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21.
Poly(ADP-ribosyl)ation of nuclear proteins is catalyzed by poly(ADP-ribose) polymerase. This enzyme is involved in the regulation of basic cellular functions of DNA metabolism. DNA breaks induced by DNA-damaging agents trigger the activation of poly(ADP-ribose) polymerase increasing its endogenous level. This increase modifies the pattern of poly(ADP-ribosyl)ated chromatin proteins. In this paper we describe a procedure for the isolation of intact nuclei from rat liver to be used for the endogenous activity assay. Artifactual activation of the enzyme was avoided since a very low level of DNA-strand breaks occurs during the isolation of nuclei. We present a series of experiments which prove the ability of this procedure to detect increases in endogenous liver activity without modification of the total level. The application of this technique can be useful for a better understanding of the role of early changes in poly(ADP-ribose) polymerase level in physiological conditions and during exposure to DNA-damaging agents. 相似文献
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C F Cesarone L Scarabelli P Giannoni G Gallo M Orunesu 《Biochemical and biophysical research communications》1990,171(3):1037-1043
Previous studies have demonstrated that an increase in poly(ADP-ribose) polymerase activity could be closely related to DNA replication during liver regeneration and to DNA repair synthesis in different experimental systems. This relationship was further investigated by studying the time course of endogenous and total poly(ADP-ribose) polymerase activity in cultured rat hepatocytes stimulated by epidermal growth factor. This mitogen has been shown to stimulate DNA synthesis in liver cells both in vivo and in vitro. A 6-fold increase in endogenous activity was observed early after epidermal growth factor addition, just before DNA synthesis. A subsequent 4-fold increment in total enzyme activity, concomitant with DNA synthesis, was detected. Orotic acid, which has recently shown mitoinhibitory effect, abolished the epidermal-growth-factor-induced increase in endogenous and total poly(ADP-ribose) polymerase activity, as well as DNA synthesis. On the contrary, 3-aminobenzamide inhibitor of poly(ADP-ribose) polymerase completely suppressed the endogenous activity but only partially modified the increase in total catalytic level and the overall pattern of thymidine incorporation. Taken together, these data indicate that, in cultured hepatocytes, the induction of DNA synthesis is supported by an increased poly(ADP-ribose) polymerase activity. 相似文献
23.
Degan P Sancandi M Zunino A Ottaggio L Viaggi S Cesarone F Pippia P Galleri G Abbondandolo A 《Journal of cellular biochemistry》2005,94(3):460-469
Exposure of freshly drawn lymphocytes and lymphoblastoid cells (LB and COR3) to simulated microgravity decreased the intracellular ATP concentration to 50%-40% of the value found in normal growth conditions. The decrease was reversible although recovery to normal values occurred only slowly both in lymphocytes and in lymphoblastoid cells. Poly(ADP-ribose) polymerase (PARP ) activity was increased indicating that cells exposed to conditions of reduced gravitation experience stress. Exposure to microgravity forces cells into a condition of metabolic quiescence in which they appear to be particularly sensitive to subsequent exposures to a genotoxic agent. Thus, treatment of cells with the strong redox agent potassium bromate under microgravity conditions, indicated an impairment in repair of DNA 8-hydroxy-2'-deoxyguanosine (8-OHdG), an oxidized derivative of deoxyguanosine. We conclude that gravitational modulation of the kind routinely obtained under laboratory conditions and during spaceflights is a stressful process to which cells appear to be extremely sensitive. These effects may reflect the physiological alterations observed in astronauts and in animals following spaceflights or exposure to conditions of simulated microgravity. 相似文献
24.
Cesarone CF Camboni MG Meloni MA Cogoli A Pippia P 《Journal of gravitational physiology : a journal of the International Society for Gravitational Physiology》2001,8(1):P23-P24
Human peripheral blood lymphocytes (PBL), activated with concanavalin A (ConA), were used to determine the effects of simulated microgravity on poly(ADP-ribose) polymerase (PARP) activity. Results indicate that the ConA stimulation of human cultured PBL induces a partial but signitficant inhibition of PARP-1 acitvity (-30%). In control PBL, not exposed to ConA, after 24 hours, there was a clear decrease in PARP-1 acitivty (-40%). In PBL exposed to ConA and simulated weightlessness, activity decreased by -37%. 相似文献
25.
F De Martino A De Matteo V L'Abbate A Soricelli A Fontana E Petrenga M Cesarone 《Bollettino della Società italiana di biologia sperimentale》1980,56(8):809-815
The authors have tested serum levels of PRL after administration of atropine 10' and 60' before stimulation test with TRH, in 12 male and 12 female. The administration of the drug 10' before TRH has inhibited the PRL release, in all the groups. No effect presented atropine if administered 60' before TRH test. 相似文献
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27.
M C Perrone D Serra F D'Agostini C F Cesarone 《Bollettino della Società italiana di biologia sperimentale》1991,67(9):875-879
Male wistar rats were treated with a diet supplemented with 0.05% 2-acetylaminofluorene (2AAF) and/or 0.2% N-acetylcysteine (NAC) according to the protocol of Teebor and Becker. Eleven haematochemical parameters were evaluated at the third week of the first two cycles. The results showed a slight yet significant decrease in total proteins and triglycerides, and an increase in total bilirubin, gamma-glutamyltranspeptidase and alkaline phosphatase, as compared to untreated controls. Co-treatment with NAC slightly attenuated the alterations induced by 2AAF. On the whole, these results demonstrate that 2AAF is poorly necrotic to hepatocytes, and hence its known ability to damage the liver appears to mainly depend on nuclear effects rather than on cytoplasmic changes. 相似文献