Caracterização dos efeitos tóxicos do 1, 2-dihidroxibenzeno em células originadas do sistema nervoso: investigação do efeito protetor de derivados de plantas / Characterization of toxic effects of 1, 2-dihydroxybenzene cells originating in the nervous system: investigating the protective effect of plant-derived

Catecóis são derivados do benzeno, podendo apresentar citotoxicidade, que pode constituir um modelo experimental útil para o desenvolvimento de novos fármacos. No bioma brasileiro inúmeras plantas produzem metabólitos com atividades diversas, como antioxidantes, ou inibidores do crescimento celular. No Brasil, as neoplasias são a segunda causa de óbito, especialmente aquelas derivadas do sistema nervoso, aumentando o interesse por novos antineoplásicos e agentes neuroprotetores. Este trabalho caracteriza efeitos citotóxicos do 1,2-dihidroxibenzeno (CAT) e discretamina (DSC) em células do sistema nervoso in vitro. Determinou-se a EC50 de CAT e DSC usando brometo de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolium (MTT), investigou-se sua auto-oxidação por espectrofotometria, avaliou-se mudanças morfológicas e condensação/fragmentação nuclear por microscopia. Avaliou-se a proteção de DSC e 8-metoxipsoraleno (8-MOP) contra a citotoxicidade do CAT. O padrão de morte celular foi analisado por citometria de fluxo. A espoliação de glutation reduzido (GSH) foi analisada usando monoclorobimano. A toxicidade do CAT para células SH-SY5Y e C6 depende da dose e associa-se à formação de quinonas. Houve mudanças morfológicas, condensação/fragmentação da cromatina e morte apoptótica, não relacionada à espoliação de GSH. DSC não foi tóxica para células SH-SY5Y, porém protegeu contra os efeitos do CAT em baixas concentrações. DSC mostrou-se citotóxica para células de glioma (GL-15 e C6) e potencializou o CAT. Pré-tratamento por 30 minutos com DSC protegeu contra a ação do CAT após 72 horas. 8-MOP potencializou os efeitos do CAT, não revertendo seus efeitos na viabilidade celular, morfologia celular, condensação/fragmentação nuclear, e espoliação de GSH. Esses resultados caracterizam um modelo de citotoxicidade que pode ser aplicado no desenvolvimento de novos agentes farmacológicos. Estudos complementares são necessários para elucidar a proteção da DSC.

Catechols are benzene derivatives, which may exhibit cytotoxic activity that can be employed to develop new drugs. Plants are important sources of metabolites with pharmacological activities such as antioxidants, or cell growth inhibitors. In Brazil, cancer is the second leading cause of death, especially those derived from the nervous system, which increase the interest for new antineoplastic and neuroprotective drugs. The cytotoxic effects promoted by 1,2-dihydroxybenzene (CAT) and discretamine (DSC) in nervous system cells were characterized in vitro. The protective effects of DSC and 8-methoxypsoralen (8-MOP) against CAT-induced cytotoxicity were also evaluated. CAT and DSC EC50 was determined by using 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT). CAT auto-oxidation was investigated by spectrophotometry. Morphological changes and nuclear condensation/ fragmentation were evaluated by microscopy. The pattern of cell death was obtained by flow cytometry. Reduced glutathione (GSH) depletion was analyzed by using monochlorobimane. CAT induced a dose-dependent toxicity to SH-SY5Y and C6 cells, associated with reactive quinones formation. It also induced morphological changes, nuclear condensation/fragmentation, and apoptotic death not caused by GSH depletion. DSC was not toxic to SH-SY5Y cells, but protected against CAT effects at low concentrations. DSC was be cytotoxic to glioma cells (GL-15 and C6) and potentiated CAT effects. However, pretreatment for 30 minutes with DSC protected them against CAT after 72 hours. 8-MOP also potentiated CAT effects instead to protect cells. These results characterize an experimental model useful for studies searching new pharmacological agents. However, further studies are needed to elucidate the DSC protective effects.

Establishment of mus skin photo-damage model by 8-MOP plus UVA irradiation / 华中科技大学学报（医学）（英德文版）

To establish a simple and reliable animal model of skin photo-damage, 20 mice were treated with 8-MOP and exposed to UVA (UVA 320-400 nm) for 24 h. After irradiation, the structure of the epidermis and dermis, collagen fibers, elastic fibers were observed by using HE staining and Weigert technique and compared with the normal controls. The acanthosis and epidermis proliferation with accompanying hyperkeratosis and parakeratosis were observed. Inflammatory infiltration was noted in the dermis. The elastic fibers became coarse, irregularly arranged and clustered, with their number increased. The collagen fibers showed obvious degeneration and some amorphous materials could also be observed. The blood vessels were irregularly dilated and vascular walls were thickened, with infiltration of inflammatory cells. It is concluded that murine photodamage model can be quickly, conveniently and reliably established by means of 8-MOP/UVA.

study of pentobarbitone-methoxsalen interaction in rats

Experiments carried out to study pentobarbitonemethoxsalen interaction in rats. The study was conducted on adult albino rats [150-250 g, either sex] in two phases. In phase I, methoxsalen was administered [5, 10 and 15 mg/kg i.p. x 5 days] and pentobarbitone was administered [30 mg/kg i.p.] 24 hours after the last dose of methoxsalen. In phase II, methoxsalen was administered [10 mg/kg p.o. x two weeks] and pentobarbitone was administered [in dose of 15, 20 and 30 mg/kg i.p.] 24 hrs after the last dose of methoxsalen, CNS activity such as pinna, sound and righting reflex were observed. In phase I, pentobarbitone caused CNS depression in methoxsalen treated and control groups. In phase II, pentobarbitone in dose of 15, 20 and 30 mg/kg in methoxsalen treated rats also caused CNS depression in dose dependent manner. The action of pentobarbitone was potentiated in methoxsalen treated rats hence barbiturates whenever indicated in patients on methoxsalen, should be used with caution

2-Deoxy-D-glucose induced modulation of DNA damage repair, survival, mutagenesis and recombinogenesis in 8-MOP+UVA treated yeast.

Cellular and genomic effects of post-treatment repair modulation by 2-deoxy-D-glucose (2-DG) and yeast extract were studied in 8-MOP + UVA treated cells of Saccharomyces cerevisiae. The type of lesions and their repair in phosphate buffer glucose (PBG) differed with UVA dose. At low UVA dose (1.4 kJ/m2), lesions were sublethal and mutagenic and did not repair by recombinogensis. The fraction of potentially lethal lesions and lesions repaired by recombinogenesis increased with UVA dose. Cellular repair in PBG was largely error-free and was inhibited by 2-DG. Yeast extract enhanced cellular repair and also recombinogensis; 2-DG in presence of yeast extract promoted error-prone repair. Pulsed-field gel electrophoresed chromosomal DNA bands did not show observable alterations immediately after 8-MOP + UVA treatment. On post-treatment incubation in PBG, the intensity ratio (rho n), of each band altered in a biphasic manner showing decrease first, followed by either increase or no change upto 24 hr depending upon UVA exposure dose. Presence of 2-DG in PBG inhibited decrease in rho n in a concentration dependent manner. Yeast extract reduced the time of first phase of DNA repair. 2-DG and yeast extract together reduced the time of first phase of repair and also inhibited the subsequent increase in rho n, which was observed in the case of yeast extract in PBG. It is proposed that (i) 2-DG in PBG inhibits excision of DNA damage and error-free repair; (ii) yeast extract stimulates the error-prone repair associated with cell cycle and recombinogenesis; (iii) 2-DG in presence of yeast extract allows excision of damage but inhibits build up through recombinogenesis inducing instead, cell cycle associated error-prone repair. A simple schematic model has been proposed to explain these events.

Laboratory and clinical evaluation of a new formulation of 8-methoxypsoralen in the treatment of vitiligo

A new formulation of 8-methoxypsoralen [8-MOP] capsule was investigated both in the laboratory and clinically. Laboratory investigations showed that the new formulation is absorbed better than the currently available 8-MOP and it reaches a peak blood level in 45 minutes. Clinically, 14 vitiligo patients received the new drug in a dose ranging from 0.25-0.4 mg/kg body weight and exposed their vitiliginous patches to the sun at midday [45 minutes after drug ingestion]. An excellent response [>75% of a test patch repigmented] was obtained in 5 patients [35.7%], a good response [50-75% of the patch covered] was obtained in 3 patients [21.4%] and an unsatisfactory response [<50% of the patch covered] was obtained in 6 patients [42.9%]. Side effects were very minimal. The new formulation of 8-MOP capsule is safe, more effects than the marketed tablet formulation and its dose can be raised in nonresponding cases without producing nausea or vomiting

Quantitative and Morphologic Changes of Epidermal Langerhans Cell after Photochemotherapy in Guinea Pigs

We studied the number and morphologic alterations of ATPase-positive Langerhans cells (LCs) in guinea pig epidermal sheets and also performed electron microscopic observations on them following long-term systemic psoralen photochemotherapy (PUVA) using 8-methoxypsoralen (8-MOP) and trimethylpsoralen (TMP). We con-firmed that the LCs are sensitive to PUVA treatment. The LC depleting effects of PUVA are dose-related and the restorator1 of LCs takes place by the 4th week following cessation of PUVA. Neither psoralens alone nor UVA alone showed any effects on LCs. There were no significant differences between 8-MOP and TMP when comparing the effects of 8-MOP plus UVA and TMP plus UVA on LC numbers and morphology. It seems that a moderate dose of PUVA causes an actual decrease in the number due to cell damage. Also, it appears likely that giant LCs appear during and following PUVA treatment as a compensatory process of the remaining cells.