Simultaneous subchronic exposure to selenium and diazinon as possible risk factor for osteoporosis in adult male rats

Background

Osteoporosis and its main health outcome, fragility fractures, are large and escalating health problems. Skeletal damage may be the critical result of low-level prolonged exposure to several xenobiotics in the general population, but the mechanisms of their adverse effects are not clearly understood. The current study was aimed to investigate the possible ability of simultaneous subchronic peroral administration of selenium (Se) and diazinon (DZN) to induce changes in bone of adult male rats.In our study, twenty 1-month-old male Wistar rats were randomly divided into two experimental groups. In the first group, young males were exposed to 5 mg Na₂SeO₃/L and 40 mg of DZN/L in drinking water, for 90 days. Ten 1-month-old males without Se and DZN intoxication served as a control group. At the end of the experiment, macroscopic and microscopic structures of the femurs were analysed using analytical scales, sliding instrument, and polarized light microscopy.

Results

The body weight, femoral length and cortical bone thickness were significantly decreased in rats simultaneously exposed to Se and DZN (P < 0.05). These rats also displayed different microstructure in the middle part of the compact bone where vascular canals expanded into central area of substantia compacta. The canals occurred only near endosteal surfaces in rats from the control group. Additionally, a smaller number of primary and secondary osteons, as well as a few resorption lacunae were observed near endosteal surfaces in rats simultaneously administered to Se and DZN. The resorption lacunae as typical structures of bone resorption manifestation are connected with an early stage of osteoporosis. Histomorphometric analysis revealed that area, perimeter, maximum and minimum diameters of primary osteons’ vascular canals were significantly increased (P < 0.05) in the Se-DZN-exposed rats. On the other hand, all measured variables of Haversian canals and secondary osteons were considerable reduced (P < 0.05) in these rats.

Conclusions

Simultaneous subchronic peroral exposure to Se and DZN induces changes in macroscopic and microscopic structures of the femurs in adult male rats, and also it can be considered as possible risk factor for osteoporosis. The current study contributes to the knowledge on damaging impact of several xenobiotics on the bone.     

硒对STZ诱发糖尿病大鼠骨密度影响的研究初报

比较了在低硒饲料的基础上补充不同形态硒对糖尿病大鼠骨密度的影响。健康SD大鼠 1 2 8只 ,体重 50～ 60 g,随机分为 4组 ,每组 32只 ,雌雄各半 ,其中Ⅰ组为低硒对照组 (饲料含硒量 0 0 37mg/kg) ;Ⅱ组为补充亚硒酸钠组 (饲料含硒量 0 3mg/kg) ;Ⅲ组为补充富硒麦芽组(饲料含硒量 0 3mg/kg) ;Ⅳ组为补充富硒酵母组 (饲料含硒量 0 3mg/kg)。Ⅱ、Ⅲ、Ⅳ组的饲料均是在Ⅰ组低硒饲料的基础上分别添加适量的亚硒酸钠、富硒麦芽、富硒酵母后配制而成。喂饲 5周后 ,Ⅰ、Ⅱ、Ⅲ、Ⅳ组用链脲佐菌素 (streptozotocin ,STZ)按 60mg/kg腹腔注射诱发糖尿病 ,继续饲喂 6周。用单光子吸收法测定各组大鼠的肱骨、股骨、胫骨、桡骨的骨密度。结果表明 :①STZ致糖尿病大鼠的骨密度随时间的延长有逐渐下降的趋势。Ⅱ、Ⅲ、Ⅳ补硒组大鼠可以在一定时间内延缓STZ致糖尿病大鼠的骨密度下降。在此期间骨密度的下降以肱骨出现最早 ,股骨次之 ,桡骨、胫骨再次之。②饲料补硒组糖尿病大鼠的肱骨、股骨中硒的含量较饲料低硒组糖尿病大鼠的肱骨、股骨中硒的含量显著提高。③补充硒的形态 ,即有机硒 (富硒麦芽、富硒酵母 )或无机硒 (亚硒酸钠 )对糖尿病大鼠肱骨、股骨、胫骨、桡骨的骨密度影响差异不显著。     

Early changes of osteochondrosis in medial femoral condyles from rats

Osteochondrosis developed from the early growing process of articular cartilage at the caudal-central region of the medial femoral condyle in rats. Articular cartilage was thick at the region. Mineralization of the matrix in the thick deep zone was incomplete and major parts remained unmineralized. Cavity formation in the mineralized matrix resulting in osteochondrotic lesions was present in the deep zone at 6 weeks of age and was followed by an appearance of viable chondrocytes around it. Osteochondrotic lesions were present from the age of 10 weeks for females and 12 weeks for males. Cavities were expanded and increased in number, and eosinophilic necrotic foci were additionally seen. These changes were extended throughout the deep zone, and viable chondrocytes were also increased in number. The thick deep zone was retained and had no detectable invasions of blood vessels from the subchondral bone. At 20 weeks of age, necrotic areas containing large clefts were present in the basal layer of the thick deep zone and fibrotic lesions were seen beneath them. In normal cases, invasions of blood vessels were seen in the basal layer of the deep zone and also in the cavities of the cartilage; the deep zone was markedly thinned at 20 weeks of age.     

Morphologic changes in chicken cells after in vitro exposure to Mycoplasma gallisepticum

Mycoplasma gallisepticum (MG) was used to expose chicken peripheral blood lymphocytes (PBLs), red blood cells (RBCs), heterophils, and chicken tumor cells (MSB-1 and HD-11 cells). Incubation of PBLs with MG for 3 hr resulted in extensive clumping of lymphocytes. Incubation of the MSB-1 cells with MG also caused clumping of the cells, with many of the cells showing perforations and others showing capping of the surface projections. Incubation of RBCs with MG resulted in an altered cell surface morphology, a decrease in cell size, and perforation. There were no discernible changes on the surface of the heterophils and the HD-11 cells. However, the HD-11 cells appeared to have a decreased ability to attach to the surface of the plastic and to have a decreased ability to respond to chemoattractant fMLP after 24 hr of incubation. These results suggest that, under the conditions used, MG caused certain damage to peripheral blood cells and a significant decrease in chemotactic response in the HD-11 cells.     

Effect of chronic exposure to excess dietary copper and dietary selenium supplementation on liver specimens from rats

OBJECTIVE: To determine the effects of chronic exposure to excess dietary copper (Cu) on liver specimens from rats and the effects of dietary selenium (Se) supplementation in experimental Cu toxicosis. ANIMALS: 60 weanling male Fischer 344 rats. PROCEDURE: Rats were randomly assigned to 4 groups of 15 rats each and fed 1 of the following 4 diets: high Cu (500 microg/g)/adequate Se (0.2 microg/g); high Cu (500 microg/g)/supplemented Se (2 microg/g); adequate Cu (18 microg/g)/adequate Se (0.2 microg/g); or, adequate Cu (18 microg/g)/supplemented Se (2 microg/g). Five rats per group were euthanatized after 3, 6, and 12 months, and liver specimens were obtained for histologic examination, histochemistry, metal analysis by atomic absorption spectrophotometry, measurement of glutathione peroxidase activity, and assessment of lipid peroxidation, using quantification of malondialdehyde (MDA) by the thiobarbituric acid reaction. RESULTS: Hepatic Cu concentration was significantly higher in rats fed high Cu diets (range, 9 to 18 microg/g of tissue [wet weight]), compared with rats receiving adequate Cu diets (4.0 to 5.7 microg/g of tissue). Rats fed high-Cu diets for 3, 6, and 12 months had mild multifocal hepatitis often surrounding necrotic foci. However, an increase in hepatic MDA content, indicative of lipid peroxidation, was not detected in these rats. Development of morphologic changes was not prevented by use of dietary Se supplementation. CONCLUSION AND CLINICAL RELEVANCE: Long-term exposure to excess dietary Cu caused mild hepatic lesions in Fischer 344 rats. Dietary Se supplementation did not prevent hepatic damage in rats with Cu toxicosis.     

亚慢性镉中毒对鸡体内NO代谢影响及硒颉颃效应的研究

亚慢性镉中毒可致鸡血清和肝脏组织匀浆中一氧化氮合酶(NOS)活性及一氧化氮(NO)含量升高,构成了亚慢性镉中毒损伤肝脏的病理学基础.补硒后提高机体的抗氧化能力.     

啤酒废酵母硒对肉鸡组织硒含量的影响

本试验旨在研究啤酒废酵母硒对肉鸡组织硒含量的影响。选用225羽1日龄平均体重为(42.78±1.08)g的AA商品代肉鸡(公母混合),随机分成5组,每组3个重复,每个重复15羽,对照组日粮为以Na2SeO3形式在基础日粮中补充硒0.1mg/kg,试验Ⅰ、Ⅱ、Ⅲ、Ⅳ组日粮为以酵母硒形式在基础日粮中补充硒0、0.1、0.2、0.3mg/kg,饲养期42d。结果表明:(1)肉鸡羽毛、肌肉和肝脏中硒的含量随着酵母硒添加量的增加和时间的延长显著提高,并表现出较强的线性关系;(2)与基础日粮组相比,酵母硒组肉鸡组织硒含量极显著增加(P0.05),以0.3mg/kg的添加量最好;硒的沉积顺序为肝脏羽毛肌肉。本试验结果提示,由啤酒废酵母制备的酵母硒可以提高肉鸡组织硒含量。     

Sequential morphological changes in chicken erythrocytes after in vivo and in vitro exposure to phenylhydrazine-hydrochloride

In acute haemolytic anaemia of chickens, induced with the oxidant chemical phenylhydrazine-hydrochloride, maximum degenerative changes in the in vivo exposed erythrocytes occurred on day 3 after injection. Microspherocytic transformation, dumb-bell shaped red blood cells and foamy squashed nuclei predominated. Heinz body formation was the cytoplasmic hallmark of the haemolytic anaemia. Marked reticulocytosis on day 5 indicated spontaneous regeneration. In vitro exposure to the chemical agent evoked similar morphological aberrations/Heinz bodies, microspherocytic transformation and nuclear degeneration. These were induced much earlier, presumably because of the absence of a protective internal milieu. Nevertheless, a basic similarity in morphology was evident. The in vitro test system might be suitable for screening oxidant chemicals and drugs.     

Quantitative changes of lung tissue components during perinatal period in rats

Quantitative changes of lung tissue components (air spaces lined by PAS-positive and PAS-negative epithelium, blood vessels and interstitium) were investigated in developing rats from fetal day 18 through neonatal day 1. The volume of the left lung increased significantly from fetal day 18 through neonatal day 1. The percentage and volume of the air spaces increased strikingly between fetal days 20 and 21. However, the percentage of the air spaces lined by PAS-positive epithelium decreased significantly from fetal days 20 to 21, and that of the spaces lined by PAS-negative epithelium increased between the two days. The proliferating cell nuclear antigen (PCNA)-positive cells were rich in the interstitium and epithelium of the air spaces on fetal days 18 and 19. The percentage of the interstitium decreased significantly from fetal day 18 through neonatal day 1, showing remarkable decrease between fetal days 20 and 21. From fetal day 20 onward, the PCNA-positive cells decreased in number and located in the epithelium of the conducting air ways and interstitium. Based upon these findings, the present study suggests that the period from fetal days 20 to 21 is a critical time for the development of fetal lung: the period before fetal day 20 is that for proliferation and the period after fetal day 21, functional differentiation of the lung.     

Effects of a single intraperitoneal administration of cadmium on femoral bone structure in male rats

Background

Exposure to cadmium (Cd) is considered a risk factor for various bone diseases in humans and experimental animals. This study investigated the acute effects of Cd on femoral bone structure of adult male rats after a single intraperitoneal administration.

Methods

Ten 4-month-old male Wistar rats were injected intraperitoneally with a single dose of 2 mg CdCl₂/kg body weight and killed 36 h after the Cd had been injected. Ten 4-month-old males served as a control group. Differences in body weight, femoral weight, femoral length and histological structure of the femur were evaluated between the two groups of rats. The unpaired Student''s t-test was used for establishment of statistical significance.

Results

A single intraperitoneal administration of Cd had no significant effect on the body weight, femoral weight or femoral length. On the other hand, histological changes were significant. Rats exposed to Cd had significantly higher values of area, perimeter, maximum and minimum diameters of the primary osteons'' vascular canals and Haversian canals. In contrast, a significant decrease in all variables of the secondary osteons was observed in these rats.

Conclusions

The results indicate that, as expected, a single intraperitoneal administration of 2 mg CdCl₂/kg body weight had no impact on macroscopic structure of rat''s femora; however, it affected the size of vascular canals of primary osteons, Haversian canals, and secondary osteons.     

孕鼠补硒对新生乳鼠十二指肠抗氧化能力的影响

为探讨孕鼠补硒对新生乳鼠十二指肠抗氧化能力的影响,将9只健康Wistar孕鼠随机分为低剂量补硒组(0.4mg/L,n=3)、高剂量补硒组(0.6mg/L,n=3)和对照组(0.0mg/L,n=3)。母鼠分娩后取新生乳鼠,测体质量,计算新生乳鼠的脾脏和胸腺指数,并测定十二指肠内谷胱甘肽过氧化物酶(GSH-Px)、总超氧化物歧化酶(TSOD)的活性和总抗氧化能力(T-AOC)。结果显示,与对照组相比,试验组新生乳鼠的初生体质量没有显著变化(P0.05),胸腺指数和脾脏指数均显著升高(P0.01),十二指肠内GSH-Px、T-SOD和T-AOC均显著降低(P0.01)。结果表明,孕鼠补硒降低了新生乳鼠十二指肠的抗氧化能力。     

Electrophoretic examination of changes in the protein spectrum of dogs after exposure to beta, beta'-dichlorodiethylsulfide]

In serum proteins, the alpha 3 + 4-globulin subfraction increases with a corresponding decrease of albumins. A statistically significant rise was observed from the 24th-48th hour from application with a progressive tendency reaching its maximum in the terminal stage of intoxication. This elevation reaches the values of 100% of the initial control values in the terminal stage. The drop in albumins is less significant and almost reciprocal with the rise of alpha-globulin content. In the values of beta-globulins, a change was observed in the quotient of beta 1/beta 2 globulins. No changes were ascertained in the values of gamma-globulins. Our results correspond to the data published by Gjessing and Chanutin, who observed a drop in albumins and rise in alpha-globulins after percutaneous implantation of the substance. The results suggest that the latent time between implantation and a statistically significant rise of the alpha-globulin fraction in the serum is 24-48 hours.     

Microbiologic studies of the lung clearance of rats after aerogenous long term exposure to ammonia

The effects of aerogenic long-time exposure of 1,990 female Wistar rats to 30, 50, 100 and 150 nl/l of NH3 for 3, 14, 28 and 42 days on clearance of inhaled bacteria (Staphylococcus aureus, SG 511 strain) were studied together with effects on the clinical condition and behaviour of these test animals. For concentrations below 100 nl/l, action upon pulmonary clearance was found to depend above all on the length of exposure, whereas effects of 150 nl/l proved to depend more strongly on concentration. All concentrations tested tended to act on pulmonary clearance in the form of increased numbers of bacteria which remained in the lungs of the experimental animals. Clinical symptoms attributable to NH3 action were not recordable.     

Anaemia and changes in erythrocyte morphology associated with copper and selenium deficiencies and dietary restriction in rats

Both selenium and copper deficiencies in rats caused morphological changes in erythrocytes but only copper deficiency resulted in anaemia. Similar but less severe morphological changes occurred when rats consuming selenium- and copper-adequate diets had their food intake restricted to that of the copper-deficient animals. Since selenium deficiency did not affect the severity of changes due to copper deficiency this suggests that alterations of erythrocyte morphology may not be entirely due to impaired functions of selenium and copper in cell antioxidant systems. Thus, erythrocyte structure was sensitive to several dietary changes and such alterations are probably independent of the anaemia induced by copper deficiency. This possibility should be considered when changes in erythrocyte morphology are attributed to copper or selenium deficiency in animals which also have a restricted food intake.     

Cardiovascular changes after bone marrow transplantation in dogs with mucopolysaccharidosis I

Five dogs with mucopolysaccharidosis I, 3 of which had been treated with bone marrow transplantation (BMT), were evaluated for 20 months with electrocardiography, thoracic radiography, and M-mode and 2-dimensional echocardiography. Treated and untreated (control) dogs had widened P waves on ECG. Thoracic radiographs remained normal for all dogs throughout the study. Thickening of the mitral valve was observed on echocardiograms of dogs in both groups, but the untreated dogs appeared to have thicker valves. Concentrations of glycosaminoglycans in the mitral valves and myocardium were higher in control dogs than in treated dogs. Markedly large aortic root diameters were observed on echocardiograms in both untreated dogs, but aortic root diameters remained normal in treated dogs. Echocardiography, but not electrocardiography, was useful in monitoring heart enlargement in each dog. Dogs treated with BMT generally had less severe cardiac changes and slower disease progression than control dogs.     

The effects of soybean selenium proteinate on tissue selenium and meat quality traits in finishing pigs

The purpose of the present study was to evaluate the effects of soybean selenium proteinate on Se tissue retention and meat quality in pigs. In group A (n = 11) the mixtures were supplemented with soybean selenium proteinate, in group B (n = 11) with sodium selenite and in group C (n = 11) with Se-enriched yeast (0.3 mg Se per kg in all groups). The use of soybean selenium proteinate resulted in lower retention of Se in tissues (liver, heart, muscle) compared to Se-enriched yeast. Selenium concentrations in tissues achieved by soybean selenium proteinate and sodium selenite were comparable. No differences in serum Se, serum GSH-Px and meat quality traits were found among the groups.     

The interaction of cadmium and selenium in horse kidney cortex in relation to histopathological changes

The kidney cortex of 32 Finnish horses was analysed chemically for cadmium (Cd) and selenium (Se) content and by light microscopy for histopathological changes of the tissues. Cd concentrations in kidney cortex ranged from 6.9 to 91.6 mg/kg wet weight with an average of 31.9 mg/kg. Se concentrations ranged from 0.5 to 1.5 mg/kg with an average of 1.0 mg/kg. The age of the horses varied from 1.5 to 32 years; mean age was 16 years.Cd levels in kidney cortex seemed to increase linearly up to an age of about 16 years. In old (over 16 years) horses no such correlation could be found. Se concentrations were found to decrease in relation to age. The horses with higher than 1 mg/kg Se in kidney cortex had less PAS-positive casts than horses with low Se content. Among the total population of the horses no correlation could be demonstrated between Cd and Se concentrations in kidney cortex. However, the horses with more than 40 mg/kg Cd in their kidney cortex had a slightly positive correlation between Cd and Se concentrations.