共查询到20条相似文献,搜索用时 31 毫秒
1.
Formation and subsequent removal of O6-methylguanine from deoxyribonucleic acid in rat liver and kidney after small doses of dimethylnitrosamine 总被引:5,自引:0,他引:5
下载免费PDF全文
![点击此处可从《The Biochemical journal》网站下载免费的PDF全文](/ch/ext_images/free.gif)
1. The amounts of 7-methylguanine and O6-methylguanine present in the DNA of liver and kidney of rats 4h and 24h after administration of low doses of dimethylnitrosamine were measured. 2. O6-Methylguanine was rapidly removed from liver DNA so that less than 15% of the expected amount (on the basis of 7-methylguanine found) was present within 4h after doses of 0.25mg/kg body wt. or less. Within 24h of administration of dimethylnitrosamine at doses of 1mg/kg or below, more than 85% of the expected amount of O6-methylguanine was removed. Removal was most efficient (defined in terms of the percentage of the O6-methylguanine formed that was subsequently lost within 24h) after doses of 0.25–0.5mg/kg body wt. At doses greater or less than this the removal was less efficient, even though the absolute amount of O6-methylguanine lost during 24h increased with the dose of dimethylnitrosamine over the entire range of doses from 0.001 to 20mg/kg body wt. 3. Alkylation of kidney DNA after intraperitoneal injections of 1–50μg of dimethylnitrosamine/kg body wt. occurred at about one-tenth the extent of alkylation of liver DNA. Removal of O6-methylguanine from the DNA also took place in the kidney, but was slower than in the liver. 4. After oral administration of these doses of dimethylnitrosamine, the alkylation of kidney DNA was much less than after intraperitoneal administration and represented only 1–2% of that found in the liver. 5. Alkylation of liver and kidney DNA was readily detectable when measured 24h after the final injection in rats that received daily injections of 1μg of [3H]dimethylnitrosamine/kg for 2 or 3 weeks. After 3 weeks, O6-methylguanine contents in the liver DNA were about 1% of the 7-methylguanine contents. The amount of 7-methylguanine in the liver DNA was 10 times that in the kidney DNA, but liver O6-methylguanine contents were only twice those in the kidney. 6. Extracts able to catalyse the removal of O6-methylguanine from alkylated DNA in vitro were isolated from liver and kidney. These extracts did not lead to the loss of 7-methylguanine from DNA. 7. The possible relevance of the formation and removal of O6-methylguanine in DNA to the risk of tumour induction by exposure to low concentrations of dimethylnitrosamine is discussed. 相似文献
2.
Effects of ecdysone on a Drosophila cell line 总被引:3,自引:0,他引:3
R Rosset 《Experimental cell research》1978,111(1):31-36
When treated by ecdysone, Drosophila cells of the line Kc start aggregating in clumps 24 h later. Different early events occur before this morphological process: DNA synthesis and mitosis stop after 8–10 h; RNA synthesis after a transient stimulation is progressively inhibited. 相似文献
3.
Cell suspensions were prepared from normal and regenerating liver of adult rats by perfusion with a calcium-chelating agent (EGTA), collagenase and hyaluronidase, and the cells were incubated in culture medium. In cultures prepared from regenerating liver at 20 h after partial hepatectomy, 23 ± 4% of parenchymal cells initially incorporated [3H]TdR. This incorporation was shown to reflect semiconservative DNA replication. At least some parenchymal cells were able to complete their DNA synthesis and to progress through G2 and mitosis. Numbers of hepatocytes in mitosis increased up to 12 h of culture. On the other hand, no entry of hepatocytes into the S period was detectable in cultures prepared from normal or regenerating liver. 相似文献
4.
G P Margison 《Chemico-biological interactions》1982,38(2):189-201
The effect of pretreatment of rats with various symmetrical dialkylnitrosamines on the repair of O6-methylguanine produced in liver DNA by a low dose of [14C]dimethylnitrosamine (DMN) has been examined. DMN, diethylnitrosamine (DEN), dipropylnitrosamine (DPN) or dibutylnitrosamine (DBN) were administered to rats for 14 consecutive weekdays at a daily dose of 5% of the LD50. Animals were given [14C]DMN 24 h after the last dose and were killed 6 h later. DNA was extracted from the liver and analysed for methylpurine content after mild acid hydrolysis and Sephadex G-10 chromatography. While the amounts of 3-methyladenine and 7-methylguanine were only slightly different from controls, the amounts of O6-methylguanine in the DNA of the dialkylnitrosamine pretreated rats were about 30% of those in control rats, indicating a considerable increase in the capacity to repair this base. Liver ribosomal RNA from control and dialkylnitrosamine pretreated rats contained closely similar amounts of O6-methylguanine suggesting that the induced enzyme system does not act on this base in ribosomal RNA in vivo. Pretreatment with these dialkylnitrosamines also enhanced the repair of O6-methylguanine in liver DNA when they were given as a single dose (50% of the LD50) either 3 or 7 days before the [14C]DMN. In addition, single low doses of DMN or DEN (5% of the LD50) given either 1 or 6 days before [14C]DMN increased O6-methylguanine repair and the magnitude of the effect after DEN was similar to that produced by the other pretreatment schedules. The possible mechanism(s) of the induction of O6-methylguanine repair and its relation to hepatotoxicity, DNA alkylation, carcinogenesis and the adaptive response in Escherichia coli are discussed. 相似文献
5.
We investigated whether acute iron intoxication causes oxidative DNA damage, measured in terms of 7-hydro-8-oxo-2′-deoxyguanosine, 8-oxodG, in nuclear DNA in testes and epididymal sperm cells in vivo and in vitro in rats. In addition, we investigated levels of the modified nucleoside in liver and kidney and measured its urinary excretion.Sperm cells were isolated from the epididymides and the testes cells were isolated after homogenisation. In vitro, the sperm and testes cells were incubated with increasing concentrations of FeCl2 ranging from 0 to 600 μM. The median (range) levels of 8-oxodG/105 dG in the epididymal sperm cells increased from 0.48 (0.42–0.90) to 15.1 (11.4–17.6) (p < 0.05), whereas the level rose from 0.63 (0.22–0.81) to 8.8 (4.5–11.6) (p < 0.05) at 0 and 600 μM, respectively, in the testicular cells.In vivo groups of 7–8 rats received 0, 200 or 400 mg iron/kg as dextran i.p. After 24h, epididymal sperm cells, testes, kidneys and liver were collected for analysis. Kidney and sperm DNA showed a significant increase in 8-oxodG in the iron-treated animals. The median (range) values of the 8-oxodG/105 dG in the epididymal sperm cells rose from 0.66 (0.38–1.09) to 1.12 (0.84–5.88) (p < 0.05) at 0 and 400 mg iron/kg, respectively, whereas the values in the testes and liver showed no significant change. In the kidneys the 8-oxodG/105 dG median (range) values were 0.98 (0.73–1.24), 1.21 (1.13–1.69) and 1.34 (1.12–1.66) after 0, 200 and 400 mg iron/kg, respectively (p < 0.05).The 8-oxodG-excretion rate was measured in 24 h urine before and after iron treatment. The rate of urinary 8-oxodG excretion increased from 129 (104–179) pmol/24 h before treatment to 147 (110–239) pmol/24h after treatment in the group receiving 400 mg iron/kg (p < 0.05).The results indicate that acute iron intoxication may increase oxidative damage to sperm and kidney DNA. 相似文献
6.
A number of growth phenomena observed in vitro have shown that cells, at high densities, produce and release substances which, when they have reached a given concentration, arrest further growth. In vivo, these possibilities can be studied on the model of rapid regeneration of the rat liver after 65-70% partial hepatectomy (PH). We evaluated the course of liver regeneration after PH in animals treated with dialysates (DIA) of intact rat tissues. In addition to kidney and lymph node DIA, we were particularly interested in the effect of liver and spleen DIA. The experiments were carried out on male rats weighing 210-240 g. The relevant DIA was administered 24 h prior to PH; the controls were given physiological saline. The animals were killed just before PH and 24, 48, 30 and 72 h and 14 days after. DIA obtained from intact liver tissue inhibited the regeneration process induced by PH and its effect persisted 48 h after PH. Compared with the controls and with the rats given kidney DIA, DNA synthesis in the liver 24 h after PH was reduced to 77%. After spleen DIA, several (still hypothetical) factors probably acted together synergically (factors belonging to the immune system--RES--and spleen-produced factors capable of promoting proliferation of the hepatocytes--the "portal blood factor"). We arrived at this conclusion from an evaluation of liver DNA synthesis 24 h after 24h after PH, when synthesis was altogether markedly raised, but attained far higher values after the administration of spleen DIA.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
7.
Marios Marselos Riitta Törrönen Timo Koivula Martti Koivusalo 《Biochimica et Biophysica Acta (BBA)/General Subjects》1979,583(1):110-118
There is a genetically determined variation in the inducibility of a high-Km cytoplasmic aldehyde dehydrogenase activity in the rat liver by treatment with phenobarbital. In the present experiments this activity increased after phenobarbital administration in the phenobarbital-responsive rats also in the intestinal postmitochondrial supernatant fraction. Phenobarbital-nonresponsive rats did not exhibit such an increase after drug treatment. Intraperitoneal administration of 2,3,7,8,-tetrachlorodibenzo-pdioxin, strongly enchanced the cytoplasmic enzyme activity in the liver of both responsive and nonresponsive rats. This effect was also seen in the serum but not in the intestinal or hte kidney. Intragastric administration of 3-methylcholanthrene, 3,4,-benzpyrene or chrysene induced the activity in liver and intestine but not in serum or kidney. The activity in liver was also induced by long-term feeding with 2-acetamido-fluorene. The activities induced by tetrachlorodibenzodioxin or the carcinogens had similar behaviour in isoelectric focusing in gel slabs and in gel chromatography, suggesting a possible common identity of these induced enzymes. The activity induced by these agents could be clearly differentiated both from the activity induced by phenobarbital and from the normal cytoplasmic activities. 相似文献
8.
A E Pegg 《Chemico-biological interactions》1978,22(1):109-116
Pretreatment with diethylnitrosamine or dipropylnitrosamine increased the amount of labelled O6-methylguanine found in liver DNA 4 and 24 h after injection of 10 μg/kg [3H] dimethylnitrosamine. Dibutylnitrosamine treatment had a similar, though smaller effect at 4 h but was ineffective when measurements were made 24 h after the dimethylnitrosamine was given. These pretreatments did not affect 7-methylguanine levels in the DNA showing that the metabolic conversion of dimethylnitrosamine into a methylating agent was not altered. Previous studies have shown that O6-methylguanine is rapidly removed from hepatic DNA after methylation to a small extent but removal is less efficient after higher amounts of methylation. Therefore, the most probable explanation for the present findings is that these longer dialkylnitrosamines produce a similar product in DNA which interferes with the loss of O6-methylguanine. This hypothesis was supported by experiments showing that diethylnitrosamine did give rise to O6-ethylguanine which was lost from the DNA at a rate comparable to the observed loss of O6-methylguanine in diethylnitrosamine pretreated rats. This method may, therefore, be of value for determination of whether other nitrosamines, not available in a radioactively labelled form, react with DNA at external oxygen atoms. The present results also suggest that different dialkylnitrosamines might have additive effects in prolonging damage to DNA which could be important in carcinogenesis. 相似文献
9.
Effects of a single intraperitoneal injection of cathepsin D (CatD) on DNA synthesis and mitosis in the mouse liver and kidney were investigated. Twenty micrograms of catD induced a significant stimulation of DNA synthesis in the liver, but not in the kidney, in a dose-dependent fashion and with a peak activity at 38 h after the injection. CatD also stimulated liver mitosis, with a peak value at 44 h after the injection. 相似文献
10.
Cultures of epithelial rat kidney cells were used to investigate the effect of lead on protein and DNA metabolism. Lead adapted cells were cultured in 5 and 10 μM Pb(NO3)2 for several months and subsequently showed the same growth characteristics but a higher resistance to increased lead doses than untreated cells. Both normal and adapted cell types were exposed to lead nitrate in concentrations from 5 to 1000 μM for incubation periods of from 1 to 24 h. 3H-leucine incorporation was increased in 10 μM lead by up to 140% in normal but not in adapted cells, pointing to a resistance forming mechanism. Not only is the overall rate of protein synthesis increased, but separations of cytosolic proteins by SDS-polyacrylamide electrophoresis indicate the novel or increased synthesis of several proteins. A difference could also be observed in the DNA metabolism, where normal cells show a very pronounced increase of up to 900% after 24 h of incubation. This was not observed in adapted cells. 相似文献
11.
The binding of ethyl carbamate labelled with carbon-14 in the alkyl or carbonyl group, and of methyl, n-butyl and n-propyl carbamates labelled in the alkyl group, to the DNA of mouse liver, lung and kidney has been studied in male Crackenbush mice. Only ethyl carbamate bound to liver and kidney DNA to any significant extent.The binding of ethyl carbamate labelled with carbon-14 in the C1, C2 or the carbonyl position was examined and compared. The levels of binding of [1-14C]- and [2-14C]ethyl carbamate to liver DNA were not significantly different (328 ± 34 and 267 ± 24 dpm/mg DNA, respectively), but there was very little binding of the [carbonyl-14C]ethyl carbamate (26 ± 3 dpm/mg DNA). Furthermore, only 18% of the radioactivity was removed from the DNA labelled with the alkyl-labelled carbamates, whereas 65% of the radioactivity was removed from the DNA labelled with carbonyl-labelled ethyl carbamate on continuous ether extraction. It was concluded that the bound molecule does not contain the carbonyl carbon and is probably an ethyl group. 相似文献
12.
Summary Incorporation of thymidine into nuclear and mitochondrial DNA has been measured in the livers of normal rats and of rats killed 17 h or 24 h after partial hepatectomy. When total-body X-irradiation (500 or 1,500 R) is given at increasing intervals (up to 30 h) before sacrifice, a progressive decrease followed by a plateau of low level of incorporation is observed in the nuclear DNA (N-DNA) whereas the synthesis of mitochondrial DNA (M-DNA) first decreases until a minimal value is reached after 4 h in normal liver and 12 h in regenerating liver, then recovers at an exponential rate, quite independently of the nuclear DNA synthesis.The results suggest that irradiation can interfere with the enzymatic processes of both M-DNA and N-DNA syntheses, but that the persistence of the inhibition of N-DNA synthesis must be ascribed to a temporary block in the cell cycle.The SH-protector AET injected to non-irradiated rats exerts a very small inhibitory effect on normal liver N-DNA and M-DNA synthesis. In regenerating liver, both syntheses are affected rather similarly suggesting a common mechanism of inhibition through impairment of some step in the synthetic process. 相似文献
13.
A large dose of dimethylnitrosamine was administered to rats by two different dosing regimens, one being eleven intraperitoneal injections of 5 mg/kg body wt. at 12-h intervals (a dosing regimen strongly carcinogenic for the kidney but not the liver), and the other being a continuous dosing over several weeks by adding 8.5 mg of dimethylnitrosamine to each litre of drinking water giving an approximate daily dose of 0.7 mg/kg body wt. This treatment is known to be strongly carcinogenic for the liver but not the kidney. The accumulation in DNA of liver and kidney of the methylated purines 7-methylguanine and O6-methylguanine under each regimen were measured and compared. With the eleven-injection regimen there was a build up of O6-methylguanine in the DNA of the susceptible organ, the kidney, whilst in the liver virtually no accumulation was detected. Under the prolonged, low concentration regimen the liver, in spite of its susceptibility to the carcinogen did not accumulate O6-methylguanine. The results are discussed in terms of the hypothesis that production of O6-methylguanine and its persistence in the DNA of the target organ are responsible for the carcinogenic action of dimethylnitrosamine. 相似文献
14.
Prolactin is an important growth modulatory hormone in fetal and adult tissues. It stimulates DNA synthesis and enzymatic markers of the G1 phase of cell cycle in rat liver and other tissues. In this study the effects of prolactin on 5-methyl cytosine content in liver and kidney of rats was studied using HPLC. Prolactin treatment caused hypomethylation of DNA in the liver and kidney of immature rats at 48 h after treatment and the effect remained even at 72 h. Prolactin also caused hypomethylation of DNA in the kidney and liver of adult rats at 48 h after treatment. These results indicate that prolactin probably regulates DNA methylation in the liver and kidney of immature and adult rats. 相似文献
15.
In the present study, we investigated the protective effect of Bacopa monniera, an indigenous Ayurvedic medicinal plant in India, against morphine-induced liver and kidney toxicity in rats. Morphine intoxicated rats received 10-160 mg/kg body weight of morphine hydrochloride intraperitoneally for 21 days. Bacopa monniera Extract (BME) pretreated rats were administered with BME (40 mg/kg) orally once a day 2 h before the injection of morphine for 21 days. Pretreatment with BME has shown to possess a significant protective effect against morphine-induced liver and kidney functions in terms of serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, alkaline phosphatase, lactate dehydrogenases and gamma-glutamyl transferase activities and urea, creatinine and uric acid level respectively. Histopathological changes of liver and kidney were also in accordance with the biochemical findings. The results of this study indicate that Bacopa monniera extract exerted a protection against morphine-induced liver and kidney toxicity. 相似文献
16.
VITAMIN B12 AND THE MACROMOLECULAR COMPOSITION OF EUGLENA : III. Effect of Cycloheximide on the Recovery from B12-Induced Unbalanced Growth 总被引:1,自引:0,他引:1
下载免费PDF全文
![点击此处可从《The Journal of cell biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
George H. Goetz Pamela L. Johnston Kathy Dobrosielski-Vergona Edgar F. Carell 《The Journal of cell biology》1974,62(3):672-678
When cycloheximide is added to (B12)-deficient cultures before or after replenishment of the cells with B12, reversion of these cells is inhibited. This inhibition is not caused by interference of the inhibitor in the uptake of B12 as measured by division kinetics. Cycloheximide does not inhibit the initial increase in the rate of DNA synthesis caused by B12 replenishment, but within 30–45 min the rate decreases and DNA synthesis ceases. Cycloheximide added to replenished deficient cells after completion of DNA duplication inhibits cell division. The total cellular protein and RNA in replenished cells treated with cycloheximide does not change. B12 added to deficient cells does not stimulate the incorporation of [14C]leucine into protein during resumption and completion of DNA duplication. However, there is a large increase in [14C]leucine incorporation into the protein of these cells soon after completion of DNA duplication and before resumption of cell division. The addition of cycloheximide to B12-replenished or to nonreplenished deficient cells rapidly inhibits the incorporation. We suggest that the addition of B12 accelerates the rate of DNA synthesis in the deficient cells and that possibly no new protein synthesis is required except for mitosis. However, protein synthesis is needed for continuous DNA synthesis. 相似文献
17.
We studied DNA metabolism (synthesis and degradation) in brain to investigate the effect of hyperphenylalaninemia induced in rats by treatment with PCPA or MPA plus PHE during suckling (4th–20th days of postnatal age) on cell proliferation and naturally occurring cell death. The incorporation of14C in DNA as percent of total radioactivity in the tissue, 30 min after administration of [14C]thymidine served as a measure of DNA synthesis in vivo, and the amount of radioactivity recovered in DNA as percent of total14C in the tissues of 21 day old rats, injected with [14C]thymidine on 2nd day after birth, indicated the turnover (degradation) of DNA. The results showed that the DNA content of cerebellum as well as cerebrum was reduced by treatment with PCPA plus PHE, while treatment with MPA plus PHE had no effect on DNA content in cerebellum but reduced the levels in cerebrum. Treatment with PCPA or MPA plus PHE reduced the synthesis of DNA in cerebrum of 11 day old rats but not in 21 day old rats, and the treatments did not affect DNA synthesis in cerebellum of either 11 or 21 day old rats. The turnover (degradation) of DNA was increased in both cerebellum and cerebrum from rats treated with PCPA plus PHE but MPA plus PHE treatment did not alter the DNA turnover either in cerebellu or in cerebrum. The activity of acid DNase was reduced in both cerebellum and cerebrum from 11 as well as 21 day old rats treated with PCPA plus PHE, but the enzyme activity was not altered in the tissues from rats of both ages treated with MPA plus PHE. The data thus indicate that in rats treated with PCPA plus PHE the reduction in cerebral DNA levels occurs due to reduced synthesis and/or increased turnover (degradation) of DNA but that the reduction in cerebellar DNA may occur only as a result of increased turnover (degradation), and that in rats treated with MPA plus PHE the reduction in cerebral DNA must occur due to reduced synthesis. This suggests that treatment of rats with PCPA plus PHE during suckling inhibits cell proliferation and/or increases naturally occurring cell death in both cerebellum and cerebrum while treatment with MPA plus PHE inhibits only cell proliferation and in cerebrum alone. 相似文献
18.
Samantha A. Beck 《Developmental biology》2010,339(2):320-2788
Polycomb group (PcG) genes are required for heritable silencing of target genes. Many PcG mutants have chromatin bridges and other mitotic defects in early embryos. These phenotypes can arise from defects in S phase or mitosis, so the phenotype does not show when PcG proteins act in cell cycle regulation. We analyzed the cell cycle role of the proximal subunit of Polyhomeotic (PhP) in Drosophila. Time-lapse imaging reveals that chromatin bridges formed during mitosis are able to resolve but sometimes result in chromosome breakage. Chromosome bridging is also observed in canonical cell cycles occurring in larval brains and is therefore not unique to the rapid embryonic cycles. PhP colocalizes with chromatin in S phase but not in mitosis in early embryos, indicating a direct role in DNA synthesis. Time lapse imaging of php mutants reveals an acceleration of S phase, showing that php regulates S phase length. Like php mutations, mutations in DNA damage checkpoints result in S phase acceleration. Consistent with this model, mutations in ph do not affect DNA synthesis rates, but exhibit impaired ability to block cell cycle progression following exposure to gamma-rays. Our data show that the mitotic defects of php are caused by defects in the DNA damage response that occurs after DNA replication in S phase, and we propose that PhP has a direct role in DNA damage repair. 相似文献
19.
1. The nucleic acid metabolism in the pyridoxine-deficient rat has been investigated through studies on the incorporation of radioactivity from various isotopically labelled compounds into liver and spleen DNA and RNA. 2. In pyridoxine deficiency, the incorporation of radioactivity from sodium [14C]formate was apparently increased. The magnitude of this effect on incorporation into liver RNA and DNA and spleen RNA was approximately the same. The incorporation into spleen DNA was enhanced to a much greater degree. Administration of pyridoxine 24hr. before the rats were killed reversed the changes in incorporation of radioactivity from [14C]formate. 3. In pyridoxine deficiency, the incorporation of radioactivity from dl-[3-14C]serine, [8-14C]adenine, [Me-3H]thymidine and [2-14C]deoxyuridine was decreased. The incorporation of radioactivity from l-[Me-14C]methionine was not affected. No noteworthy differences in the effect of pyridoxine deficiency on the incorporation of radioactivity from dl-[3-14C]serine into DNA and RNA were observed, whereas the effect of the deficiency on the incorporation of radioactivity from [8-14C]adenine into spleen DNA was somewhat greater than that into spleen RNA. Administration of pyridoxine 24hr. before the rats were killed reversed the changes in incorporation of radioactivity from [3-14C]serine and [8-14C]adenine. 4. The adverse effects of pyridoxine deficiency on the biosynthesis of nucleic acids and cell multiplication are discussed in relation to the role of pyridoxal phosphate in the production of C1 units via the serine-hydroxymethylase reaction. 相似文献
20.
When a single dose of urethan was injected into the peritoneal cavity of rats immediately after partial hepatectomy, DNA synthesis was delayed by 12 h. The induction of ornithine decarboxylase which was induced biphasically following partial hepatectomy was also reduced and delayed by 14–15 h by the administration of urethan. S-Adenosylmethionine decarboxylase activity in urethan-treated rat liver at 20 h and 29 h after operation was significantly lower than that of untreated animals. This enzyme activity was shown to increase thereafter, reaching a higher level than in untreated rats at 37–42 h. Hepatic spermidine content changed biphasically in a manner similar to DNA synthesis. These results suggest that the activities of ornithine decarboxylase and S-adenosylmethionine decarboxylase may correlate with DNA synthesis and that an increase of spermidine concentration is necessary to DNA synthesis. 相似文献