Effect of omega-3 polyunsaturated fatty acids on activity of glutathione-dependent enzymes in the liver cytosol and blood erythrocytes in rats with experimental chronic bronchitis and in the norm

The influence of omega-3 polyunsaturated fatty acids (omega-3 PUFA) on the activity of glutathione reductase, glutathione transferase and glutathione peroxidase in the liver cytosole and red blood cells of normal rats and animals with experimental chronic bronchitis. omega-3 PUFA ("Tekom" medication) activate glutathione reductase of liver cytosole and glutathionperoxidase in the red blood cells in rats. In the rats with chronic inflammatory process in bronchia omega-3 PUFA corrects the glutathione-dependent systems of detoxication. Effects were more expressed in the liver cytosole in comparison with the red blood cells. The using of omega-3 PUFA as a means for treatment and prophylaxis was more effective than for treatment only.     

Destructive and restorative processes of the liver in rats under intensive physical loading

The ultrastructure of rat liver cells after running exercise was investigated. When rats were trained for a month and sacrificed immediately after the last exercise it was revealed that the number of liver cells mitochondria increased, but many of them had alterations: mitochondria became swollen, had lucid matrix. There were some variations in degree of alterations between different mitochondria: a) in the same hepatocyte, b) in different hepatocytes of the same animal, that was connected with individual sensitivity of organelles on the levels of the cell and of the organ. Rough endoplasmic reticulum bore few ribosomes. Glycogen was absent. There were abundant vesicles of smooth endoplasmic reticulum, autophagic vacuoles and peroxisomes in the liver cell cytoplasm. Adaptation of rat liver to the exercise programme becomes evident by 1.5 month of exercise. Mitochondria and rough endoplasmic reticulum were numerous and of normal structure. There were many peroxisomes and glycogen granules in the cytoplasm of hepatocyte. The presence of large autophagic vacuoles in the cytoplasm of some hepatocytes were obviously connected with more rapid destruction of some organelles, than in control.     

Influence of proliferation on DNA repair rates in liver

To test the hypothesis that the proliferative status of a mammalian cell determines the rate of removal of oxidative DNA damage, pre- and posthepatectomized livers in adult male Fisher 344 rats were irradiated in situ with 15.5 Gy of 137Cs-gamma-rays. At 10 and 45 min after irradiation, the livers were removed and dissociated into single cell suspensions, and the DNA damage in the isolated quiescent or proliferative liver cells was assayed by alkaline elution. Proliferative liver cells irradiated 20-24 h or 29-31 h after hepatectomy repaired their DNA damage faster than quiescent liver cells. A corresponding increase in the accessibility of the DNA to digestion by m. nuclease was observed for the post-hepatectomized liver cells. These data suggest that proliferative status is a major determinant of the rate of DNA repair in rat liver.     

Lipid metabolism and structure-activity features of the endoplasmic reticulum membrane in regenerating rat liver]

The relationship between the neutral lipid and phospholipid metabolism and some structure-function peculiarities of regenerating rat liver endoplasmic reticulum membranes (13 hours after surgery, i.e., corresponding to the G1-period of the cell cycle) was studied. There was an increase in the degree of the endoplasmic reticulum membrane development and the nonesterified fatty acid (NFA) and triglyceride (TG) content in regenerating rat liver microsomes. The relative specific radioactivity of neutral lipid and phospholipid fractions in regenerating rat liver microsomes was lower than in control animals, presumably due to the high rate of the microsomal lipid exchange in the regenerating liver with other cell organelles. The changes in the lipid content and rate of their metabolism in the regenerating rat liver were associated with the increase in the membrane microviscosity and the decrease in the activity of the membrane-bound enzyme (glucose-6-phosphatase). The differences in the time-dependent changes in the synthesis and metabolism of lipids in the NFA and TG fractions may be regarded as an endogenous factor determining the structure-function peculiarities of endoplasmic reticulum membranes.     

Effects of ubiquinone-9 on lipids of nuclei and chromatin of the rat liver under continuous irradiation]

The influence of continuous gamma irradiation on the lipids of nuclei and chromatin of rat liver at a dose-rate of 0,129 Gy/day for 155 days (a total dose of 20 Gy) and by feeding of ubiquinone-9 has been studied. The amount of phosphatidylcholine with phosphatidylserine and phosphatidyl-ethanolamine in liver nuclei of irradiated rats was found to increase. Ubiquinone-9 had a normalizing effect. A decrease of cardiolipin was observed in the liver chromatin of irradiated rats. The amount of free fatty acids had a tendency to decrease in homogenate, nuclei and liver chromatin of irradiated rats. Ubiquinone was found to increase the amount of free fatty acids up to the control level. The amount of cholesterol in nuclei was increased after irradiation and that in chromatin tended to rise. Ubiquinone-9 significantly decreased the amount of cholesterol in nuclei and chromatin of irradiated rats.     

Effect of tocopherol and tocopherol quinone complexes with proteins on activity of leukotriene B4 lipoxygenase

While estimating the effect of tocopherol, tocopherylquinone and their complexes with the tocopherol-binding proteins from the rat liver cytosole on arachidonate 5-lipoxygenase from peritoneal-lymphocytes and soybean linoleate-5-lipoxygenase alpha-tocopherol and especially its complex with tocopherol-binding protein was defined to inhibit the activity of both vegetative- and animal nature-lipoxygenase.     

Induction of a Peroxisomal Malate Dehydrogenase Isoform in Liver of Starved Rats

The influence of starvation on malate dehydrogenase (MDH) in rat liver was investigated. Native electrophoresis revealed two MDH isoforms in non-starved rats and three isoenzymes in starved rats. After sucrose density gradient centrifugation of cell organelles from liver, MDH activity was detected in the mitochondrial and cytosolic fractions from non-starved rats. However, additional activity was found in the peroxisomal fraction from starved rats. The latter was identified as the electrophoretically new isoform in starved animals. The three isoforms of malate dehydrogenase from hepatocytes were separated and partially purified by chromatography on DEAE-Toyopearl. Several kinetic and regulatory properties of the three isoforms were rather similar. It is suggested that the newly expressed isoform of MDH operates in the glyoxylate cycle of liver peroxisomes of food-starved animals.     

Antiapoptotic effect of l-carnitine on testicular irradiation in rats

We evaluated the effects of l-carnitine on apoptosis of germ cells in the rat testis following irradiation. Male Wistar rats were divided into three groups. Control group received sham irradiation plus physiological saline. Radiotherapy group received scrotal gamma-irradiation of 10 Gy as a single dose plus physiological saline. Radiotherapy + l-carnitine group received scrotal irradiation plus 200 mg/kg intraperitoneally l-carnitine. Twenty-four hours post-irradiation, the rats were sacrificed and testes were harvested. Testicular damage was examined by light and electron microscopy, and germ cell apoptosis was determined by terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate in situ nick end-labeling (TUNEL) technique. Morphologically, examination of irradiated testis revealed presence of disorganization and desquamation of germinal cells and the reduction in sperm count in seminiferous tubule lumen. Under electron microscopy, the morphological signs of apoptosis were frequently detected in spermatogonia. Apoptotic spermatogonia showed the marginal condensation of chromatin onto the nuclear lamina, nucleus and cytoplasm shrinkage and still functioning cell organelles. TUNEL-positive cells were significantly more numerous in irradiated rats than in control rats. l-carnitine treatment significantly attenuated the radiation-induced morphological changes and germ cell apoptosis in the irradiated rat testis. In conclusion, these results suggested that l-carnitine supplementation during the radiotherapy may be beneficial for spermatogenesis following testicular irradiation by decreasing germ cell apoptosis.     

Changes of transferrin-free iron uptake by bone marrow erythroblasts in strenuously exercised rats

The effects of strenuous exercise on transferrin-free iron (Fe II) uptake by bone marrow erythroblasts in rats were investigated. Female Sprague-Dawley rats were randomly assigned to one of six groups, three of which underwent 3, 6, or 12 months of strenuous exercise (swimming 2 hr/day, 5 days/week) or their corresponding three control groups. At the end of experiments, bone marrow erythroblasts were isolated for Fe II uptake assay in vitro. It was found that the amounts of iron uptake into cytosole and stroma of the cells of rats in the groups undergoing 3 and 12 months of exercise did not differ from those in their corresponding sedentary groups. In addition, analysis of nonspecific and specific Fe II uptake by cytosole and stroma did not display any significant difference between the exercise and corresponding sedentary groups. However, the amount of Fe II uptake into cytosole and stroma was significantly increased in rats that exercised for 6 months compared with the corresponding controls. Nonspecific iron uptake into stroma was significantly higher in the exercise group than in the sedentary group (0.120 +/- 0.018 vs. 0.049 +/- 0.006 pM/10(6) cells, P < 0.01). The V(max) of the specific iron uptake into stroma was significantly higher (0.326 +/- 0.024 vs. 0.238 +/- 0.037 pM/10(6) cells, P < 0.05) and the K(m) of iron uptake into cytosol lower (0.08 +/- 0.01 vs. 0.21 +/- 0.03 microM, P < 0.001) in the exercise groups than in the control groups. These results indicate that 6 months of strenuous exercise could significantly increase Fe II uptake by the cells, probably by affecting the number and/or affinity of the putative iron carrier and the permeability to iron of cell membrane. The increased ability of cell-free iron accumulation in exercise might be a self-protective mechanism for body cells from the free iron-induced free radical reaction in addition to providing more iron for cell heme synthesis.     

Comparative studies on lipid peroxidation of microsomes and mitochondria obtained from different rat tissues: effect of retinyl palmitate

The effect of retinyl palmitate on the polyunsaturated fatty-acid composition, chemiluminescence and peroxidizability index of microsomes and mitochondria obtained from rat liver, kidney, brain, lung and heart, was studied. After incubation of microsomes and mitochondria in an ascorbate Fe++ system (120 min at 37 degrees C) it was observed that the total cpm/mg protein originated from light emission: chemiluminescence was lower in liver microsomes, mitochondria and kidney microsomes in the vitamin A group than in the control group. In mitochondria obtained from control rats, the most sensitive fatty acids for peroxidation were arachidonic acid C20:4 n6 in liver and docosahexaenoic acid C22:6 n3 in kidney and brain. In microsomes obtained from control rats, the most sensitive fatty acids for peroxidation were linoleic acid C18:2 n6 and C20:4 n6 in liver and C22:6 n3 in kidney. Changes in the most polyunsaturated fatty acids were not observed in organelles obtained from lung and heart. As a consequence the peroxidizability index, a parameter based on the maximal rate of oxidation of fatty acids, showed significant changes in liver, kidney and brain mitochondria, while in microsomes changes were significant in liver and kidney. These changes were less pronounced in membranes derived from rats receiving vitamin A. Our results confirm and extend previous observations that indicated that vitamin A may act as an antioxidant protecting membranes from deleterious effects.     

蒌蒿对镉的富集特征及亚细胞分布特点

以镉(Cd)富集植物蒌蒿(Artemisia selengensis)为试材, 采用超声波细胞破碎处理和超速离心的方法, 对蒌蒿根和叶中亚细胞水平的Cd分布状况进行研究, 同时测定Cd在蒌蒿不同器官中的富集效果。结果表明, 在30 mg·kg^–1Cd胁迫下, 蒌蒿叶片中Cd的富集浓度是根和茎中的2–3倍, 但因叶片所占植株的生物量比例较小, 其对Cd的积累量远小于茎和根; Cd在蒌蒿叶片细胞壁、胞液和细胞器中含量比为16:5:1。细胞壁固定是叶片对Cd的主要防御机制。随着Cd处理浓度的增加, 细胞壁和胞液中的Cd含量大幅上升, 但细胞器中Cd含量仍维持在较低水平。长时间和高浓度的Cd胁迫可使细胞壁解毒机制失活并诱导细胞器中的Cd含量增加, 导致植株死亡。根中液泡的Cd贮存量较大, 解毒效果显著。     

Effect of ubiquinone on phospholipid metabolism in radiation injury

The synthesis and phospholipid content in the liver, intestine and spleen in normal and irradiated rats administered ubiquinone-9 were studied with the use of 3H-serine. Ubiquinone markedly activated decarboxylation of phosphatidylserine and suppressed transformation of phosphatidylethanolamine to phosphatidylcholine in rat liver and spleen. The effect was also observed in the organs of irradiated animals. In rat intestine, administration of ubiquinone normalized a sharp gamma-irradiation-induced inhibition of transformation of phosphatidylcholine from phosphatidylethanolamine. The catabolism of phospholipids under the action of ubiquinone and radiation was inhibited in the liver and, on the contrary, was activated in radiosensitive organs.     

Radioprotective properties of ubiquinones during acute and chronic irradiation of rats

Protective and therapeutic effect of ubiquinone-8 was demonstrated on rats exposed to 8 Gy radiation. There was a 16-20% increase in the survival rate of the irradiated rats as compared with the controls. The per os administration of ubiquinone-8 during a five-month period in conditions of chronic exposure arrested the radiation-induced accumulation of lipids in rat liver and somewhat improved the dynamics of the body mass growth in the exposed rats.     

Studies of the teratogenic potential of exposure of rats to 6000-MHz microwave radiation. I. Morphologic analysis at term

Thirty-six pregnant Wistar strain albino rats were exposed throughout pregnancy to 6000-MHz microwave radiation at a power density level of 35 mW/cm2 or were used as controls. The irradiation did not cause a significant increase in maternal body temperature as measured by a rectal thermocouple. The rats were randomly assigned to one of four groups: home cage control (5), anechoic chamber control (10), sham-irradiated concurrent control (10), and irradiated (11). All animals were killed on the 22nd day of gestation, and maternal tissues were removed and weighed and maternal blood samples were taken. The 384 resultant fetuses and their placentas were individually weighed, fixed, and dissected to determine normality. Teratologic evaluation included the following parameters: maternal weight and weight gain; mean litter size; maternal organ weight and organ weight/body weight ratios; body weight ratios of brain, liver, kidneys, and ovaries; maternal peripheral blood parameters including hematocrit, hemoglobin, and white cell counts; number of resorptions and resorption rate; number of abnormalities and abnormality rate; mean term fetal weight. The irradiated fetuses exhibited slight but statistically significant growth retardation at term. Term maternal monocyte count was also significantly depressed. No other parameters differed between the control groups and the irradiated group.     

The proteinase activity in the cell nuclei of rats following exposure to gamma radiation and methylnitrosourea

Activity of nuclear proteinases in blood and liver cells of rats exposed to whole-body gamma-irradiation (10 Gy) has been comparatively studied by the capacity of splitting the casein substrate. Proteinase activity in nuclei of irradiated rat leukocytes was shown to increase by 2.5 times and to gradually decrease after 48 h reaching 150-160% as compared to the control. Two hours following a single injection of methyl nitrosourea the alteration in the activity of proteinases in nuclei of rat hepatocytes and leukocytes was different from the alteration of this index after gamma-irradiation.     

Effect of neonatal head X-irradiation on growth of costal and tibial cartilage in rats: a histochemical and electron microscopic study

Long-Evans rats were exposed to a single dose of head X-irradiation (600 rads) at 2 days of age. Experimental and sham irradiated rats were sacrificed at 14, 20-21, 23, 41-45, and 70-71 days. Tibial epiphyseal width and the number of cells in the epiphyseal plate were determined. Histochemical and electron microscopic studies were carried out on both costal and epiphyseal cartilage. Histochemical techniques revealed a reduction in chondroitin sulfate at 14 days in both costal and epiphyseal cartilage of X-irradiated rats. Epiphyseal cartilage demonstrated recovery subsequently, and this was followed by a normal decrease of chondroitin sulfate with increasing age, but costal cartilage did not recover. Collagen synthesis was also reduced in both costal and epiphyseal cartilage, but not as dramatically as chondroitin sulfate. Except for some electron dense cells and reduced scalloping of the cell membrane, costal chondrocytes from irradiated rats did not show major ultrastructural alterations. In contrast, epiphyseal chondrocytes demonstrated radiation induced alterations in organelles, in enhanced glycogen deposition, and in retardation of chondrocyte maturation. Extracellularly in both costal and epiphyseal cartilage of irradiated rats, collagen density and matrix granules were reduced, while calcification of the matrix was enhanced. Beyond 45 days, the effects of irradiation were markedly reduced. Comparisons of the histochemical results with metabolic studies carried out previously in cartilage from the same animals indicated a more direct concordance of the histochemical results with the pattern of physical growth and supported the usefulness of morphologic and histochemical techniques in the analysis of the growth disorder in the head-irradiated rat.     

The effect of dexamethasone on transferrin secretion by cultured fetal rat hepatocytes

Cultured fetal rat hepatocytes derived from 12, 15 and 19-day gestation rats are capable of secreting transferrin. When dexamethasone is added to the medium an increased secretion rate is observed. The changes in secretion rates in control as well as dexamethasone-treated cells during culture have been shown to correlate with the level of mRNA coding for transferrin. Immunocytochemical experiments show that initially all hepatocytes contain transferrin which is localized in the lumina of the perinuclear space, rough endoplasmic reticulum and in the saccules and vesicles of the Golgi apparatus. During culture, particularly in control cells, the intensity of labelling varies from cell to cell. In addition, adjacent cells are observed to label more intensely in different intracellular organelles.     

Organic anion transporters,OAT1 and OAT3, are crucial biopterin transporters involved in bodily distribution of tetrahydrobiopterin and exclusion of its excess

Like mitochondria, peroxisomes produce reactive oxygen species (ROS), compounds which have been implicated to play an important role in many degenerative diseases and aging itself, and an exaggerated ROS production might occur in altered or older organelles. Growing evidence shows that autophagy, a required function in cell housekeeping during fasting, can remove damaged macromolecules, organelles, and membranes selectively. Proliferation of peroxisomes can be enhanced in liver cells by perfluorooctanoic acid (PFOA), which causes a marked increase of the Acyl-CoA oxidase (ACOX) activity and no significant change in urate oxidase (UOX) activity. The administration of antilipolytic drugs to fasted animals was shown to intensify autophagy. Here we tested the hypothesis that autophagy may distinguish and remove older from younger peroxisomes in rat liver. Male Sprague-Dawley rats were given PFOA (150 mg/kg body weight) or vehicle. Animals were sacrificed at different times following PFOA administration, and 3 h after the induction of autophagy with the antilipolytic agent 3,5-dimethyl pyrazole (DMP, 12 mg/kg body weight). The levels of ACOX and UOX activity were measured in the liver tissue. Results showed that autophagy caused a parallel, significant decrease in both enzymes activity in control rats, and that in PFOA-treated rats the effects were different and changed with PFOA time administration. Changes are compatible with the hypothesis that newly formed ACOX-rich peroxisomes are resistant to pexophagy and that sensitivity to pexophagy increases with increasing peroxisomal “age.” In conclusion, there is indirect evidence supporting the hypothesis that autophagy may recognize and degrade older peroxisomes.     

Non-enzymatic lipid peroxidation of microsomes and mitochondria isolated from liver and heart of pigeon and rat

Studies were carried out to determine the level of ascorbate-Fe2+ dependent lipid peroxidation of mitochondria and microsomes isolated from liver and heart of rat and pigeon. Measurements of chemiluminescence indicate that the lipid peroxidation process was more effective in mitochondria and microsomes from rat liver than in the same organelles obtained from pigeon. In both mitochondria and microsomes from liver of both species a significant decrease of arachidonic acid was observed during peroxidation. The rate C18:2 n6/C20:4 n6 was 4.5 times higher in pigeon than in rat liver. This observation can explain the differences noted when light emission and unsaturation index of both species were analysed. A significant decrease of C18:2 n6 and C20:4 n6 in pigeon liver mitochondria was observed when compared with native organelles whereas in pigeon liver microsomes only C20:4 n6 diminished. In rat liver mitochondria only arachidonic acid C20:4 n6 showed a significant decrease whereas in rat liver microsomes C20:4 n6 and C22:6 n3 decreased significantly. However changes were not observed in the fatty acid profile of mitochondria and microsomes isolated from pigeon heart. In the heart under our peroxidation conditions the fatty acid profile does not appear to be responsible for the different susceptibility to the lipid peroxidation process. The lack of a relationship between fatty acid unsaturation and sensitivity to peroxidation observed in heart suggest that other factor/s may be involved in the protection to lipid peroxidation in microsomes and mitochondria isolated from heart.     

大鼠肝移植术后腹腔细菌感染模型的建立与评价

目的建立大鼠肝移植术后腹腔感染的模型。方法构建DA大鼠到LEW大鼠的肝移植模型,采用腹腔内细菌注射的方法建立感染模型,通过对大鼠肝功、血气、血细胞计数等各项指标的检测对模型进行综合评价。结果肝移植术后5 d注射细菌,大鼠死亡率高,不利后续研究;术后3 d注射细菌,并选定5×105cfu/mL为最终注射浓度,感染后大鼠的7d存活率累计可达到37.5%左右,随之感染的加重,大鼠状态逐渐变差,直肠温度不断升高,WBC计数也随之增加,pH下降,大鼠出现代谢性酸中毒,肝功能损害进行性加重,肝实质的损害重于胆道的损伤,大约在感染5 d左右相继死亡,多器官病理分析表明,大鼠死亡原因为肝损害,不并发肺脏、肾脏损害。结论采用的腹腔内大肠埃希菌注射建立肝移植术后腹腔细菌感染的模型是比较成功的,可用于相关领域的研究。