共查询到20条相似文献,搜索用时 15 毫秒
1.
Bin Zhu Yuewen Gong Pengmin Chen Haojun Zhang Tingting Zhao Ping Li 《Molecular and cellular biochemistry》2014,393(1-2):23-32
DNase I is an endonuclease responsible to destruction of chromatin during apoptosis. However, its role in diabetes is still unclear. With blood samples from our previous study related to type 2 diabetes, we examined the DNase I activity in the serum of these patients and the role of DNase I in the injury of pancreas was further investigated in rats and INS-1 cells. Serum and pancreatic tissues from human and rats were used for the study. Insulin resistance and diabetes were induced by high fat diet and STZ injection, respectively. DNase I activity was determined by radial enzyme-diffusion method. Expressions of DNase I and caspase-3 in pancreas were determined in rat pancreatic tissues and INS-1 cells. Apoptosis of INS-1 cells was determined by both TUNEL assay and Flow Cytometry. There was a significant elevation of DNase I activity in serum of patients with type 2 diabetes and rats with STZ injection. Moreover, increase in DNase I expression was observed in the pancreas of diabetic person and rats. Furthermore, high glucose induced both DNase I and caspase-3 expression and at the same time increased apoptosis rate of INS-1 cells. In conclusion, elevated DNase I in diabetes may be related to pancreatic injury and could be one of the causes that induce diabetes. 相似文献
2.
Fujihara J Tabuchi M Inoue T Yasuda T Fujita Y Takeshita H 《Analytical biochemistry》2011,(1):2591-79
Deoxyribonuclease I (DNase I) activity in serum has been shown to be a novel diagnostic marker for the early detection of acute myocardial infarction (AMI). However, the conventional method to measure DNase I activity is time-consuming. In the current study, to develop a rapid assay method for DNase I activity for clinical purposes, a microchip electrophoresis device was used to measure DNase I activity. Because DNase I is an endonuclease that degrades double-stranded DNA endo-nucleolytically to produce oligonucleotides, degradation of the DNA standard caused by DNase I action was detected using microchip electrophoresis. We detected DNase I activity within 10 min. This is the first study to apply microchip electrophoresis for the detection of DNase I activity; furthermore, it seems plausible that reduction of analysis time for DNase I activity could make this novel assay method using microchip electrophoresis applicable in clinical use. 相似文献
3.
The effect of antigen-induced stimulation on acid deoxyribonuclease (DNase) activity in BALB/c mouse lymphoid cells was determined. Increase in acid DNase activity was found in intact spleen cell populations of mice from the second or fourth day after immunization with pneumococcal polysaccharide type III and from the fourth day after immunization with SRBC. DNase determinations performed with spleen cell fractions prepared from SRBC-immunized mice, showed that the rise in the enzyme activity was confined to the fraction containing the antibody-forming cells. The DNase activity was also increased in spleen cell cultures, stimulated with SRBC in vitro. Rise in the activity of this enzyme was also observed in peritoneal cell populations taken from SRBC-immunized mice. This change was maximal on the second day after immunization, when no appreciable increase in DNase activity of spleen cells was yet detected. The results obtained suggest, that acid DNase is an enzyme involved in the proliferative/maturation response to antigenic stimulation. It is a consequence of antigenic stimulation rather than being involved in the process of afferent stimulation. 相似文献
4.
H S Taper J M Jamison J Gilloteaux C A Gwin T Gordon J L Summers 《The journal of histochemistry and cytochemistry》2001,49(1):109-120
Human prostate cancer cells (DU145) implanted into nude mice are deficient in DNase activity. After administration of a vitamin C/vitamin K(3) combination, both alkaline DNase (DNase I) and acid DNase (DNase II) activities were detected in cryosections with a histochemical lead nitrate technique. Alkaline DNase activity appeared 1 hr after vitamin administration, decreased slightly until 2 hr, and disappeared by 8 hr after treatment. Acid DNase activity appeared 2 hr after vitamin administration, reached its highest levels between 4 and 8 hr, and maintained its activity 24 hr after treatment. Methyl green staining indicated that DNase expression was accompanied by a decrease in DNA content of the tumor cells. Microscopic examination of 1-microm sections of the tumors indicated that DNase reactivation and the subsequent degradation of DNA induced multiple forms of tumor cell death, including apoptosis and necrosis. The primary form of vitamin-induced tumor cell death was autoschizis, which is characterized by membrane damage and the progressive loss of cytoplasm through a series of self-excisions. These self-excisions typically continue until the perikaryon consists of an apparently intact nucleus surrounded by a thin rim of cytoplasm that contains damaged organelles. 相似文献
5.
Ludwig S Mannherz HG Schmitt S Schäffer M Zentgraf H Napirei M 《The international journal of biochemistry & cell biology》2009,41(5):1079-1093
Reduction of serum DNASE1 (DNase I) activity is supposed to aggravate anti-nuclear autoimmunity, i.e. Systemic Lupus Erythematosus (SLE) in man and mice. To evaluate the etiology of this reduction, more information is needed about the source(s) and regulation of serum DNASE1. In this work we used male C57BL/6 wild-type (WT) mice to verify that serum Dnase1 activity partly depends on hepatic Dnase1 gene expression. Thus serum and liver Dnase1 activity showed a parallel oscillatory course during 24h, which was accompanied by a phase-shifted fluctuation of the hepatic Dnase1 mRNA content. Performing native PAGE zymography (NPZ) we detected a presumably premature non-sialylated and a mature sialylated hepatic Dnase1 isoform, which both show a parallel circadian fluctuation, indicating continuous secretion of Dnase1. The sialylated form was also detectable in serum. By immunostaining the hepatocytes were identified as the source of hepatic Dnase1 gene expression. After 70% hepatectomy, the serum Dnase1 activity increased markedly due to the occurrence of ischemic hepatocellular necrosis in the vicinity of the surgical suture. Similarly, hepatocellular necrosis induced by injection of streptolysin-O (SLO) into the liver led to a rapid parallel increase of Dnase1 and of aspartate- and alanine aminotransferase (AST/ALT) in serum. Subsequent to hepatectomy, Dnase1 gene expression was up-regulated in the regenerating liver most likely leading to an enhanced serum Dnase1 level until complete regeneration. These data demonstrate that serum Dnase1 at least partly originates from the liver and hint to the possibility that natural as well as pathological hepatic conditions influence its activity. 相似文献
6.
7.
Immunization of rabbits with DNase I leads to the production of antiidiotypic Abs with DNase activity. It is not known at present whether antiidiotypic Abs against DNA-hydrolyzing enzymes can possess RNase activity. Here we show that immunization of healthy rabbits with bovine DNase I produces IgGs with intrinsic DNase and RNase activities. Electrophoretically and immunologically homogeneous polyclonal IgGs were obtained by sequential chromatography of the immune sera on Protein A-Sepharose and gel filtration. Affinity chromatography on DNA cellulose using elution of Abs with different concentrations of NaCl and an acidic buffer separated catalytic IgGs into four Ab subfractions, three of which demonstrated only DNase activity while one subfraction hydrolyzed RNA faster than DNA. The serum of patients with many different autoimmune (AI) diseases contains small fractions of antibodies (Abs) interacting with immobilized DNA, which possess both DNase and RNase activities. Our data suggest that a fraction of abzymes from AI patients hydrolyzing both DNA and RNA can contain a subfraction of Abs against DNase I. 相似文献
8.
9.
We describe here the characterization of the so far identified human DNase I family DNases, DNase I, DNase X, DNase gamma, and DNAS1L2. The DNase I family genes are found to be expressed with different tissue specificities and suggested to play unique physiological roles. All the recombinant DNases are shown to be Ca(2+)/Mg(2+)-dependent endonucleases and catalyze DNA hydrolysis to produce 3'-OH/5'-P ends. High activities for DNase I, DNase X, and DNase gamma are observed under neutral conditions, whereas DNAS1L2 shows its maximum activity at acidic pH. These enzymes have also some other peculiarities: different sensitivities to G-actin, aurintricarboxylic acid, and metal ions are observed. Using a transient expression system in HeLa S3 cells, the possible involvement of the DNases in apoptosis was examined. The ectopic expression of each DNase has no toxic effect on the host cells; however, extensive DNA fragmentation is observed only in DNase gamma-transfected cells after the induction of apoptosis. Furthermore, DNase gamma is revealed to be located at the perinuclear region in living cells, and to translocate into the nucleus during apoptosis. Our results demonstrate that DNase I, DNase X, DNase gamma, and DNAS1L2 have similar but unique endonuclease activities, and that among DNase I family DNases, DNase gamma is capable of producing apoptotic DNA fragmentation in mammalian cells. 相似文献
10.
Changes in haematopoietic progenitor colony differentiation and proliferation and the production of different abzymes in EAE mice treated with DNA
下载免费PDF全文
![点击此处可从《Journal of cellular and molecular medicine》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Kseniya S. Aulova Ludmila B. Toporkova Julia A. Lopatnikova Alina A. Alshevskaya Sergei V. Sennikov Valentina N. Buneva Thomas Budde Sven G. Meuth Nelly A. Popova Irina A. Orlovskaya Georgy A. Nevinsky 《Journal of cellular and molecular medicine》2017,21(12):3795-3809
Immunization of experimental autoimmune encephalomyelitis (EAE)‐prone C57BL/6 mice with MOG35‐55 (a model used to study aspects of human multiple sclerosis) is known to lead to the production of various abzymes. The production of catalytic IgGs that can efficiently hydrolyse myelin basic protein (MBP), MOG and DNA is associated with changes in the profile of differentiation and level of proliferation of mice bone marrow haematopoietic stem cells (HSCs). As MOG simulates the production of abzymes with high DNase activity, we compared the effects of DNA and MOG immunization on EAE‐prone mice. In contrast to MOG, immunization with DNA leads to a suppression of proteinuria, a decrease in the concentrations of antibodies to MOG and DNA and a reduction in abzyme production. Immunization with DNA only resulted in a significant increase in DNase activity over 40 days where it became 122‐fold higher than before immunization, and fivefold higher when comparing to the maximal activity obtained after MOG treatment. DNA and MOG immunization had different effects on the differentiation profiles of HSCs, lymphocyte proliferation, and the level of apoptosis in bone marrow and other organs of mice. The data indicate that for C57BL/6 mice, DNA may have antagonistic effects with respect to MOG immunization. The usually fast immune response following MOG injection in C57BL/6 mice is strongly delayed after immunization with DNA, which is probably due to a rearrangement of the immune system following the response to DNA. 相似文献
11.
After a long pause, the accumulation of data on the involvement of tumor-specific DNA and extracellular DNA in metastasis has again placed enzymes with deoxyribonuclease activity in the focus of the search for antitumor and antimetastatic drugs. In this work, the ability of bovine pancreatic DNase I to reduce the invasive potential of B16 melanoma has been investigated in vitro and in vivo. It was found that DNase I had a cytotoxic effect on B16 melanoma cells (IC50 ≈ 104 U/mL). At the same time, significantly lower doses of DNase I (102–103 U/mL) inhibited the migratory activity of melanoma cells in vitro, causing a decrease in the distance of cell front migration and in the area of scratch healing 48 h after the enzyme addition, as well as reducing the rate of cell migration. In mice with B16 metastatic melanoma, intramuscular administration of DNase I in the dose range of 0.12–1.20 mg/kg resulted in a two-to threefold decrease in the number of surface lung metastases and caused nonspecific antigenic immune stimulation. 相似文献
12.
Manderson AP Carlucci F Lachmann PJ Lazarus RA Festenstein RJ Cook HT Walport MJ Botto M 《Arthritis research & therapy》2006,8(3):R68-11
Systemic lupus erythematosus (SLE) is characterised by the production of autoantibodies against ubiquitous antigens, especially
nuclear components. Evidence makes it clear that the development of these autoantibodies is an antigen-driven process and
that immune complexes involving DNA-containing antigens play a key role in the disease process. In rodents, DNase I is the
major endonuclease present in saliva, urine and plasma, where it catalyses the hydrolysis of DNA, and impaired DNase function
has been implicated in the pathogenesis of SLE. In this study we have evaluated the effects of transgenic over-expression
of murine DNase I endonucleases in vivo in a mouse model of lupus. We generated transgenic mice having T-cells that express either wild-type DNase I (wt.DNase I)
or a mutant DNase I (ash.DNase I), engineered for three new properties – resistance to inhibition by G-actin, resistance to
inhibition by physiological saline and hyperactivity compared to wild type. By crossing these transgenic mice with a murine
strain that develops SLE we found that, compared to control non-transgenic littermates or wt.DNase I transgenic mice, the
ash.DNase I mutant provided significant protection from the development of anti-single-stranded DNA and anti-histone antibodies,
but not of renal disease. In summary, this is the first study in vivo to directly test the effects of long-term increased expression of DNase I on the development of SLE. Our results are in line
with previous reports on the possible clinical benefits of recombinant DNase I treatment in SLE, and extend them further to
the use of engineered DNase I variants with increased activity and resistance to physiological inhibitors. 相似文献
13.
The effects of halothane on the DNase I activity in an isolated enzyme preparation and in a DNase I-globular (G) actin complex was investigated. DNase I, DNase I-G actin complexes and G actin were exposed to various (0.2–4.0 vol./%) halothane concentrations for 3 h. Thereafter, DNase I was mixed with a DNA solution and the extinction of the acid soluble supernatant of the DNase I assay was determined as a measure of DNase I activity. After 10 min of halothane exposure the DNase I activity is inhibited in direct proportion to halothane concentrations between 0.6 and 4.0 vol/%. After 10 min halothane activates inactive DNase I by inhibiting G actin, an inhibitor of DNase I. G actin, exposed to halothane, does not inhibit the activity of DNase I. The results suggest a mechanism by which halothane may contribute to chromosomal defects and disturbances of DNA metabolism in cells. 相似文献
14.
Quantitation of DNase I sensitivity in Xenopus chromatin containing active and inactive globin, albumin and vitellogenin genes. 总被引:5,自引:3,他引:2
下载免费PDF全文
![点击此处可从《Nucleic acids research》网站下载免费的PDF全文](/ch/ext_images/free.gif)
B K Felber S Gerber-Huber C Meier F E May B Westley R Weber G U Ryffel 《Nucleic acids research》1981,9(11):2455-2474
The disappearance of defined restriction fragments of the beta 1-globin, an albumin and the A1 vitellogenin gene was quantitated after DNase I digestion and expressed by a sensitivity factor defined by a mathematical model. Analysis of naked DNA showed that the gene fragments have similar but not identical sensitivity factors. DNase I digestion of chromatin revealed for the same gene fragments sensitivity factors differing over a much wilder range. This is correlated to the activity of the genes analyzed: the beta 1-globin gene fragment is more sensitive to DNase I in chromatin of erythrocytes compared to hepatocytes whereas the albumin gene fragment is more sensitive to DNase I in chromatin of hepatocytes. The A1 vitellogenin gene has the same DNase I sensitivity in both cell types. Comparing the DNase I sensitivity of the three genes in their inactive state we suggest that different chromatin conformations may exist for inactive genes. 相似文献
15.
Effects of isoflurane on the DNase I activity in an isolated enzyme preparation and in the DNase I-globular (G) actin complex were investigated. DNase I, DNase I-G actin complex, and G actin were exposed to various (0.2-4.0 vol%) isoflurane concentrations for 180 min. Thereafter, DNase I activity was determined. DNase I activity was inhibited in relation to time and concentration of isoflurane exposure. At concentrations ranging from 0.2 to 1.0 vol% of isoflurane inactive DNase I was activated in the DNase I-G actin complex. The DNase I inhibitor G actin showed a reduced capability to inhibit DNase I following isoflurane exposure. Albumin can inhibit the DNase I inactivation possibly by competition in the reactions between DNase I/albumin and isoflurane. After exposure to isoflurane the absorption maximum of DNase I was identical with the absorption maximum of heat-denatured DNase I. The results suggest a mechanism by which isoflurane may affect DNA in an indirect way at concentrations to which the patient is exposed during clinical anesthesia. 相似文献
16.
Kaneko Y Takeshita H Mogi K Nakajima T Yasuda T Itoi M Kuwano H Kishi K 《Journal of biochemistry》2003,134(5):711-718
The DNase I from canine pancreas was purified 260-fold to electrophoretic homogeneity with a 35% yield using three-step column chromatography. The activity of the purified enzyme was completely inhibited by 20 mM EDTA, an antibody specific to the purified enzyme and G-actin. A 1,373-bp cDNA encoding canine DNase I was constructed from the total canine pancreatic RNA using a rapid amplification of cDNA ends method, followed by sequencing. The mature canine DNase I protein was found to consist of 262 amino acids. A survey of DNase I in 13 different canine tissues revealed the highest levels of both DNase I enzyme activity and gene expression in the pancreas; therefore, the canine DNase I is of the pancreatic type. Phylogenetic and sequence identity analyses, studies of immunological properties and the tissue-distribution patterns of DNase I indicated that the canine enzyme is more closely related to the human DNase I than to other mammalian DNases I. Therefore, canine DNase I is found to be one of the best substitutes in studies of human DNase I. 相似文献
17.
Carbohydrate metabolic disorders were investigated by means of enzyme activities in mice (ddYS) injected intraperitoneally with endotoxin from Salmonella typhimurium. The mice exhibited hyperglycemia 2 hr after administration of endotoxin and hypoglycemia at 18 hr. Activity of hepatic phosphorylase in the endotoxin-poisoned mice at 2 hr was slightly higher than that in the control mice, whereas the level of this activity was not significantly different from that in the controls after 18 hr. Glucose-6-phosphatase activity in the poisoned mice increased by 2 hr after injection, but decreased by 18 hr. The blood lactate level in the poisoned mice transiently decreased until 3 hr after injection, but the mice exhibited a marked lactacidemia by 8–24 hr. The time course of lactate dehydrogenase (LDH) activity in various tissues was examined in mice injected with endotoxin. The activity of hepatic LDH declined to about two-thirds of that of the control mice after 16 hr, and was restored to the normal level by 48 hr. LDH in the cardiac muscle was markedly activated (by about 37%) in the early period (3–6 hr) after administration of endotoxin, and this activity gradually declined. However, the activity of LDH in the skeletal muscle showed a tendency similar to the rise and fall of the levels of blood lactate, and was restored to the normal value at 72 hr after injection. On the other hand, the serum LDH activity in the poisoned mice increased about 1.75-fold by 16 hr after injection. Mice injected with endotoxin exhibited a leakage of the isozymes LDH 3 and 5, but the origin of the leakage is uncertain. Similar elevation in the activities of transaminases (GPT and GOT) and malate dehydrogenase was found in the mouse serum at 16 hr after injection of endotoxin. 相似文献
18.
L1210 leukemia cell cytosol was analysed for the presence of DNase I activity. No free activity was determined in crude cytosol. DNase I enzyme was found to occur in a latent form bound to cytoplasmic actin. DNase-actin complex was partially isolated by Sephadex filtration and DNase I-like activity was demonstrated after SDS gel electrophoresis of the complex and enzyme renaturation. The results were compared with those for synthetic complex of pancreatic bovine DNase I and chicken muscle actin. 相似文献
19.
Uwayama J Hirayama A Yanagawa T Warabi E Sugimoto R Itoh K Yamamoto M Yoshida H Koyama A Ishii T 《Biochemical and biophysical research communications》2006,339(1):226-231
Peroxiredoxin I (Prx I) is a key cytoplasmic peroxidase that reduces intracellular hydroperoxides in concert with thioredoxin. To study the role of tissue Prx I in protection from oxidative stress, we generated Prx I-/- mice by gene trapping. We then evaluated the acute-phase tissue damage caused by ferric-nitrilotriacetate (Fe-NTA). Increases in serum aspartate aminotransferase and alanine aminotransferase levels were significantly greater in Prx I-/- than wild-type mice, 4 and 12 h after the injection of Fe-NTA. Using real-time EPR imaging, we examined the reduction of the stable paramagnetic nitroxyl radical 3-carbamoyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl in vivo, and found that the half-life of this spin probe in the liver and kidney was significantly prolonged in the Prx I-/- mice. These results demonstrate that Prx I-/- mice have less reducing activity and are more susceptible to the damage mediated by reactive oxygen species in vivo than wild-type mice. 相似文献
20.
Gary G. Meadows Rokia M. Abdallah Jean R. Starkey Cheryl J. Aslakson 《Cancer immunology, immunotherapy : CII》1988,26(1):67-73
Summary The effect of dietary tyrosine and phenylalanine restriction on splenic natural killer (NK) cell activity was studied in tumor-free B6D2F1 and NIH nude mice and in B16 bladder-6 (BL6) melanoma-bearing B6D2F1 mice. This dietary restriction was found to suppress the naturally elevated NK-cell activity of nude mice and to induce a specific lymphocytopenia in B6D2F1 mice fed the restricted diet for a prolonged period. Baseline NK-cell activity was significantly lower in tumor-free B6D2F1 mice fed a diet restricted in tyrosine and phenylalanine (restricted diet) than in tumor-free mice fed a basal diet. Similar kinetics of activation after a single i.p. injection of 100 g of polyinosinic:polycytidylic acid (poly I:C) were observed in mice fed both diets. NK-cell activity was not significantly augmented after i.v. inoculation of BL6 melanoma, irrespective of the diet fed; however, it was enhanced in tumor-bearing mice after poly I:C injection. This augmentation was similar to that observed in tumor-free mice. Spleen cells from mice fed either diet were responsive to stimulation of NK-cell activity after in vitro incubation with interleukin-2. These results indicate that dietary restriction of tyrosine and phenylalanine, a potentially useful therapeutic adjunct known to lower NK-cell activity, does not significantly interfere with poly I:C or interleukin-2 induction of NK cells. Our results also demonstrate that, while this dietary restriction causes lymphocytopenia, no effect of the diet could be found on total serum IgG or circulating immune complex levels. 相似文献