维生素C多聚磷酸酯对小鼠肝脏抗氧化物酶基因转录的影响

为研究维生素C多聚磷酸酯对小鼠肝脏脂质抗氧化物酶基因转录的影响,将24只3-4周龄、体重为16-22g的健康雄性小鼠随机分为4组,分别在饵料中添加0、500、2500和5000mg/kg的35%维生素C多聚磷酸酯,喂食4周后取其肝脏,用Trizol法抽提总RNA,利用RT-PCR方法对小鼠肝脏超氧化物歧化酶基因、过氧化氢酶基因和谷胱甘肽过氧化物酶基因的mRNA进行分析。结果表明,维生素C多聚磷酸酯对小鼠肝脏抗氧化物酶基因的转录有显著性影响(P<0·05)。维生素C多聚磷酸酯添加量为2500和5000mg/kg的两组,过氧化氢酶基因的mRNA水平明显高于对照组;维生素C多聚磷酸酯添加量为2500和5000mg/kg的两组超氧化物歧化酶基因的mRNA水平明显高于对照组,其中5000mg/kg组的mRNA水平明显高于其它三组;维生素C多聚磷酸酯添加量为5000mg/kg组,谷胱甘肽过氧化物酶基因的转录活性明显高于其它三组(P<0·05)。研究结果表明:高剂量的维生素C多聚磷酸酯能促进小鼠抗氧化物酶基因的转录活性,但促进不同抗氧化物酶基因转录所需的维生素C多聚磷酸酯的量不同。     

Serum selenium,glutathione peroxidase,lipids, and human liver microsomal enzyme activity

This study evaluated selenium status in relation to lipid peroxidation, liver microsomal function, and serum lipids in humans. Serum selenium concentration, glutathione peroxidase (GSH-Px) activity, liver microsomal enzyme activity, assessed by plasma antipyrine clearance (AP-CL) rate, and serum lipids were determined in 23 healthy subjects in a double-blind placebo-controlled trial of selenium supplementation. The low selenium concentration (74.0±14.2 μg/L, mean±SD) is attributable to the low selenium content of the diet. Subjects with the lowest selenium levels (n=11) had reduced serum GSH-Px activity, AP-CL rate, high-density lipoprotein cholesterol (HDL-C), and total cholesterol (T-C) as compared with subjects with higher selenium concentrations (n=12). Low AP-CL rates were associated with low HDL-C: T-C ratios. Selenium supplementation, 96 μg/d for 2 wk, increased serum selenium, GSH-Px activity, and the HDL-C: T-C ratio. The results suggest that a low serum selenium level is associated with a decrease in liver microsomal enzyme activity and serum HDL-C and T-C concentrations. Selenium supplementation in subjects with low serum selenium may favorably influence relations between serum lipoproteins connected with the development of atherosclerotic vascular disease.     

Deletion of selenoprotein P alters distribution of selenium in the mouse

Selenoprotein P (Se-P) contains most of the selenium in plasma. Its function is not known. Mice with the Se-P gene deleted (Sepp(-/-)) were generated. Two phenotypes were observed: 1) Sepp(-/-) mice lost weight and developed poor motor coordination when fed diets with selenium below 0.1 mg/kg, and 2) male Sepp(-/-) mice had sharply reduced fertility. Weanling male Sepp(+/+), Sepp(+/-), and Sepp(-/-) mice were fed diets for 8 weeks containing <0.02-2 mg selenium/kg. Sepp(+/+) and Sepp(+/-) mice had similar selenium concentrations in all tissues except plasma where a gene-dose effect on Se-P was observed. Liver selenium was unaffected by Se-P deletion except that it increased when dietary selenium was below 0.1 mg/kg. Selenium in other tissues exhibited a continuum of responses to Se-P deletion. Testis selenium was depressed to 19% in mice fed an 0.1 mg selenium/kg diet and did not rise to Sepp(+/+) levels even with a dietary selenium of 2 mg/kg. Brain selenium was depressed to 43%, but feeding 2 mg selenium/kg diet raised it to Sepp(+/+) levels. Kidney was depressed to 76% and reached Sepp(+/+) levels on an 0.25 mg selenium/kg diet. Heart selenium was not affected. These results suggest that the Sepp(-/-) phenotypes were caused by low selenium in testis and brain. They strongly suggest that Se-P from liver provides selenium to several tissues, especially testis and brain. Further, they indicate that transport forms of selenium other than Se-P exist because selenium levels of all tissues except testis responded to increases of dietary selenium in Sepp(-/-) mice.     

Levels and 75Se-labeling of specific proteins as a consequence of dietary selenium concentration in mice and rats

Selenium-labeled proteins (SLP) distinct from glutathione peroxidase (GSH-PX) recently have been purified and partially characterized. Antisera to two SLP, a 56-kDa and a 14-kDa protein, were generated in rabbits and used to examine expression of these proteins as a consequence of dietary selenium concentration (0.02, 0.2, 2.0 ppm) in mice and rats. Additionally, the kinetics of 75Se labeling in plasma, liver, kidney, and mammary gland were examined over a 40-hr time period as a function of dietary selenium concentration. A plasma 57-kDa protein was labeled by 30 min after 75Se injection and reached maximum labeling by 4 hr. The cellular 56-kDa and 14-kDa proteins, as well as GSH-Px, labeled progressively over 40 hr starting between 1 and 4 hr after injection. In general, the 56-kDa and GSH-Px followed similar labeling patterns, whereas the 14-kDa protein was labeled less and was not labeled in discernible quantities until 40 hr. The extent of labeling of all proteins was inversely proportional to the dietary selenium concentration and was probably a reflection of different endogenous selenium body pools. The most important observation was generated by the immunoblot data. The amount of 56-kDa and 14-kDa proteins as detected and measured on immunoblots was not a function of dietary selenium concentration. This result suggests that the synthesis and maintenance of the 56-kDa and 14-kDa proteins are not selenium dependent, a characteristic which distinguishes the two proteins from GSH-Px. The single exception to the above results was the 40% decrease of liver 14-kDa protein concentration in carcinogen-treated rats fed 2.0 ppm of selenium. An organic selenium compound, selenobetaine, did not lead to a decrease under similar conditions. In 15 rat mammary tumors induced by 7,12-dimethylbenzanthracene and analyzed on immunoblots, the SLP-56 was undetected in 5 cases and appeared as two bands (56,000 Da, 50,000 Da) in 10 cases. This latter result raises the possibility that the expression of SLP-56 may be altered in mammary tumors as compared with normal mammary gland.     

The effects of feeding condition and dietary lipid level on lipoprotein lipase gene expression in liver and visceral adipose tissue of red sea bream Pagrus major

The effects of feeding condition and dietary lipid level on lipoprotein lipase (LPL) gene expression in the liver and visceral adipose tissue of red sea bream Pagrus major were investigated by competitive polymerase chain reaction. Not only visceral adipose tissue but also liver of red sea bream showed substantial LPL gene expression. In the liver, starvation (at 48 h post-feeding) drastically stimulated LPL gene expression in the fish-fed low lipid diet, but had no effect in the fish fed high lipid diet. Dietary lipid level did not significantly affect the liver LPL mRNA level under fed condition (at 5 h post-feeding). In the visceral adipose tissue, LPL mRNA number per tissue weight was significantly higher in the fed condition than in the starved condition, irrespective of the dietary lipid levels. Dietary lipid levels did not affect the visceral adipose tissue LPL mRNA levels under fed or starved conditions. Our results demonstrate that both feeding conditions and dietary lipid levels alter the liver LPL mRNA levels, while only the feeding conditions but not dietary lipid levels cause changes in the visceral adipose LPL mRNA level. It was concluded that the liver and visceral adipose LPL gene expression of red sea bream seems to be regulated in a tissue-specific fashion by the nutritional state.     

Effects of chromium supplementation on food energy utilization and the trace-element composition in the liver and heart of glucose-exposed young mice

Essentiality of selenium (Se) for Japanese quail,Coturnix coturnix japonica, was confirmed using a formulated semipurified low-Se diet (basal) (0.05 ppm). Selenium-deficiency symptoms appeared in quails on this diet within 15 d, which corresponded to low levels of hemolysate glutathione peroxidase (GSH-Px) activity. Selenium administration at 0.05 and 2.0 ppm levels resulted in an increase of hemolysate GSH-Px activity by 64 and 116%, respectively, in both short- and long-term experiments. Growth over a 2-mo period increased the hemolysate GSH-Px activity by 120% at each level of dietary Se. A differential response was exhibited by hepatic mitochondrial and soluble GSH-Px activity to Se supplementation, the former increasing progressively with increments of Se at 0.05, 2.0, and 4.0 ppm by 45, 70 and 150%, respectively. The soluble GSH-Px activities of tissues, such as liver, kidney, and testis, and RBC membrane-bound activity remained unchanged in long-term studies at different levels of Se. Replenishment of Se to quails maintained on low-Se diets reflected no change in RBC membrane-bound and liver-soluble GSH-Px activities, although the activity in hemolysate increased consistently with Se. The GSH-Px activity in hemolysate was restored to the levels comparable to those of long-term studies only at Se administration at the 2.0-ppm level. The differential response of mitochondrial and soluble GSH-Px activities to Se and other related observations on mitochondrial functions suggest an additional role for Se in mitochondrial membrane processes and glutathione-related metabolic regulations.     

Selenium-mediated biochemical changes in Japanese quails

The effect of selenium (Se) on collagen characteristics and glutathione peroxidase (GSH-Px) activity in the skin of Japanese quailsCoturnix coturnix japonica fed a formulated, semipurified, low-Se diet (basal) (0.05 ppm) was investigated. The quails exhibited severe Se-deficiency symptoms and significant reduction in skin GSH-Px activity at the end of 30 d. Selenium supplementation at a 2-ppm level restored the normal skin conditions and enhanced skin GSH-Px activity significantly. But a dietary Se level of 0.1 ppm was found to be inadequate in restoring the general skin conditions and GSH-Px activity. A markedly low total collagen content of about 23% was observed in the skin of quails fed the basal diet, compared to 39% of total collagen content in the skin of the 2-ppm Se-supplemented group. Molecular organization of skin collagen of quails on the basal and 0.1-ppm Se diet showed an abundance of monomeric forms with less crosslinks, compared to the presence of polymeric forms with more crosslinks, indicating enhanced stability in the skin collagen of quails on the 2-ppm diet. The delay in the in vitro fibril formation of collagen from the basal and 0.1-ppm Se groups, compared to a relatively faster rate in the case of the 2-ppm Se group, indicates a disturbance in the aggregation phenomenon of collagen. The increase in skin GSH-Px activity and concurrent increase in polymeric collagen on increasing the dietary Se level suggest a possible role for Se in collagen metabolism.     

Effects of dietary selenium and tocopherol on glutathione peroxidase and superoxide dismutase activities in rat phagocytes

Glutathione peroxidase activities (GSH-Px) of peritoneal exudate polymorphonuclear neutrophils, pulmonary alveolar macrophages, and peritoneal exudate macrophages of rats depleted of dietary selenium for four to six weeks were markedly lower than the corresponding activities in rats fed the same diet supplemented with 0.5 ppm selenium as sodium selenite. GSH-Px in phagocytes from selenium-supplemented rats adequate or deficient in tocopherol status did not differ significantly. In selenium deficient animals, the residual GSH-Px of polymorphonuclear neutrophils and peritoneal macrophages, but not of alveolar macrophages were slightly higher in tocopherol-deficient rats than in tocopherol-supplemented animals. Superoxide dismutase activities of each cell type were comparable and were not significantly affected by dietary selenium or tocopherol.