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安徽医科大学学报 2008年第4期43卷 398-402页

作者：许伟张循善钟涛王明丽安徽医科大学第一附属医院输血科合肥230022 安徽医科大学微生物学教研室合肥230032

目的探讨核黄素光化学法灭活单采血小板细菌的效果及血小板一系列生化和生理学指标的改变。方法将28ml浓度为500μmol／L核黄素加到250ml的单采血小板悬液中，再将一定浓度的G^＋菌（表皮葡萄球菌ATCC12228）和G^＋菌（大肠埃希菌ATCC25... 详细信息

目的探讨核黄素光化学法灭活单采血小板细菌的效果及血小板一系列生化和生理学指标的改变。方法将28ml浓度为500μmol／L核黄素加到250ml的单采血小板悬液中，再将一定浓度的G^＋菌（表皮葡萄球菌ATCC12228）和G^＋菌（大肠埃希菌ATCC25922）分别注入上述混悬液中，经265—370nm广谱紫外光6．2J／ml照射8～10min后，测定其浓度，并检测血小板体外参数的变化情况。结果经浓度50μmol／L的核黄素结合强度为6．2J／ml紫外光照射8—10min，可将一定浓度的模型菌灭活至〈1．22logCFU／ml。结论核黄素结合紫外光照射可以有效灭活细菌，而单采血小板的各项生化、生理学指标和阴性对照比较均差异无显著性。

关键词：核黄素/药理学光敏感药/药理学血小板输注

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Intracellular calcium dynamics during photolysis

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JOURNAL OF BIOMECHANICAL ENGINEERING-TRANSACTIONS OF THE ASME 1998年第5期120卷 570-578页

作者： Busch, NA Reiken, SR Toner, M Yarmush, ML Massachusetts Gen Hosp Ctr Engn Med Lab Surg Sci & Engn Boston MA 02114 USA Shriners Burns Hosp Boston MA 02114 USA Harvard Univ Sch Med Boston MA 02114 USA

The objective of this investigation was to gain a deeper understanding of the intracellular events that precede photolysis of cells. A model system, consisting of malignant melanoma cells pretreated,vith the calcium sensitive fluorescent dye, Fluo-3, was used to examine the intracellular calcium dynamics in single-cell photolysis experiments. Exposure of the cells to 632 nm laser light in the presence of photosensitizer, tin chlorin e6, resulted in a rise in intracellular calcium. The increase in intracellular calcium,was blocked using a variety of calcium channel blocking agents, including verapamil, nifedipine, and nickel. Treatment with the channel blockers was also effective in either decreasing or eliminating cell death despite the presence of lethal doses of photosensitizer and irradiation. These results show that intracellular calcium rises prior to plasma membrane lysis, and that this early rise in intracellular calcium is necessary for membrane rupture.

关键词：苯胺化合物/药代动力学钙/代谢钙通道阻滞药/药理学钙通道/药物作用钙通道/辐射效应钙信号/药物作用钙信号/辐射效应黑色素瘤金属卟啉类/药理学镍/药理学硝苯地平/药理学光化学疗法光解光敏感药/药理学光谱法质量基质辅助激光解吸电离时间因素肿瘤细胞培养的/辐射效应维拉帕米/药理学呫吨类/药代动力学人类

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Bactericidal activity of photocatalytic TiO₂ reaction:: Toward an understanding of its killing mechanism

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APPLIED AND ENVIRONMENTAL MICROBIOLOGY 1999年第9期65卷 4094-4098页

作者： Maness, PC Smolinski, S Blake, DM Huang, Z Wolfrum, EJ Jacoby, WA Natl Renewable Energy Lab Golden CO 80401 USA Univ Missouri Dept Chem Engn Columbia MO 65211 USA

When titanium dioxide (TiO2) is irradiated with near-UV light, this semiconductor exhibits strong bactericidal activity. In this paper, we present the first evidence that the lipid peroxidation reaction is the underlying mechanism of death of Escherichia call K-12 cells that are irradiated in the presence of the TiO2 photocatalyst, Using production of malondialdehyde (MDA) as an index to assess cell membrane damage by lipid peroxidation, we observed that there was an exponential increase in the, production of MDA, whose concentration reached 1.1 to 2.4 nmol - mg (dry weight) of cells(-1) after 30 min of illumination, and that the kinetics of this process paralleled cell death. Under these conditions, concomitant losses of 77 to 93% of the cell respiratory activity were also detected, as measured by both oxygen uptake and reduction of 2,3,5-triphenyltetrazolium chloride from succinate as the electron donor. The occurrence of lipid peroxidation and the simultaneous losses of both membrane-dependent respiratory activity and cell viability depended strictly on the presence of both light and TiO2. We concluded that TiO2 photocatalysis promoted peroxidation of the polyunsaturated phospholipid component of the lipid membrane initially and induced major disorder in the E. call cell membrane, Subsequently, essential functions that rely on intact cell membrane architecture, such as respiratory activity, were lost, and cell death was inevitable.

关键词：消毒/方法大肠杆菌/药物作用大肠杆菌/生长和发育光脂质过氧化作用/药物作用丙二醛/代谢氧耗量光敏感药/药理学光敏感药/辐射效应钛/药理学钛/辐射效应紫外线

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Bromohypericins are potent photoactive antiviral agents

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PHOTOCHEMISTRY AND PHOTOBIOLOGY 1999年第5期70卷 820-822页

作者： Hudson, JB Delaey, E de Witte, PA Univ British Columbia Dept Pathol & Lab Med Vancouver BC V5Z 1M9 Canada Katholieke Univ Leuven Lab Farmaceut Biol Fytofarmacol Louvain Belgium

Several hypericin derivatives, previously shown to have interesting light-mediated biological activities, were evaluated fur antiviral activities against herpes simplex virus and influenza virus. Three brominated hypericins, the dibromo- and tetrabromo-derivatives and the natural compound gymnochrome B were all very active against both viruses, particularly herpes simplex virus, although light was required in all cases for maximum activity. The dibromohypericin was the most potent, under standard assay conditions, gymnochrome B was approximately as active as hypericin if self and tetrabromohypericin significantly less so. Surprisingly, hexamethylhypericin, which is known to have potent anti-protein kinase (PK) C activity, as well as anticell proliferation properties, showed. no antiviral activity at all, The compounds were also evaluated in different serum concentrations, All the active compounds were inhibited by increasing concentrations of serum, but to different degrees, such that their relative antiviral potencies changed to some extent. Thus, in summary, there was no correlation between antiviral and anti-PK or anticellular activities, and consequently it is not possible at present to define those structural features of hypericin-type molecules that are required for their various biological activities.

关键词：抗病毒药/化学抗病毒药/药理学细胞系长尾猴黑疱疹病毒1型人/药物作用流感病毒A型/药物作用二萘嵌苯/类似物和衍生物二萘嵌苯/化学二萘嵌苯/药理学光敏感药/化学光敏感药/药理学构效关系 Vero细胞动物狗

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Oxidized low-density lipoprotein as a delivery system for photosensitizers: Implications for photodynamic therapy of atherosclerosis

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 1999年第1期289卷 528-534页

作者： de Vries, HE Moor, ACE Dubbelman, TMAR van Berkel, TC Kuiper, J Leiden Univ Sylvius Lab Leiden Amsterdam Ctr Drug Res Div Biopharmaceut NL-2300 RA Leiden Netherlands Leiden Univ Dept Mol Cell Biol NL-2300 RA Leiden Netherlands

Photodynamic therapy is a promising new strategy in the treatment of cardiovascular diseases. Photodynamic therapy for vascular diseases may be improved by the specific delivery of photosensitizers to the atherosclerotic lesion. In this study, we studied whether oxidatively modified low-density lipoprotein (OxLDL) could be used as a specific carrier for photosensitizers, thereby using the scavenger receptor expressed on macrophages as a target. The photosensitizer aluminum phthalocyanine chloride (AlPc) was incorporated into OxLDL, and its photodynamic effects were studied. Macrophages (RAW 264.7) were incubated with various concentrations of OxLDL-AlPc for different periods. After illumination of the cells with red right, cytotoxicity was observed that was dependent on the time of illumination and incubation. Macrophages incubated with OxLDL-AlPc that were not illuminated revealed no cytotoxicity;The uptake of the OxLDL-AlPc complexes was mediated by scavenger receptors expressed on macrophages. In the presence of the polyanion polyinosinic acid, a specific ligand for scavenger receptors,no cytotoxicity could be observed. Serum incubations of the OxLDL-AlPc complexes revealed that these complexes stay intact after incubation. No redistribution of AlPc to other plasma (lipo-) proteins could: be detected, and 80-90% of the AlPc remained associated with the OxLDL particle. These results indicate that OxLDL may function as a specific delivery system for photosensitizers to the scavenger receptors expressed on the macrophages in the atherosclerotic lesion, increasing the beneficial effects of photodynamic therapy for cardiovascular diseases.

关键词：动脉硬化/治疗细胞培养的色谱法液相药物释放系统乳状剂吲哚类/投药和剂量吲哚类/药理学脂蛋白类 LDL/代谢巨噬细胞/药物作用巨噬细胞/代谢有机金属化合物/投药和剂量有机金属化合物/药理学氧化还原光化学疗法光敏感药/投药和剂量光敏感药/药理学受体 LDL/药物作用受体 LDL/代谢受体氧化LDL 清道夫受体 E类甘油三酯类/化学甘油三酯类/分离和提纯动物小鼠

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Oxygen consumption through metabolism and photodynamic reactions in cells cultured on microbeads

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PHYSICS IN MEDICINE AND BIOLOGY 2000年第1期45卷 103-119页

作者： Schunck, T Poulet, P Univ Strasbourg 1 Inst Phys Biol Fac Med UPRESA 7004 CNRS F-67085 Strasbourg France

Oxygen consumption by cultured cells, through metabolism and photosensitization reactions, has been calculated theoretically. From this result, we have derived the partial oxygen pressure P-O2 in the perfusion medium Bowing across sensitized cultured cells during photodynamic experiments. The P-O2 variations in the perfusate during light irradiation are related to the rate of oxygen consumption through photoreactions, and to the number of cells killed per mole of oxygen consumed through metabolic processes. After irradiation, the reduced metabolic oxygen consumption yields information on the cell death rate, and on the photodynamic cell killing efficiency. The aim of this paper is to present an experimental set-up and the corresponding theoretical model that allows us to control the photodynamic efficiency for a given cell-sensitizer pair, under well defined and controlled conditions of irradiation and oxygen supply. To demonstrate the usefulness of the methodology described, CHO cells cultured on microbeads were sensitized with pheophorbide a and irradiated with different light fluence rates. The results obtained, i.e. oxygen consumption of about 0.1 mu M s(-1) m(-3) under a light fluence rate of 1 W m(-2), 10(5) cells killed per mole of oxygen consumed and a decay rate of about 1 h(-1) of living cells after irradiation, are in good agreement with the theoretical predictions and with previously published data.

关键词： CHO细胞细胞死亡/药物作用细胞死亡/辐射效应叶绿素/类似物和衍生物叶绿素/药理学仓鼠亚科光微球体模型理论氧耗量/药物作用氧耗量/辐射效应分压光化学疗法光敏感药/药理学动物

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Photonuclease activity of Taylor's blue

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BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 1999年第15期9卷 2249-2254页

作者： Mohammad, T Morrison, H Purdue Univ Dept Chem W Lafayette IN 47907 USA

Taylor's blue (1,9-dimethylmethylene blue, DMMB+) associates with DNA, at least in part, through intercalation as is evidenced from the red shift in the absorption maximum, diminution of the fluorescence, and induced circular dichroism in the presence of nucleic acid. Irradiation of DMMB+/covalently closed circular supercoiled Phi X174 phage DNA complex at lambda > 520 nm leads to DNA nicking in a dose-dependent manner. (C) 1999 Elsevier Science Ltd. All rights reserved.

关键词：圆二色性 DNA/药物作用 DNA/代谢 DNA/辐射效应 DNA损伤嵌入剂/代谢嵌入剂/药理学亚甲蓝/类似物和衍生物亚甲蓝/代谢亚甲蓝/药理学光敏感药/代谢光敏感药/药理学光谱分析

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Effect of mammalian cell differentiation on response to exogenous 5-aminolevulinic acid

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PHOTOCHEMISTRY AND PHOTOBIOLOGY 1999年第2期69卷 231-235页

作者： Li, G Szewczuk, MR Pottier, RH Kennedy, JC Queens Univ Dept Oncol Kingston ON K7L 3N6 Canada Queens Univ Dept Microbiol & Immunol Kingston ON K7L 3N6 Canada Royal Mil Coll Canada Dept Chem & Chem Engn Kingston ON Canada

Many different types of mammalian cells accumulate fluorescing and photosensitizing concentrations of protoporphyrin IX (PpIX) when exposed to exogenous 5-aminolevulinic acid (ALA) in vivo or in vitro. Most types of malignant cells accumulate substantially more ALA-induced PpIX than do the normal cells from which they arose. Most types of malignant cells also are less differentiated than their normal counterparts. We therefore considered the possibility that malignant cells demonstrate a malignant ALA phenotype (accumulate abnormally large amounts of PpIX when exposed to exogenous ALA) as a direct consequence of their less differentiated state, Human promyelocyte cell lille HL-60 and mouse preadipocyte cell line 3T3 L1 were induced to differentiate by exposing them to inducing agents in vitro, The HL-60 cells accumulated less ALA-induced PpIX when differentiated, but the 3T3 L1 cells accumulated more. It appears then that changes in the ALA phenotype with changes in the state of differentiation are cell type specific. The decreased accumulation of ALA-induced PpIX that accompanied differentiation of the promyelocytic leukemia cells may have clinical application for rapid quantitation of the response of myelocytic leukemia patients to differentiation therapy.

关键词： 3T3细胞/药物作用氨基酮戊酸/药理学细胞分化细胞分离流式细胞术 HL-60细胞/药物作用表型光敏感药/药理学动物人类小鼠

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Cyanine dyes as protectors of K562 cells from photosensitized cell damage

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PHOTOCHEMISTRY AND PHOTOBIOLOGY 2000年第4期71卷 413-421页

作者： Sima, PD Kanofsky, JR US Dept Vet Affairs Vet Affairs Edward Hines Jr Hosp Res Serv Hines IL 60141 USA US Dept Vet Affairs Vet Affairs Edward Hines Jr Hosp Med Serv Hines IL 60141 USA Loyola Univ Stritch Sch Med Dept Med Maywood IL 60153 USA Loyola Univ Stritch Sch Med Dept Mol & Cellular Biochem Maywood IL 60153 USA

Several cyanine dyes were found to protect K562 leukemia cells against toxicity mediated by cis -di(4-sulfonatophenyl)diphenylporphine (TPPS2) and light. Most cyanine dyes derived from dimethylindole were better photoprotectors than cyanine dyes with other structures. This correlated with the fact that cyanine dyes derived from dimethylindole were predominately monomeric at millimolar concentrations within K562 cells, while other cyanine dyes formed aggregates. For cyanine dyes that are derived from dimethylindole and have absorption band wavelengths greater than 700 nm, fluorescence-energy transfer from TPPS2 to the cyanine dye was the most important mechanism for photoprotection, There was no spectroscopic evidence for complex formation between the cyanine dyes and TPPS2, The dimethylindole derivative, 1,1',3,3,3',3'-hexamethylindodicarbocyanine, was an excellent photoprotector, but a poor quencher of TPPS2 fluorescence and a relatively poor singlet-oxygen quencher. This cyanine dye may act by quenching excited triplet TPPS2. Singlet-oxygen quenching may contribute to the photoprotection provided by cyanine dyes not derived from dimethylindole. Differences in the subcellular distribution of the various cyanine dyes studied may have contributed to the different apparent mechanisms of photoprotection.

关键词：细胞存活/药物作用着色剂/化学着色剂/药理学氰化物/化学 K562细胞光光敏感药/药理学卟啉类/药理学辐射防护剂/化学辐射防护剂/药理学人类

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Light-induced apoptosis involves a defined sequence of cytoplasmic and nuclear calcium release in AlPcS₄-photosensitized rat bladder RR 1022 epithelial cells

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PHOTOCHEMISTRY AND PHOTOBIOLOGY 2000年第2期72卷 210-216页

作者： Rück, A Heckelsmiller, K Kaufmann, R Grossman, N Haseroth, E Akgün, N Inst Laser Technol Med & Metrol D-89081 Ulm Germany Univ Frankfurt Clin Dermatol & Venereol D-6000 Frankfurt Germany Ben Gurion Univ Negev Fac Hlth Sci IL-84105 Beer Sheva Israel Soroka Univ Med Ctr Skin Bank & Investigat Dermatol Lab Beer Sheva Israel

Oxidative stress induced by light activation of photosensitizers is regarded to have a role in triggering cell death pathways during photodynamic therapy (PDT). Reactive oxygen species have been proposed to act as signal transduction molecules activating downstream reactions that lead to apoptosis, Mainly debated is the cooperating role of other signaling systems like calcium or pH. The present work contributes to this discussion by studying PDT effects in cell cultures of rat bladder epithelial cells for the hydrophilic tetrasulfonated aluminum phthalocyanine (AlPcS4), Cells were coincubated with the photosensitizer and the calcium-sensitive probe Fluo-3, The light-induced reactions were analyzed with a confocal laser scanning microscope. The dynamics of the process during light activation was observed with subcellular resolution. A transient calcium elevation during the irradiation process was detected, especially in the cell's nuclei, followed by a more sustained increase. The evaluation of the energy-dose-dependent phototoxicity after an incubation time with the photosensitizer of I and 24 h, showed enhanced phototoxicity when the drug was present for 24 h, Surprisingly, stimulation of cell proliferation was observed at very low light doses (at 0.2 J/cm(2)) when the drug was incubated for 24 h (cell viability 160%), Induction of apoptosis could be observed after irradiation with fluences between 1 and 3 J/cm(2). Apoptotic cells were identified with fluorescein isothiocyanate-labeled Annexin V, which binds to phosphatidylserine after its translocation to the outer plasma membrane. In the presence of the antioxidant pyrrolidinedithiocarbamate the transient calcium elevation was totally inhibited, as was the subsequent translocation of PS, In contrast, N-acetyl-L-cysteine did not suppress the transient calcium increase. Our data might be consistent with calcium regulated processes during AlFcS(4)-PDT and the involvement of oxygen radicals.

关键词：细胞凋亡/生理学细胞凋亡/辐射效应钙信号/辐射效应细胞系细胞核/代谢细胞质/代谢上皮细胞/药物作用上皮细胞/代谢上皮细胞/辐射效应吲哚类/药理学光有机金属化合物/药理学光化学疗法光敏感药/药理学膀胱/药物作用膀胱/代谢膀胱/辐射效应动物大鼠

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