated the transcytosis process. In addition, no paracellular transport of PTX-NCs was observed. These findings demonstrated that the rod-like nanocrystals not only enhanced the transcytosis of PTX compared with microparticles of raw drug materials but also changed the pathways of drug delivery. This study certainly provides insight for the oral absorption mechanism of nanocrystals of poorly soluble drugs.
研究领域:
药理学
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生物医学工程
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分子生物学
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产品关联
DOI
PubMed
高影响因子
CHEMISTRY & BIODIVERSITY
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2012
·Vol. 9
·期 9
·1916-1922
·
IF 1.804
Zhihan Xu,Chengyue Zhang,Yue Zhang,Xiaoda Yang
Measurement of paracellular permeation is an important assay for tight-junction investigations of drug toxicity, especially for metal-based drugs, and routine validation of the integrity of cell monolayers for models of drug absorption. Great efforts have been made in discovery and validation of novel paracellular diffusion indicators. In the present work, we prepared three Eu complexes, i.e. , [Eu(dtpa)] (dtpa=diethylenetriaminepentaacetic acid), [Eu(dtpa)(BSA)], and [Eu(dtpa)(PLL)] (PLL=poly( L -lysine)), and tested their permeation properties on Madin Darby canine kidney (MDCK) cells. The experimental results showed that all three probes were nontoxic to MDCK cells, permeated across MDCK monolayer exclusively via the paracellular pathways, and responded well to the changes on tight junction with high correlation of P app values to the decrease of trans-epithelial electric resistance (TEER). In addition, time-resolved fluorescence assays were conducted in a high-sensitivity and background-free mode. All these results confirmed the Eu complexes as novel and practical paracellular indicators.
研究领域:
药理学
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细胞生物学
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材料科学
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DOI
PubMed
高影响因子
RESEARCH IN VETERINARY SCIENCE
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2016
·
IF 1.504
Yong Sun,Tingting Guo,Dawei Guo,Li Guo,Li Chen,Yu Zhang,Liping Wang
Chicken P-glycoprotein (chP-gp), encoded by Abcb1 , determines the bioavailability because of its effect on pharmacokinetics of various drugs. However, comprehensive studies on chP-gp are still limited. In this study, the chicken full-length cDNA was first successfully cloned and then stably expressed in MDCK cell line. The open reading frame of chicken Abcb1 consists of 3864 nucleotides, encoding for a 1287-amino acid protein. Sequence alignments analysis showed that chicken P-gp had high identities with the homologues of turkey (95%), human (72%), pig (72%), rat (71%) and cattle (68%). The efflux ratio of rhodamine123 (Rho123, a human P-gp substrate) in ch Abcb1 transfected MDCK cells was significantly higher than that in the wild type MDCK cell (6.24 vs 1.64, P < 0.05), suggesting a good transporting function of chicken P-gp overexpressed in the transfected cell. Importantly, MDCK-ch Abcb1 cells, unlike Caco-2 cells, exhibited biphasic saturation kinetics in transporting Rho123. In conclusion, an MDCK cell line stably expressing ch Abcb1 was successfully established, which could provide a new cell model to screen its substrates and inhibitors and study the drug-drug interaction medicated via chicken P-gp.
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
产品关联
DOI
PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
·
2024
·
IF 4.8
Tongmei Shi,Jieling Lin,Shiyun Liang,Yu Song,Xin Zhao,Mengjie Xiao,Huihui Ti
A B S T R A C T Headings ethnopharmacological relevance Sangbaipi Decoction (SBPD) is an effective treatment for lung diseases caused by phlegm-heat obstruction according to Jingyue Quanshu , and soothes panting by purging the lung meridian. It is composed of anti-pyretic herbs (e.g., Scutellaria baicalensis Georgi and Coptis chinensis Franch.) and antitussive herbs (e.g., Cortex Mori and Armeniacae Semen Amarum ). Therefore, we hypothesized that SBPD has therapeutic effects on lung injury caused by influenza virus. Aim of the study This study aimed to explore anti-influenza activity, active components, and mechanisms of SBPD. Materials and methods The anti-influenza activities of SBPD were determined in 48 h drug-treated MDCK cell model using CPE and plaque reduction assays, and 24 h drug-treated A549 cells using qRT-PCR. The in vivo efficacy of SBPD (1.0g/kg/day and 0.5g/kg/day) was evaluated in PR8 infected BALB/c mice. The chemical component was assessed through HPLC-Q-TOF MS/MS analysis. Network pharmacology was built via TCMSP, GeneCards, DisgeNet, OMIM, DrugBank databases, and Cytoscape software. Additionally, TOA, HI and NAI assays were employed to investigate impact on the virus replication cycle with different concentrations of SBPD (2.5 mg/mL, 1.25 mg/mL, or 0.625 mg/mL). Results In MDCK infected with viruses A/PR/8/34, A/Hong Kong/1/68, or A/California/4/2009, the IC 50 values of SBPD were 0.80 mg/mL, 1.20 mg/mL, and 1.25 mg/mL. In A549 cells, SBPD treatment reduced cytokine expression (e.g., TNF-α, IL-6, IL-1β) ( p < 0.05). In PR8 infected BALB/c mice, SBPD improved the survival rate of infected mice, reduced lung index ( p < 0.05), protected lung tissue from pathological damage, and regulated cytokine overexpression ( p < 0.05). 29 components of SBPD were identified in SBPD treated mouse serum including some phytochemicals targeting influenza proteins. HI and NAI assays suggested the potential antiviral mechanism of SBPD through inhibition of HA and NA. Conclusion This study is the first to demonstrate the anti-influenza and the anti-inflammatory effects of SBPD in vitro and in vivo. Its major anti-influenza phytochemicals were explored and its inhibitory effects on HA and NA protein were proved. It provides more options for anti-influenza drug discovery.
研究领域:
药理学
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病毒学
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中医
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免疫学
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产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2025
·Vol. 15
·期 1
·1-14
·
IF 3.9
Zhou Lei,Zhang Mengting,Sun Huinan,Duan Liyuan,Meng Qingwei
This study establishes the first reversed-phase HPLC-UV method for simultaneous baseline separation of 12 structurally diverse water-soluble vitamins (WSVs) within 25 min. The eco-friendly approach utilizes a methanol–potassium dihydrogen phosphate (pH 4.85) gradient on a ChromCore C18 column (250 × 4.6 mm, 5 μm) with dual-wavelength detection (210/266 nm), eliminating environmentally detrimental ion-pairing reagents. Rigorous validation per ICH Q2(R1) guidelines confirmed excellent linearity (R² > 0.999), precision (RSD < 2.1%), accuracy (spike recoveries: 93.9–106.8%), and sensitivity (LODs: 0.01–0.78 µg/mL). The method resolved key analytical challenges including polarity disparities, matrix interference, and vitamin instability. Successful quantification of endogenous vitamins in DMEM medium showed ≤ 5.1% deviation from certified values for 11/12 analytes. Applied to five commercial MDCK cell culture media, the method revealed significant inter-product variability in vitamin composition – notably universal under-supplementation of riboflavin (0.576–0.741 µg/mL, 10× lower than other vitamins) and inconsistent biotin/adenine supplementation. This robust, cost-effective platform provides biomanufacturers with a powerful quality control tool for optimizing vitamin supplementation in vaccine production systems, addressing a critical gap in cell culture analytics.
研究领域:
分析化学
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生物化学
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生物技术
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药学
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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 PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2023
·
IF 3.4
Jieyu Zhang,Yan Chen,Wenxiang Fan,Linnan Li,Yueming Ma,Zhengtao Wang,Rong Shi,Li Yang
The combined efficacy in lowering serum lipid levels and increasing kidney protection of Plantago asiatica L. seed (Plantago) and Coptis chinensis Franch. rhizoma (Coptis) is far better than the effects of either herb alone. This finding suggests that there must be some degree of herb-herb interactions (HHI) affect potency. Here, we chose geniposidic acid (GPA), acteoside (ACT), and plantagoamidinic acid A (PLA) as active components in Plantago, and berberine (BBR) as the active component in Coptis, and, using transporter gene-transfected Madin-Darby canine kidney (MDCK) cells in combination with specific substrates and inhibitors, investigated Plantago- Coptis HHIs. We also established a UPLC-MS/MS analytical method to determine substrate content. Results showed that PLA in Plantago was a substrate of rOCT1/2 and rMATE1, and had inhibitory effects on rOCT2 and rMATE1. We also found that ACT is a substrate of rMATE1, but GPA was not a substrate of any transporter that we investigated. When BBR was used as the substrate, the inhibition rate of 10 μM PLA was 53.6% on rOCT2 and 31.5% on rMATE1. The inhibition rates of 30 μM ACT and 30 μM GPA on rMATE1 were 47.0% and 31.0%, respectively. Thus, our findings suggest that GPA, ACT, PLA, and BBR have competitive interactions that are driven by the rOCT2 and rMATE1 transporters. These interactions affect the transport and excretion of compounds and result in efficacy changes after co-administration.
研究领域:
药理学
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药学
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中药学
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分析化学
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产品关联
DOI
PubMed
高影响因子
International Journal of Nanomedicine
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2020
·
IF 5.115
Jing-Yi Ye,Zhong-Yun Chen,Chuan-Li Huang,Bei Huang,Yu-Rong Zheng,Ying-Feng Zhang,Ban-Yi Lu,Lin He,Chang-Shun Liu,Xiao-Ying Long
Objective A non-lipolysis nanoemulsion (NNE) was designed to reduce the first-pass metabolism of raloxifene (RAL) by intestinal UDP-glucuronosyltransferases (UGTs) for increasing the oral absorption of RAL, coupled with in vitro and in vivo studies. Methods In vitro stability of NNE was evaluated by lipolysis and the UGT metabolism system. The oral bioavailability of NNE was studied in rats and pigs. Finally, the absorption mechanisms of NNE were investigated by in situ single-pass intestinal perfusion (SPIP) in rats, Madin-Darby canine kidney (MDCK) cells model, and lymphatic blocking model. Results The pre-NNE consisted of isopropyl palmitate, linoleic acid, Cremophor RH40, and ethanol in a weight ratio of 3.33:1.67:3:2. Compared to lipolysis nanoemulsion of RAL (RAL-LNE), the RAL-NNE was more stable in in vitro gastrointestinal buffers, lipolysis, and UGT metabolism system ( p < 0.05). The oral bioavailability was significantly improved by the NNE (203.30%) and the LNE (205.89%) relative to the suspension group in rats. However, 541.28% relative bioavailability was achieved in pigs after oral NNE intake compared to the suspension and had two-fold greater bioavailability than the LNE ( p < 0.05). The RAL-NNE was mainly absorbed in the jejunum and had high permeability at the intestine of rats. The results of both SPIP and MDCK cell models demonstrated that the RAL-NNE was absorbed via endocytosis mediated by caveolin and clathrin. The other absorption route, the lymphatic transport (cycloheximide as blocking agent), was significantly improved by the NNE compared with the LNE ( p < 0.05). Conclusion A NNE was successfully developed to reduce the first-pass metabolism of RAL in the intestine and enhance its lymphatic transport, thereby improving the oral bioavailability. Altogether, NNE is a promising carrier for the oral delivery of drugs with significant first-pass metabolism.
研究领域:
药剂学
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药物动力学
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药物递送系统
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药学
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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
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Viruses-Basel
·
2024
·Vol. 16
·期 5
·768
·
IF 3.8
Sixu Liu,Jingqi Li,Qingtian Cheng,Kangyi Duan,Zhan Wang,Shuang Yan,Shuaishuai Tian,Hairui Wang,Shaobin Wu,Xinkui Lei,Yu Yang,Ningning Ma
Influenza vaccines, which are recommended by the World Health Organization (WHO), are the most effective preventive measure against influenza virus infection. Madin–Darby canine kidney (MDCK) cell culture is an emerging technology used to produce influenza vaccines. One challenge when purifying influenza vaccines using this cell culture system is to efficiently remove impurities, especially host cell double-stranded DNA (dsDNA) and host cell proteins (HCPs), for safety assurance. In this study, we optimized ion-exchange chromatography methods to harvest influenza viruses from an MDCK cell culture broth, the first step in influenza vaccine purification. Bind/elute was chosen as the mode of operation for simplicity. The anion-exchange Q chromatography method was able to efficiently remove dsDNA and HCPs, but the recovery rate for influenza viruses was low. However, the cation-exchange SP process was able to simultaneously achieve high dsDNA and HCP removal and high influenza virus recovery. For the SP process to work, the clarified cell culture broth needed to be diluted to reduce its ionic strength, and the optimal dilution rate was determined to be 1:2 with purified water. The SP process yielded a virus recovery rate exceeding 90%, as measured using a hemagglutination units (HAUs) assay, with removal efficiencies over 97% for HCPs and over 99% for dsDNA. Furthermore, the general applicability of the SP chromatography method was demonstrated with seven strains of influenza viruses recommended for seasonal influenza vaccine production, including H1N1, H3N2, B (Victoria), and B (Yamagata) strains, indicating that the SP process could be utilized as a platform process. The SP process developed in this study showed four advantages: (1) simple operation, (2) a high recovery rate for influenza viruses, (3) a high removal rate for major impurities, and (4) general applicability.
研究领域:
生物技术
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病毒学
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药学
✓ 已验证
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）