Hollow Fiber Cell Fishing Research Articles

Hollow fibre cell fishing and hollow fibre liquid phase microextraction research on the anticancer coumarins of Radix Angelicae dahuricae in vitro and in vivo

Hollow fiber cell fishing (HFCF) and hollow fiber liquid phase microextraction (HF-LPME) with high performance liquid chromatography were utilized to research the anti-hepatoma HepG-2 or anti-renal tubular ACHN cells coumarins from Radix Angelicae dahuricae (RAD) in vitro and vivo. Before application, the coumarins of psoralen, bergapten, oxypeucedanin, imperatorin and isoimperatorin were selected as model compounds, the experiment conditions and methodology parameters of both HFCF and HF-LPME were investigated. Under the optimized conditions, cell growth state and survival rate, cell specific binding, repeatability for HFCF, and linear ranges, limits of quantification (1.3–21.0 ng/mL), precisions (<15.2%), recoveries (80.4%–109.3%) and stability for HF-LPME were achieved. Results showed that the five model compounds and xanthotoxol, xanthotoxin were screened in RAD extraction; xanthotoxin and xanthotoxol were fished by HepG-2 cell from both plasma and liver, xanthotoxol, xanthotoxin, bergapten and imperatorin were fished by ACHN cell from plasma and kidney; and these coumarins maintained a certain concentration in vivo. The study results provide experimental basis for researching the active anticancer components and expound the medicinal material base in RAD. The combination of HFCF and HF-LPME is simple and effective in studying bioactive anticancer compounds in traditional Chinese medicine in vitro and in vivo.

Research on major antitumor active components in Zi-Cao-Cheng-Qi decoction based on hollow fiber cell fishing with high performance liquid chromatography

Hollow fiber cell fishing (HFCF) based on hepatoma HepG-2 cells, human renal tubular ACHN cells or human cervical carcinoma HeLa cells, coupled with high-performance liquid chromatography (HPLC), was developed and employed to research the major active components in Zi-Cao-Cheng-Qi decoction both in vitro and in vivo. The research showed that the active components, such as hesperidin, magnolol, honokiol, shikonin, emodin and β,β′-dimethylacrylshikonin were screened out by HFCF based on the cancer cells in vitro, furthermore they can be absorbed into blood and reach in the target organ, and some of the active components can be fished by the cells and maintain effective concentrations. Before application of HFCF with HPLC, cell growth state, cell survival rate, positive effect on screening results binding between active centers on the fiber and target components, repeatability of retention times and relative peak areas of the target analytes were analysed and investigated. In short, HFCF with HPLC is a simple, inexpensive, effective, and reliable method that can be used in researching active components from traditional Chinese medicine (TCM) and its formula both in vitro and in vivo, elucidating preliminarily the TCM characteristics of multiple components and multiple targets, laying a foundation for expounding the antitumor efficacy material basis in TCM.

Study of Anti-Renal Cancer Ingredients in Scutellaria barbata on Hollow Fibre Cell Fishing and Hollow Fibre Liquid Phase Microextraction

Study of Anti-Renal Cancer Ingredients in Scutellaria barbata on Hollow Fibre Cell Fishing and Hollow Fibre Liquid Phase Microextraction

Study on major antitumor components in Yinchenhao decoction in vitro and in vivo based on hollow fiber cell fishing coupled with high performance liquid chromatography.

Hollow fiber cell fishing (HFCF) based on human hepatoma cell HepG-2 or human renal tubular cell ACHN coupled with high performance liquid chromatography/ultraviolet detection (HPLC/UV) was developed and used for simultaneous study of the major antitumor active components in a formula of Yinchenhao decoction (YCHD) in vitro and in vivo, and in its constituent herbs, namely, Artemisia capillaris Thunb, Gardenia jasminoides Ellis, Radix et Rhizoma Rhei in vitro. Before application, chlorogenic acid, geniposide, p-hydroxyacetophenone, crocin and rhein were chosen as model compounds, the various validations, e.g., cell growth and cell viability on the fiber inner wall, binding between fiber active center and component, repeatability of retention time or relative peak area of the active components were investigated. We screened and identified the major antitumor active components of YCHD, verified their synergetic or antagonistic effect, correlated the major active components between in vitro and in vivo and determined major effective components. Our study will serve as a valuable reference in probing the antitumor material basis of YCHD and its three constituent herbs, as well as in identifying novel clinical aspects of traditional Chinese medicines. The results showed that HCFC-HPLC is a simple, rapid, and reliable method that simultaneously researches the interaction between multiple targets and multiple components of herbal medicine.

Study on Antitumour Activity of Scutellarin and Its Metabolite Scutellarein by Combining Activity Screening, Target Tissue Distribution and Pharmacokinetics

Ingredients in traditional Chinese medicine (TCM) can be absorbed into blood and reach and maintain effective concentrations in the target organ, which is the premise of the ingredients taking effect in vivo. Hollow fibre cell fishing (HFCF) based on ACHN human renal tubular cells or HepG2 hepatoma cells using high-performance liquid chromatography with ultraviolet detection (HPLC/UV), coupled with hollow fibre liquid-phase microextraction (HF-LPME) with HPLC/UV, was developed and employed for the first time to study the antitumour activity of scutellarin, a main ingredient in breviscapine, and its metabolite scutellarein after intraperitoneal injection of breviscapine. The antitumour activity of these ingredients was evaluated by combining their activity screening, target tissue distribution and pharmacokinetics (PK). Before application, the cell growth state, survival rate, binding between fibre active centre or nutrient solution and ingredients, positive and negative control and repeatability for HFCF were investigated; validations of the HF-LPME methodology were optimized and evaluated. The study found that scutellarin can be rapidly absorbed into blood and metabolize partly into scutellarein, and both can reach liver or kidney, combine with ACHN or HepG2 cells and maintain effective concentrations. Thus, scutellarin and scutellarein have potential as lead ingredients for treatment of liver or kidney tumours, and breviscapine injection may have potential properties for use in treatment or prevention of liver and kidney tumours, at least. HFCF and HF-LPME coupled with HPLC can contribute to simultaneous investigation of the antitumour activity of multiple ingredients in TCMs by combining activity screening, tissue distribution and PK studies.

Screening and quantification of anticancer compounds in traditional Chinese medicine by hollow fiber cell fishing and hollow fiber liquid/solid-phase microextraction.

Hollow fiber cell fishing, based on HepG-2, SKOV-3, and ACHN cancer cells, and hollow fiber liquid/solid microextraction with HPLC were developed and introduced for researching the anticancer activity of Rhizoma Curcumae Longae, Radix Curcumae, and Rhizoma Curcumae. The structures of curcumin, demethoxycurcumin, and bisdemethoxycurcumin screened were identified and their contents were determined. The compound target fishing factors and cell apoptosis rates under the effect of the three medicines were determined. The binding sites (cell membrane and cell organelle) and binding target (phospholipase C) on the cell were researched. Hollow fiber liquid/solid-phase microextraction mechanism was analyzed and expounded. Before the application, cell seeding time, growth state and survival rate, compound nonspecific binding, positive and negative controls, repeatability in hollow fiber cell fishing with high-performance liquid chromatography; extraction solvent, sample pH, salt concentration, agitation speed, extraction time, temperature and sample volume in hollow fiber liquid/solid-phase microextraction with high-performance liquid chromatography were investigated. The results demonstrated that the proposed strategy is a simple and quick method to identify bioactive compounds at the cellular level as well as determine their contents (particularly trace levels of the bioactive compounds), analyze multicompound and multitarget entirety effects, and elucidate the efficacious material base in traditional medicine.

Screening and Research of Anti-Cancer Matrine Components Based on Hollow Fiber Cell Fishing with High-Performance Liquid Chromatography

Hollow fiber cell fishing with high-performance liquid chromatography (HFCF-HPLC) based on human breast cancer MCF-7 cells, human renal tubular ACHN cells, human ovarian cancer SKOV-3 cells, and hepatoma HepG-2 cells, and their cell membranes, cell organelles, or receptors was developed and employed to research the active matrine alkaloids group, such as oxymatrine, matrine, sophoridine, and oxysophocarpine, from radix sophorae flavescentis (RSF) and its injection in vitro. Hollow fiber liquid-phase microextraction with HPLC (HF-LPME-HPLC) was conducted to extract and detect the active components in vivo. Some of the structures of the active components screened were identified in vitro. The binding sites and targets of the potential active matrine compounds were preliminarily researched. The effective anti-cancer components were determined, and the efficacy material basis of both medicines on matrine alkaloids was elucidated. Before the application of HFCF-HPLC, the survival rate of cells seeded on the fiber, death rate after screening components, non-specific binding between active centers on the fiber and target components, positive and negative controls, and repeatability of the tests were investigated. Results demonstrated that HFCF-HPLC, coupled with HF-LPME-HPLC, is a simple, quick, and reliable method to screen active components, determine truly effective components, and elucidate preliminarily the characteristics of multiple components and multiple targets for traditional Chinese medicine.

Research of Anti-Cancer Components in Traditional Chinese Medicine on Hollow Fibre Cell Fishing and Hollow Fibre Liquid Phase Microextraction

Hollow fibre cell fishing with HPLC (HFCF-HPLC) based on the human ovarian cancer cell line SKOV-3, human renal tubular cell line ACHN or hepatoma cell line HepG-2 was employed to screen active groups of coumarin and volatile oil in Radix angelicae sinensis, Radix angelicae dahuricae and Fructus citri sarcodactylis. Simultaneously, hollow fibre liquid phase microextraction with HPLC (HFLPME-HPLC) was conducted to enrich and determine the contents of active components in the same sample solution. Before application, for HFCF-HPLC, cells growth states and survival rates on the fibre, effect of ethanol concentration in the extract of samples on cell survival rates, non-specific binding between fibre active centres and the target components, positive and negative controls and repeatabilities were validated; for HFLPME, extraction solvent, sample phase pH, agitation speed, extraction time and sample phase volume were investigated. Many active components were screened from three medicines. Some of them, such as scoparone, psoralen, bergapten, oxypeucedanin, imperatorin, ligustilide, were identified by MS. The target fishing factors of active components and the cell apoptosis rates of three cells under the medicines effect were researched. The binding sites of active groups on HepG-2 cells were preliminarily determined. The results demonstrated that HFCF-HPLC, coupled with HFLPME-HPLC, is a simple and universal approach to find bioactive components at the cellular level, determine their content and research traditional Chinese medicines (TCMs) entirety effect of multi-component and multi-target. The approach may provide us a new and good solution to clarify the material basis of anti-cancer effect and conduct personalized quality control for the components associated with efficacy in TCMs.

Rapid screening of different types of antitumor compound groups from traditional chinese medicine by hollow fiber cell fishing with high performance liquid chromatography.

A novel hollow fiber cell fishing with high performance liquid chromatography (HFCF-HPLC) was extended and used to screen flavonoid and anthraquinone active compound groups simultaneously from traditional Chinese medicines (TCMs). In this study, three cells (MCF-7, SGC7901, and MADB-106) were seeded on the inner wall of the hollow fiber employed to screen bioactive components from TCM water decoction. The variables influencing HFCFHPLC, such as cell seeding time, screening stirring rate and time, and active compound concentration, were investigated and optimized. The surface property of the hollow fiber seeded with cells, the cell survival rate under different conditions, the nonspecific binding between active centers in the fiber and the target compounds, and the repeatability and recovery of HFCF-HPLC were analyzed and validated. Certain structures of the compounds fished by HFCF-HPLC were identified after comparing the retention times of the reference substances. To verify preliminarily the binding site between the bioactive components and cells, we separated the cell membrane and cell organelle from live MCF-7 cells. We then employed the cell membrane, cell organelle, and the whole cells to screen simultaneously the active compounds. The cell fishing factor of the active compound was calculated and discussed as the index of cell-drug binding ability in HFCFHPLC. Tamoxifen as a positive control and indomethacin as a negative control were screened by HFCF-HPLC to verify the method. The results indicate that HFCF-HPLC is an effective and reliable method for the screening and analysis of bioactive components. Moreover, this method can be applied to predict bioactive candidates in TCMs.

Screening of bioactive compounds and research of possible targets based on hollow fiber cell fishing with high performance liquid chromatography

In this study, hollow fiber cell fishing with high performance liquid chromatography (HFCF-HPLC) is further extended and verified. The surface properties of the hollow fiber seeded human breast cancer cell MCF-7, the survival rate of cells seeded, the cell death rate after screening of active compounds, the non-specific binding between active centres on the fiber and the target compounds, the positive and negative controls and the repeatability of HFCF-HPLC were investigated in detail. The target fishing factor of active compounds was defined and improved. The procedure was employed to screen and fish simultaneously phenolic acid and flavonoid active compound groups from water decoctions of traditional Chinese medicines (TCMs). Some of the structures of active compounds screened and fished were identified by comparing with the retention time of the reference substances. And the hollow fibers filled with the receptors (estrogen receptor, epidermal growth factor receptor, and cyclooxygenase-2) associated with cancer cells as well as the cell membranes and cell organelles separated from live MCF-7 cells were used respectively in preliminary research of the binding sites and the targets of potential phenolic acids and flavonoids on MCF-7 cells. The active compounds' abilities to interact with their targets were compared and are discussed. Results demonstrate that HFCF-HPLC is a simple, quick, stable and reliable method to screen and analyze bioactive compounds as well as research the possible targets of active compounds, and it provides us with a good possibility to research and prove the effects of multiple components and multiple targets for TCMs.

Hollow fiber cell fishing with high-performance liquid chromatography for rapid screening and analysis of an antitumor-active protoberberine alkaloid group from Coptis chinensis

A novel hollow fiber cell fishing procedure with high-performance liquid chromatography (HFCF–HPLC) was developed and used for rapid screening, fishing, and analysis of bioactive compounds from traditional Chinese medicines. Human breast cancer cell MCF7, mouse breast cancer cell MADB106, and gastric cancer cell SGC7901 were seeded on the internal surface of hollow fibers that were used to screen, fish, and analyze an antitumor-active protoberberine alkaloid group from Coptis chinensis decoction. The main variables that affect the HFCF–HPLC process were investigated and optimized. The surface properties of the hollow fiber-seeded cells, cell survival rate, non-specific binding between active centers in the hollow fiber and the target compounds, repeatability, reliability, and recovery of HFCF–HPLC were investigated in detail. Several active compounds structures that were screened from Coptis chinensis by using HFCF–HPLC were identified by comparing the retention time of the reference substances. The cell membrane and cell organelle were separated from MCF7 cells for a preliminary study of the target effect of active compounds on MCF7 cells. The living cell, cell membrane, and cell organelle fishing factors of the active compound, as the indexes of drug binding ability in HFCF–HPLC, were defined and discussed. In addition, tamoxifen as positive control substance and indomethacin as negative control substance were screened by using HFCF–HPLC to further verify the method's reliability. The results demonstrated that HFCF–HPLC is an effective, rapid, stable, and reliable method to screen and analyze bioactive compounds.

Hollow fibre cell fishing with high performance liquid chromatography for screening bioactive anthraquinones from traditional Chinese medicines

Hollow fibre cell fishing with high performance liquid chromatography (HFCF-HPLC) is a newly developed method used to screen and fish bioactive compounds in traditional Chinese medicines (TCMs). In the study, colorectal cancer cell HCT116 was first seeded in a hollow fibre and used for screening and fishing active compounds from TCMs. The surface properties of the hollow fibre seeded with HCT116 cells, the non-specific binding between active centres in the fibre and the target compounds, the cell survival rate under different conditions before and after screening, the repeatability and recovery of HFCF-HPLC were investigated in detail. The cell fishing factor of active compound was defined in HFCF-HPLC. We employed HFCF-HPLC to screen and fish anthraquinones active compounds group from extracts of Polygonum cuspidatum, Cecropia obtusifolia L. and Polygoni multiflori radix praeparata. Some of the anthraquinones structures screened from TCMs were identified by comparing to the retention time of the reference substances confirmed by mass spectrometry. The ability of permeable membrane of anthraquinones screened by HFCF-HPLC was further described. Indomethacin was used as the positive control substance. Results demonstrated that HFCF-HPLC is an effective, stable and reliable method to screen and analyse bioactive compounds. Other bioactive compounds from TCMs could also be screened.

Hollow fiber cell fishing with high performance liquid chromatography for screening bioactive compounds from traditional Chinese medicines

A novel hollow fiber cell fishing method with high performance liquid chromatography was proposed and used to screen, isolate, and analyze bioactive compounds from Traditional Chinese Medicines (TCMs). The active compounds that interact with the living cells acceptor inside the hollow fiber lumen were screened and isolated from the TCM extracts in phosphate buffer solution (pH 7.4). Subsequently, the active compounds bound to the cells were desorbed with methanol, and were analyzed using HPLC. HFCF with HPLC was introduced for the screening and analysis of lignans in Schisandra chinensis (Turcz) Baill and coumarins in Fructus Cnidii and Fructus Psoraleae. The surface properties of the hollow fibers filled with living cells were characterized. The nonspecific binding between the active centers of the hollow fibers and the bioactive compounds were investigated. The cell survival rates were determined before and after the screening. The repeatability of the method was tested. Some structures of the lignans and coumarins screened from TCMs were identified by the comparison to the retention times of the reference substances. HFCF-HPLC is a simple, fast, effective, and reliable method for the screening and analysis of bioactive compounds, and it can be extended to screen other bioactive compounds from TCMs.