Effects of nicotine and Vitamin E on Carbonic anhydrase activity in some rat tissues In Vivo and In Vitro

Effects of nicotine, nicotine+vitamin E and nicotine+Hippophea rhamnoides L. extract (HRe-1) on muscle, heart, lungs, testicle, kidney, stomach, brain and liver carbonic anhydrase (CA; EC 4.2.1.1.) enzyme activities were investigated in vivo. Groups of rats were given nicotine (0.5?mg/kg/day, i.p.), nicotine+vitamin E (75?mg/kg/day, i.g.), nicotine+HRe-1 (250?mg/kg/day, i.g.) and a control group vehicle only. The results showed that nicotine inhibited the heart, lung, stomach and liver CA enzyme activities by ～80% (p?<?0.001), ～94% (p?<?0.001), ～47% (p?<?0.001) and ～81% (p?<?0.001) respectively, and activated muscle and kidney, but had no effects on the testicle and brain CA activities. Nicotine+vitamin E inhibited the heart and liver CA enzyme activities by ～50% (p?<?0.001), and ～50% (p?<?0.001), respectively, and nicotine+vitamin E activated the muscle CA activity. However, nicotine+vitamin E had no effect on lung, testicle, kidney, stomach and brain CA activities. Nicotine+HRe-1 inhibited the heart and stomach CA enzyme activities by ～51% (p?<?0.001), and ～32% (p?<?0.002), respectively, and activated the muscle and brain CA activities, but had no effects on the lung, testicle, kidney, and liver CA activities. In vitro CA inhibition results for similar experiments correlated well with the in vivo experimental results in lungs, testicles, kidney, stomach, brain and liver tissues.     

Analysis of genes expressed in head kidney of common carp Cyprinus carpio L treated with cortisol

We analyzed genes expressed from head kidney of common carp Cyprinus carpio L. treated with cortisol. The results of single-pass sequencing of expressed sequence tags (ESTs) from 188 clones (AU240288–AU240367, AU301120–AU301227) from kidney cDNA are presented. One-hundred-twenty-seven clones (67.6%) were completely unknown and are likely to represent newly described genes, whereas 61 clones (32.4%) were identified based on matches to sequences in the database. The putative genes contain several ribosomal proteins, cytochrome oxidase subunits. Immune related cDNA clones identified from kidney were immunoglobulin light chain (n=4), FK506/rapamycin-binding protein (FKBP), CXC chemokine receptor type 4, complement factor B/C2-A3, peptidylprolyl isomerase (cyclophilin; Cyp)-like1, cyclophilin S1, heat shock-70 kDa protein-4, stress-activated protein kinase-3 (n=2). FKBP and cyclophilin genes expressed in normal tissues (head kidney, spleen, liver, brain and heart). Expression of FKBP and cyclophilin genes were not detected in liver, brain and heart when treated with cortisol for 16 h.     

Activation of mitogen-activated protein kinases during natural freezing and thawing in the wood frog

The responses of mitogen-activated protein kinase (MAPK) family members, including ERK (extracellular signal-regulated kinase), JNK (c-Jun NH₂-terminal kinase), and p38, in the metabolic responses to whole animal freezing (up to 24 h frozen at –2.5°C) and thawing (up to 4 h at 5°C after a 12 h freeze) were examined in four organs (liver, kidney, heart, brain) of the freeze-tolerant wood frog Rana sylvatica. Levels of the active phosphorylated form of p38 increased within 20 min as an early response to freezing in liver and kidney but rose later (after 12 h) in heart. Both JNK and p38 were activated during thawing in liver, kidney and heart with temporally-distinct patterns in each organ. The only MAPK response to freeze/thaw in frog brain was a transient elevation of p38 after 90 min thawing. ERK activity did not respond to freeze/thaw in any organ. The levels of c-Fos increased during freezing in kidney and brain whereas c-Jun was unaffected by freeze/thaw. Organ-specific responses by MAPKs, particularly p38, suggest that these may have roles in regulating metabolic or gene expression responses that may be adaptive in dealing with freezing stress or metabolic recovery during thawing.     

An Investigation of the Route and Progression of Encephalitozoon cuniculi Infection in Adult Rabbits

Rabbits infected either orally or intratracheally with cell culture-grown Encephalitozoon cuniculi were monitored regularly for serum antibody levels and E. cuniculi in the urine. Their responses were compared with intravenously inoculated and uninoculated control rabbits. All rabbits receiving E. cuniculi developed serum antibodies, generally within 3 weeks, and excreted E. cuniculi by 6 weeks. In the acute stage of infection, the organs most affected were lung, kidney and liver; the brain and gut were unaffected. However, during chronic infection, the brain, kidney, and heart were the only organs found to be involved. Antibody levels were very high at this stage. Thus both the oral and tracheal routes may be normal routes of infection with E. cuniculi in adult rabbits.     

Within species support for the expensive tissue hypothesis: a negative association between brain size and visceral fat storage in females of the Pacific seaweed pipefish

The brain is one of the most energetically expensive organs in the vertebrate body. Consequently, the high cost of brain development and maintenance is predicted to constrain adaptive brain size evolution (the expensive tissue hypothesis, ETH). Here, we test the ETH in a teleost fish with predominant female mating competition (reversed sex roles) and male pregnancy, the pacific seaweed pipefish Syngnathus schlegeli. The relative size of the brain and other energetically expensive organs (kidney, liver, heart, gut, visceral fat, and ovary/testis) was compared among three groups: pregnant males, nonpregnant males and egg producing females. Brood size in pregnant males was unrelated to brain size or the size of any other organ, whereas positive relationships were found between ovary size, kidney size, and liver size in females. Moreover, we found that the size of energetically expensive organs (brain, heart, gut, kidney, and liver) as well as the amount of visceral fat did not differ between pregnant and nonpregnant males. However, we found marked differences in relative size of the expensive organs between sexes. Females had larger liver and kidney than males, whereas males stored more visceral fat than females. Furthermore, in females we found a negative correlation between brain size and the amount of visceral fat, whereas in males, a positive trend between brain size and both liver and heart size was found. These results suggest that, while the majority of variation in the size of various expensive organs in this species likely reflects that individuals in good condition can afford to allocate resources to several organs, the cost of the expensive brain was visible in the visceral fat content of females, possibly due to the high costs associated with female egg production.     

Field and laboratory investigations of the thermal influence on tissue-specific Hsp70 levels in common carp (Cyprinus carpio)

Thermal discharge from power stations can affect normal environmental conditions and change in heat shock proteins expression of native fish with increasing temperature. In this study, we investigated levels of Hsp70 in the heart, kidney, brain and gill of the common carp Cyprinus carpio both in long-term heat discharge environment and after 24 h acute heat shock exposure. In laboratory exposure experiments, fish acclimated at 10 °C were exposed to various elevated temperatures (20, 24 and 28 °C). Hsp70 concentrations were determined in tissues by Western blotting analysis after one dimensional SDS-PAGE separation. In the field study, the level of Hsp70 in the gill of the carp remained at control values, and Hsp70 expression in the heart, kidney and brain underwent a 2.8 to 3.7-fold increase. A lower thermal sensitivity of the Hsp70 response of the brain, compared with the heart, kidney and gill, was observed in the laboratory experiments. Our data show that these tissues had different levels of Hsp70 responses to thermal influence both in acute exposure and long-term acclimation. The pattern of tissue Hsp70 expression may have a close relationship with the thermal tolerance of the carp and allows the fish to survive long-term thermal pollution.     

Effects of nicotine and vitamin E on glucose 6-phosphate dehydrogenase activity in some rat tissues in vivo and in vitro

Effects of nicotine, and nicotine + vitamin E on glucose 6-phosphate dehydrogenase (G-6PD) activity in rat muscle, heart, lungs, testicle, kidney, stomach, brain and liver were investigated in vivo and in vitro on partially purified homogenates. Supplementation period was 3 weeks (n = 8 rats per group): nicotine [0.5 mg/kg/day, intraperitoneal (ip)]; nicotine + vitamin E [75 mg/kg/day, intragastric (ig)]; and control group (receiving only vehicle). The results showed that nicotine (0.5 mg/kg, ip) inhibited G-6PD activity in the lungs, testicle, kidney, stomach and brain by 12.5% (p < 0.001), 48% (p < 0.001), 20.8% (p < 0.001), 13% (p < 0.001) and 23.35% (p < 0.001) respectively, and nicotine had no effects on the muscle, heart and liver G6PD activity. Also, nicotine + vitamin E inhibited G-6PD activity in the testicle, brain, and liver by 32.5% (p < 0.001), 21.5% (p < 0.001), and 16.5% (p < 0.001) respectively, and nicotine + vitamin E activated the muscle, and stomach G-6PD activity by 36% (p < 0.05), and 20% (p < 0.001) respectively. In addition, nicotine + vitamin E did not have any effects on the heart, lungs, and kidney G-6PD activity. In addition, in vitro studies were also carried out to elucidate the effects of nicotine and vitamin E on G-6PD activity, which correlated well with in vivo experimental results in lungs, testicles, kidney, stomach, brain and liver tissues. These results show that vitamin E administration generally restores the inactivation of G-6PD activity due to nicotine administration in various rat tissues in vivo, and also in vitro.     

Distribution of alpha1-adrenoceptor subtype proteins in different tissues of neonatal and adult rats

Distribution of alpha(1)-adrenoceptor (alpha(1)AR) subtype (alpha(1A), alpha(1B), alpha(1D)) proteins in brain, heart, kidney, and liver of 1-week-old rats and in brain, heart, aorta, kidney, liver, vas deferens, prostate, and adrenal glands of adult rats was investigated by Western analysis, using receptor subtype specific polyclonal antibodies. High levels of immunoreactive alpha(1A)AR and alpha(1D)AR in brain and heart and of alpha(1B)AR in liver and heart of neonatal rats were detected. In adult rat tissues, the abundance of alpha(1A)AR protein was most marked in the brain, intermediate in heart, aorta, liver, vas deferens, and adrenals, and minimal in the kidney and prostate; relative to other tissues, the expression of alpha(1B)AR was higher in brain and heart and that of alpha(1D)AR in brain. All the three receptor subtypes increased with age in the brain cortex, whereas the abundance of alpha(1B)AR increased in the heart but decreased in the liver; alpha(1A)AR and alpha(1D)AR in liver, kidney, and heart were not affected by age. It is concluded that alpha(1)AR subtypes are widely expressed in different neonatal and adult rat tissues.     

Carbohydrate contents, and glycosidase and glycosyl transferase activities in tissues from streptozotocin diabetic mice

The contents of hexoses and hexosamines in brain, liver, and kidney of streptozotocin diabetic mice are significantly increased in comparison to the controls. These differences for hexoses contents in the heart are not significant. N-acetyl-beta-D-glucosaminidase and beta-D-glucosidase activities in brain, liver and kidney of diabetic mice are significantly higher when compared to the controls. However, beta-D-galactosidase activity is significantly lower in brain, liver, spleen and kidney of the diabetic mice, in comparison to the controls and similar in heart. alpha-D-Mannosidase activity of diabetic mice is significantly increased in spleen and heart and significantly decreased in liver and kidney. alpha-L-Fucosidase of diabetic mice shows higher activities, with significant differences, in liver and spleen; however, in heart and kidney the activities are significantly lower. Brain sialyltransferase and galactosyltransferase activities are significantly increased in diabetic mice; but for heart and kidney these differences are not significant. The activity for brain and kidney fucosyltransferase is not significant and that for the other assayed organs is significantly higher in comparison to the controls.     

Effects of dibutyl phthalate and di‐ethylhexyl phthalate on acetylcholinesterase activity in bagrid catfish,Pseudobagrus fulvidraco (Richardson)

Acetylcholinesterase (AChE) activity is a well‐known biomarker for exposure to organophosphate or carbamate compounds in aquatic organisms. However, the effect of dibutyl phthalate (DBP) and di‐ethylhexyl phthalate (DEHP), widely used as a plasticizer, on the change of AChE activity is not yet known. Bagrid catfish Pseudobagrus fulvidraco were administrated with 100, 500 and 1000 mg kg^?1 diet of DBP or DEHP and the effects on AChE activity were assessed in the liver, gill, kidney, heart, brain, muscle and eye of the exposed fish. All tissues contained different background AChE activity in non‐treated bagrid catfish: the highest was observed in the brain, followed by muscle, heart, and kidney. The enzyme activities in various tissues were significantly inhibited after exposure to DBP or DEHP in a concentration‐dependent manner, especially in brain and muscle. A similar, but less pronounced, inhibition was also observed in liver and kidney when exposed to DBP and DEHP. Although AChE activity in gill and heart was also affected by DBP and DEHP, the decrease in these organs was least marked in these organs. Exposure to 1000 mg kg^?1 led to mortalities of 8.0% with DBH and 14% with DEHP; both seemed to be ascribable to phthalate toxicity. This study is the first report that the measurement of AChE activity in bagrid catfish is a valuable biomarker of DBP and DEHP exposure. This biomarker could be incorporated into a battery of biomarkers to strengthen the confidence with which ecotoxicologists can assess the impact of phthalate ester pollution in the aquatic environment.     

Alterations in antioxidant enzymes and oxidative damage in experimental diabetic rat tissues: Effect of vanadate and fenugreek (Trigonella foenum graecum)

With the premise that oxygen free radicals may be responsible for the severity and complications of diabetes, the level of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) as well as the oxidative damage were examined in the tissues of control, diabetic and treated rats. After three weeks of diabetes, the activity of CAT was significantly increased in heart in diabetes (about 6-fold) but decreased in liver. The SOD activity decreased significantly in liver but increased in brain. The activity of GPx decreased significantly in liver and increased in kidney. A significant increase was observed in oxidative damage in heart and kidney and a small increase in brain with decrease in liver and muscle. Vanadate and fenugreek (Trigonella foenum graecum) administration to diabetic animals showed a reversal of the disturbed antioxidant levels and peroxidative damage. Results suggest that oxidative stress play a key role in the complications of diabetes. Vanadate and fenugreek seeds showed an encouraging antioxidant property and can be valuable candidates in the treatment of the reversal of the complications of diabetes.     

Na,K-ATPase in several tissues of the rat: Tissue-specific expression of subunit mRNAs and enzyme activity

Summary The relative contents of Na,K-ATPase subunit mRNAs in rat renal cortex; ventricular myocardium, skeletal muscle (hind limb), liver and brain (cerebrum) were measured. Expressed per unit DNA, mRNA₁ content was 2-fold greater in the kidney and brain as compared to either heart, skeletal muscle or liver. The hierarchy of mRNA₂ expression was brain > skeletal muscle > heart, whereas mRNA₃ was restricted to brain. Betal subunit mRNA content in both kidney and brain exceeded the abundance of liver mRNA ₁ by 7-fold. In all tissues examined, the combined abundances of the alpha subunit mRNAs exceeded the content of mRNA ₁ The hierarchy of Na,K-ATPase activity expressed per unit. DNA was brain > kidney > skeletal muscle = heart > liver. The sum of mRNA as well as mRNA ₁ content, expressed per g of tissue, was highest in brain and kidney. A statistically significant correlation between mRNA ₁ content and Na,K-ATPase activity was evident.     

Fmr1基因敲除小鼠脏器重量和脏器系数的比较分析

目的通过对Fmr1基因敲除小鼠雌雄两性和FVB小鼠的脏器重量和脏器系数进行比较分析,了解其脏器重量的差异,探讨Fmr1基因对动物生长发育等方面的影响。方法分别测定Fmr1基因敲除小鼠雌雄两性和FVB小鼠内脏器官的绝对重量和脏器系数,并进行统计学处理和分析。结果相同年龄的Fmr1基因敲除小鼠雄性的体重、心、肺、肝和肾的绝对重量均极显著的大于雌性（P〈0.01）。雌雄间脏器系数除肾脏（P〈0.05）和脑（P〈0.01）外,其余无显著差异。与FVB小鼠比较,Fmr1基因敲除小鼠心脏较轻（P〈0.01）,肾脏（P〈0.01）、体重和脑较重（P〈0.05）。脏器系数肾脏较大（P〈0.01）,心脏（P〈0.01）、脑和脾（P〈0.05）较小。结论Fmr1基因可影响动物的某些脏器重量和脏器系数。     

Association of selenium with tissue membranes of ovine and rat tissues

Rats and lambs were injected with⁷⁵Se-selenite and the microsomal fraction obtained by differential centrifugation from homogenates of muscle, heart, liver, brain, kidney, and testes. These fractions were subsequently centrifuged on sucrose density gradients from 25 to 45% and the distribution of selenium-75 between the labeled membranes of different densities monitored by scintillation counting. The majority of the selenium-75 was associated with membranes of similar density in muscle, heart, liver, and testes from both rats and lambs. The selenium-75 was distributed between many different density membranes in brain from both lambs and rats. The selenium-75 was associated predominantly with one density membrane from ovine kidney but with several density membranes from rat kidney.     

Effects of nicotine and vitamin E on carbonic anhydrase activity in some rat tissues in vivo and in vitro

Effects of nicotine, nicotine + vitamin E and nicotine + Hippophea rhamnoides L. extract (HRe-1) on muscle, heart, lungs, testicle, kidney, stomach, brain and liver carbonic anhydrase (CA; EC 4.2.1.1.) enzyme activities were investigated in vivo. Groups of rats were given nicotine (0.5 mg/kg/day, i.p.), nicotine + vitamin E (75 mg/kg/day, i.g.), nicotine + HRe-1 (250 mg/kg/day, i.g.) and a control group vehicle only. The results showed that nicotine inhibited the heart, lung, stomach and liver CA enzyme activities by approximately 80% (p < 0.001), approximately 94% (p < 0.001), approximately 47% (p < 0.001) and approximately 81% (p < 0.001) respectively, and activated muscle and kidney, but had no effects on the testicle and brain CA activities. Nicotine + vitamin E inhibited the heart and liver CA enzyme activities by approximately 50% (p < 0.001), and approximately 50% (p < 0.001), respectively, and nicotine + vitamin E activated the muscle CA activity. However, nicotine + vitamin E had no effect on lung, testicle, kidney, stomach and brain CA activities. Nicotine + HRe-1 inhibited the heart and stomach CA enzyme activities by approximately 51% (p < 0.001), and approximately 32% (p < 0.002), respectively, and activated the muscle and brain CA activities, but had no effects on the lung, testicle, kidney, and liver CA activities. In vitro CA inhibition results for similar experiments correlated well with the in vivo experimental results in lungs, testicles, kidney, stomach, brain and liver tissues.     

Expression of orphan receptors GPR22 and GPR162 in streptozotocin-induced diabetic rats

Abstract

Aims/Introduction: Diabetes mellitus is a chronic degenerative disease characterized by high blood glucose levels as a result of problems in the action or insulin secretion. Although there are many treatments for this pathology, it has been associated with a high mortality rate. For this reason, it is important to try to identify new pathways that could be involved in diabetic complications. Recently, a new class of receptors has been reported, called orphan receptors because the associated ligand and signaling pathways are unknown, these receptors have been associated with certain pathologies. Therefore, the aim of this work was to study the expression of the orphan receptors GPR22 and GPR162 in heart, aorta, brain and kidney of diabetic rats. Materials and methods: We used Wistar male rats with 10–12 weeks of age. Diabetes was induced by a single dose of streptozotocin (60?mg/kg i.p.). After four weeks, the tissue was obtained and the expression of the mRNA was measured by RT-PCR. Results: Our results showed that the orphan receptors are expressed in a different way in heart, kidney, brain and aorta of diabetic and non-diabetic rats. Conclusions: We conclude that orphan receptors could be involved in the development of diabetes complications.     

小鼠组织中过氧化氢酶的活性与年龄的关系

为观察小鼠组织中过氧化氢酶的活性与年龄的关系,采用高锰酸钾滴定法测定不同年龄(1、4、18月龄)小鼠肝、肾、肺、心、脾、胃、脑组织中过氧化氢酶的活性。结果显示:小鼠过氧化氢酶在不同组织中活性不同,活性高低顺序基本表现为:肝>肾>肺>心、脾、胃>脑;小鼠肺、心、脾、胃、脑各组织中过氧化氢酶的活性在1～4月龄间随年龄增加而增加,在4～18月龄间随年龄增加而降低;小鼠肝、肾组织中过氧化氢酶的活性在1～4月龄间与年龄相关性不显著,在4～18月龄间随年龄增加而降低。结果表明,小鼠肝、肾、肺、心、脾、胃、脑等组织中过氧化氢酶的活性随年龄变化而变化,机体过氧化氢酶活性的降低与机体衰老密切相关。     

Immobilization stress in rat tissues: alterations in protein oxidation, lipid peroxidation and antioxidant defense system

We determined the effects of immobilization stress on antioxidant status, protein oxidation and lipid peroxidation in brain, liver, kidney, heart and stomach of rats. Sixteen male Wistar rats (3 months old) were divided into controls (C) and immobilization stress group (IS). IS rats were immobilized for 180 min/day for 15 days. Plasma corticosterone levels were increased in IS group. Copper,zinc-superoxide dismutase activities were increased in brain, liver and kidney, but decreased in the heart and stomach after immobilization. Catalase activities were increased in brain, kidney and heart, and decreased in liver and stomach. Selenium-dependent glutathione peroxidase activities were decreased in brain and kidney, but increased in heart and stomach. Reduced glutathione levels were decreased, while protein carbonyl, conjugated dienes and thiobarbituric acid-reactive substances levels were increased in all tissues. Our results showed that the response of antioxidant defense system to stress differs for each tissue, and protein oxidation and lipid peroxidation is induced by immobilization stress in peripheral tissues.     

Possible Role of Catalase in Adaptation to Diving of Semi-Aquatic Rodents Ondatra zibethica

Physiological role of catalase in mechanisms of adaptive reactions to hypoxic influences in diving rodents, muskrats Ondatra zibethica, has been investigated. The myocardial tissues were established to be the most resistant to the oxidative stress produced by hypobaric hypoxia-reoxygenation. An intraperitoneal injection of a specific nonreversible catalase inhibitor, 3-amino-1, 2, 4-triazol (3-AT), at a dose of 1 g/kg body weight, on the background of the diving hypoxia-reoxygenation decreased the catalase activity in liver, kidney, heart, and brain tissues, on average, by 83%. No statistically significant changes of this activity were observed in the femoral muscle tissue. In 56.5 min after the injection, the tissue concentrations of 3-AT in liver, kidney, heart, and femoral muscle were approximately similar to amount to about 1 µg/mg wet tissue. The 3-AT concentration in the brain was three times lower than in other tissues; however, this practically did not affect the extent of catalase inhibition. During the diving hypoxia-reoxygenation the reduction of catalase activity under effect of 3-AT was accompanied by an increase of lipid peroxidation (LPO) only in the heart and liver, which indicates an important role of catalase in protection of these tissues from damaging effects of active oxygen forms at resumed external respiration after the long diving.