姜黄素对过度训练致大鼠脾脏细胞凋亡的调控作用及其机制*

目的:研究姜黄素调控Keap1-Nrf2-ARE信号通路缓解大鼠过度训练所致脾脏氧化应激及细胞凋亡机制。方法:7周龄SPF级雄性Wistar大鼠分为对照组(C组,n=12)、过度训练组(OM组,n=11)、姜黄素+过度训练组(COM组,n=14)。C组不进行任何运动干预,OM组、COM组大鼠进行8周递增负荷游泳训练。训练期间,COM组以200 mg/(kg·d)、5 ml/kg姜黄素进行灌胃,其他组灌胃等体积0.5 %羧甲基纤维素纳助溶剂。末次训练后24 h,称重计算脾脏指数,光镜观察脾脏组织病理学改变,取血液、脾脏组织检测相关生化指标。结果:C组大鼠脾脏组织结构正常;OM组较C组脾脏指数极显著降低(P＜0.01),并出现明显病理学改变;COM组较OM组脾脏指数显著升高(P＜0.05),且组织形态学改变有所改善。与C组比较,OM组血清皮质酮(Cor)浓度和脾脏细胞凋亡水平、丙二醛(MDA)浓度均升高,促凋亡蛋白Bcl-2相关X蛋白(Bax)表达增强(P＜0.05或P＜0.01);体重、血清睾酮(T)水平及脾脏超氧化物歧化酶(SOD)活性降低,脾脏血红素氧合酶1(HO-1)、抗凋亡蛋白B淋巴细胞瘤因子-2(Bcl-2)表达减弱(P＜0.05或P＜0.01);脾脏核因子E2相关因子2(Nrf2)表达水平无显著变化(P＞0.05)。与OM组比较,COM组体重无显著变化(P＞0.05);血清T浓度升高,脾脏SOD活性升高,Bcl-2、Nrf2和HO-1表达增强(P＜0.05或P＜0.01);血清Cor浓度及脾脏MDA浓度、细胞凋亡水平、Bax表达均降低或减弱(P＜0.05或P＜0.01);组间T/Cor比值变化趋势与T变化相一致,Bcl-2/Bax比值变化趋势与Bcl-2变化相一致。结论:8周递增负荷过度游泳训练引发脾脏细胞凋亡加剧,脾脏组织发生病理改变及功能异常。姜黄素通过上调Nrf2、HO-1蛋白表达,在一定程度上缓解过度训练引发的氧化应激,增强抑凋亡蛋白Bcl-2表达,减弱促凋亡蛋白Bax表达,改善大鼠脾脏细胞过度凋亡,保护脾脏组织结构和功能正常。     

The effect of internal exposure to 90Sr on hematopoietic stem cells in CBA mice

After acute intake of 90Sr the changes of d-9 CFUs number in mice (CBA) bone marrow, spleen and peripheral blood were investigated. The obtained results indicated similar quantitative changes in bone marrow and spleen CFUs on exposure to the 90Sr when radiation doses did not cause the decrease in life-time (1.11 kBq/g). Sarcomogeneous doses of 90Sr (29.6 kBq/g) resulted in drastic changes of hemopoietic system: spleen haematopoiesis activation and suppression of bone marrow functions. On the first day after 90Sr injection (29.6 kBq/g) the increase in number of peripheral blood CFUs (circulating pool) was observed.     

次声暴露对大鼠脾、肝脏某些酶活性的影响

目的:观察8 Hz,130 dB次声暴露不同时间对大鼠脾、肝脏某些酶活性的影响.方法:35只SD大鼠随机分为5组,即对照组,1周,2周,3周,4周组.每天次声暴露1次,每次2 h.实验后,观察大鼠脾、肝脏组织中MAO,GSH-px,SOD活性和MDA含量的变化.结果:大鼠脾脏MAO活性1周,2周时显著增高(P＜0.01),3周下降,4周时又显著增加(P＜0.05).肝脏组织MAO活性变化不明显(P＞0.05).脾脏组织中GSH-px活性在4周时明显增高(P＜0.05),肝脏组织中GSH-px活性在1周时就有显著性增高(P＜0.05).脾脏SOD活性在1周至4周均有显著性增高(P＜0.05).肝脏组织在实验期变化不明显(P＞0.05).脾脏组织中MDA含量在3周至4周时有显著性增高(P＜0.05).肝脏组织在1至2周时有非常显著的增高(P＜0.01),在3周时下降,到4周时又显著高于对照组(P＜0.05).结论:8Hz,130 dB次声暴露,大鼠脾、肝脏组织活性氧自由基、脂质过氧化物增高,抗氧化能力降低,造成对组织的损伤.     

Age-related growth of the spleen in normal and glucocorticoid treated rats

1. Age-related changes in the growth, nucleic acid content and protein turnover of the spleen have been studied in normal male rats. 2. A rapid and marked atrophy of the spleen, was found 24 hr after exposure to cortisone or dexamethasone; increased rates of protein breakdown being primarily responsible. 3. Nonetheless, the total amount of protein synthesized in the spleen (measured in vivo) was significantly decreased (30-50%) 24 hr after exposure to these steroids. 4. This compared with only a 15% decrease in whole body protein synthesis, indicating a more pronounced hormonal effect on the spleen than on most other body tissues.     

Spleen Volume Variation in Patients with Locally Advanced Non-Small Cell Lung Cancer Receiving Platinum-Based Chemo-Radiotherapy

There is renewed interest in the immune regulatory role of the spleen in oncology. To date, very few studies have examined macroscopic variations of splenic volume in the setting of cancer, prior to or during therapy, especially in humans. Changes in splenic volume may be associated with changes in splenic function. The purpose of this study was to investigate variations in spleen volume in NSCLC patients during chemo-radiotherapy. Sixty patients with stage I-IIIB NSCLC underwent radiotherapy (60Gy/30 fractions) for six weeks with concomitant carboplatin/paclitaxel (Ca/P; n = 32) or cisplatin/etoposide (Ci/E; n = 28). A baseline PET/CT scan was performed within 2 weeks prior to treatment and during Weeks 2 and 4 of chemo-radiotherapy. Spleen volume was measured by contouring all CT slices. Significant macroscopic changes in splenic volume occurred early after the commencement of treatment. A significant decrease in spleen volume was observed for 66% of Ca/P and 79% of Ci/E patients between baseline and Week 2. Spleen volume was decreased by 14.2% for Ca/P (p<0.001) and 19.3% for Ci/E (p<0.001) patients. By Week 4, spleen volume was still significantly decreased for Ca/P patients compared to baseline, while for Ci/E patients, spleen volume returned to above baseline levels. This is the first report demonstrating macroscopic changes in the spleen in NSCLC patients undergoing radical chemo-radiotherapy that can be visualized by non-invasive imaging.     

Tissue-specific hypomethylation and expression of rat phosphoenolpyruvate carboxykinase gene induced by in vivo treatment of fetuses and neonates with 5-azacytidine

Rat fetuses of 17-19-day gestation were injected in utero with 5-azacytidine (two to three daily injections of 40 micrograms/fetus). Neonates were injected with seven daily injections (1 mg/kg). DNA samples were isolated from the fetal and neonatal livers and neonatal spleen and subjected to analysis of their methylation status. Overall methylation was analyzed by the nearest-neighbor analysis (at CpG sites) and the pattern of methylation at CCGG sites by Southern blot analysis using phosphoenolpyruvate carboxykinase (PEPCK) sequences as probes. While DNAs from the liver and spleen undergo hypomethylation to the same extent in response to the 5-azacytidine treatment, the changes in the methylation patterns of the PEPCK gene in the two tissues are strikingly different. The changes observed indicate that a decrease in the methylase activity (inhibition by 5-azacytidine) results in site- and tissue-specific hypomethylation. The tissue-specific changes in the methylation pattern are associated with a tissue-specific expression of the PEPCK gene. Although the gene is hypomethylated by azacytidine in both liver and spleen, it is expressed only in the liver. The expression of already active genes (PEPCK in the kidney and albumin in the liver) is not further enhanced by the drug.     

Dynamics of the endo-DNAase activity of cell nuclei of mouse thymus and spleen lymphocytes during the immune response

Endo-DNAse (mostly Ca/Mg-dependent endonuclease) activity was studied in extracts of lymphocyte cellular nuclei from the spleen and thymus of mice upon their immunization with sheep red blood cells. Endo-DNAses were detected by their action on super-stranded DNA pBR 322. It has been established that endo-DNAse activity considerably changes in the course of immune response. The changes start in the early (induction) phase of immune response, are characterized by certain regularities and are distinct in thymus and spleen lymphocytes. It is assumed that endo-DNAses of lymphocyte cellular nuclei are involved in antigen-dependent lymphocyte differentiation.     

A comparative study of the kinetic changes of hemopoietic stem cells in mice infected with lethal and non-lethal malaria.

The kinetic changes of hemopoietic stem cells in bone marrow and spleen were compared between lethal Plasmodium berghei- and non-lethal P. yoelii 17x-infected mice. P. yoelii 17x-infected mice showed more severe splenomegaly than those infected with P. berghei. P. yoelii 17x-infected mice also showed a greater degree of sustained increase in number of multipotent hemopoietic stem cells (colony-forming units in spleen: CFU-S) and committed stem cells for granulocytes and macrophages (CFU-GM) and for erythrocytes (CFU-E) than P. berghei-infected mice. Such an increase was predominantly seen in the spleen of P. yoelii 17x-infected mice. In P. berghei-infected mice, the number of CFU-S, CFU-GM and also CFU-E only transiently increased and then decreased to a subnormal level at the late stage of infection. The proportion of cycling CFU-S was higher in P. berghei-infected mice than in P. yoelii 17x-infected mice. The IL-3 producing activity per spleen was much higher in P. yoelii 17x-infected than in P. berghei-infected mice at any point in time during the infection. Thus, hemopoietic changes seen after malaria infection seem to be closely related to the pathogenicity of the malaria parasite.     

The role of antioxidative status in development of the consequences of biological action under low dose and low intensity irradiation

The scale and direction of changes of a number of biophysical, biochemical and physiological parameters in SHK mice (male) tissues were studied depending on their initial values for mice control groups within 1 day after low intensity gamma-irradiation (15 cGy). The reciprocal dependences between the scales of the lipid antioxidative activity (AOA) in brain or the spleen mass changes and their values for mice control groups were found. The external dependences were revealed between the amounts of lipid peroxidation secondary products in spleen, liver and blood plasma of the control mice and a scale of their change after irradiation. The most substantial changes of this parameter were observed in blood plasma and the changes of the phospholipid content within the total lipid composition were found in spleen and blood erythrocytes of the irradiated mice, that is in the lipids of tissues, which had the lowest level of AOA for mice control groups. The experimental data obtained indicate that the initial antioxidant status of animal tissues plays the important role for the development of consequences of the biological action under low dose and low intensity irradiation.     

Antioxidant activity of fluoxetine: Studies in mice melanoma model

In vivo effects of the antidepressant fluoxetine on spleen antioxidant status of C57BL/6 mice were studied using a melanoma experimental model. After a 14‐day treatment with fluoxetine (10 mg kg^?1 day^?1, i.p.), the endogenous antioxidant non‐enzyme (glutathione) and enzyme (superoxide dismutase (SOD) and glutathione peroxidase (GPx)) defense systems in spleen of healthy animals were not changed; the lipid peroxidation (LP) was also unchanged. When B16F10 melanoma cells were introduced in C57BL/6 mice 2 h before fluoxetine treatment, a drug‐protective effect against the melanoma‐induced oxidative changes (increased LP and decreased total glutathione (GSH)‐level, as well as antioxidant enzyme activities) in spleen was observed. Fluoxetine dose‐dependently reduced the amounts of free oxygen radicals (hydroxyl and superoxide anion radicals), generated in chemical systems. Taken together, the present results suggest that fluoxetine, acting as antioxidant, prevents from melanoma‐induced oxidative changes in mice spleen. Copyright © 2010 John Wiley & Sons, Ltd.     

Studies on the lymphoid system of mice with lethal acute toxoplasmosis.

Acute toxoplasmosis was induced in CFLP mice by infecting them intraperitoneally with the 25 x 10(3) multiplicity of the virulent RH strain of Toxoplasma gondii. The lymphoid system of mice succumbing to acute toxoplasmosis showed characteristic changes. Significant spleen hypertrophy (spleen index: 1.76), severe thymus atrophy (thymus index: 0.27) and a striking decrease of the lymphocyte count in blood (86%) was found as compared with the uninfected controls.     

Specificity,avidity, and size of B-lymphocyte populations during ontogeny

The size and specificity of plaque-forming cell precursors (PFC) in murine fetal liver, neonatal, and adult spleen were studied in an adoptive transfer system. In this system, anti-4-hydroxy-3-iodo-5-nitrophenylacetic acid (anti-NIP) and anti-2,4,6-trinitrobenzene sulfonic acid (anti-TNP) direct PFC are generated from bone marrow-derived (B) cell precursors in fetal liver between 17 and 20 days of gestation and in 6- or 14-day neonatal spleen. PFC generated from fetal liver and neonatal and adult spleen cells are specific in that they lyse either NIP-coupled SRBC or TNP-coupled SRBC but not both. The generation of specific anti-NIP and anti-TNP PFC from precursors in fetal liver is primarily independent of antigenic stimulation. In contrast, the anti-NIP and anti-TNP responses generated from neonatal and adult spleen are antigen dependent. Both high-avidity PFC (detected with SRBC indicators coupled at low hapten density) and low-avidity PFC (detected with SRBC coupled at high hapten density) are generated from fetal liver and neonatal spleen cells; however, the proportion of high-avidity PFC precursors in adult spleen is at least threefold greater than in fetal liver or neonatal spleen. Analysis by velocity sedimentation indicates that most high-avidity PFC precursors are small lymphocytes in fetal liver, medium lymphocytes in 6-day neonatal spleen, and small lymphocytes in 14-day-old and adult spleen. Low-avidity PFC precursors are primarily medium-sized lymphocytes in fetal liver and 6-day neonatal spleen. In 14-day-old and adult spleen almost all high- and low-avidity PFC precursors are small lymphocytes. The results are discussed in terms of relative changes in the pool sizes of these lymphocyte populations.     

姜黄素对过度训练大鼠脾脏炎症反应的调控作用及其机制

目的:研究姜黄素调控Toll-样受体4(TLR4)-p38丝裂原活化蛋白激酶(p38 MAPK)/核因子κB(NF-κB)信号通路缓解过度训练大鼠脾脏炎症反应的作用及其机制.方法:7周龄SPF级雄性Wistar大鼠分为安静对照组(C组,n=12)、过度训练模型组(OM组,n=11)、姜黄素+模型组(COM组,n=14)...     

Involvement of interferon in virus-induced lymphopenia

Intraperitoneal injection of vesicular stomatitis virus (VSV) into mice causes marked and rapid changes in leukocyte distribution. The virus induces an increase in peripheral blood (PB) granulocytes and an extensive decrease in the lymphocyte count which reaches a nadir of less than 10% of preinfection values, 12 hr after virus inoculation. In the lymph nodes and spleen extensive lymphocyte translocation and granulocyte infiltration are observed. Most changes abate 48 hr following virus inoculation. Injection of poly(rI):(rC) causes similar changes to those observed with VSV. The lymphocyte changes observed after injection of VSV or poly(rI):(rC) coincide with high levels of Interferon (IFN) in the serum. We have examined the effects of anti-IFN antibody on those changes and investigated whether they can be mimicked by injecting IFN. Our findings suggest that the IFN induced by VSV or poly(rI):(rC), rather than those agents themselves, causes the observed lymphopenia as well as some of the changes observed in the spleen. On the other hand, the effects of VSV on granulocyte localization do not appear to be mediated by IFN.     

Hypoxia induces free radical damage to rat erythrocytes and spleen: analysis of the fluorescent end-products of lipid peroxidation.

Several studies have shown that hypoxia induces alterations in the lipid membranes of many cell types. The mechanism of these changes might consist in membrane lipid peroxidation. Lipid peroxidation in erythrocytes and spleen is easily detected by measurement of the concentration of fluorescent end-products. Exposure of rats to hypoxia for various time periods induced formation of lipophilic fluorescent products both in erythrocytes and spleen. A new kind of fluorophore was found in chloroform extracts from erythrocytes with excitation maximum at 270 nm and emission maximum at 310 nm. Additionally, two minor fluorophores were observed, emitting at 360 nm and in the region of 415-440 nm. Only one type of fluorophore was detected in spleen, emitting at 445 nm after excitation at 315 nm. The concentration of fluorophores was dependent on the time of hypoxic exposure both in erythrocytes and spleen. In erythrocytes there was a decrease of the predominant fluorophore after 3 hours (54%, P < 0.05) and 21 days (54%, P < 0.05) of hypoxia in relation to normoxic controls, accompanied by changes in spectral patterns of tridimensional fluorescence spectra. There was also a significant increase in the concentration of fluorophore in spleen (to 164%, P < 0.05, after 3 h, and to 240%, P < 0.05, after 21 days). The fluorophores, both in erythrocytes and spleen, were resolved into several distinct fractions with HPLC. The presented results support the hypothesis of hypoxia-induced lipid peroxidation and create a basis for further characterization of the fluorescent products.     

Spleen lipids: effect of whole body gamma irradiation and radioprotective chemicals

Effect of whole body gamma irradiation (1200 r) on spleen lipid metabolism of male and female rats 24 hrs and 48 hrs after irradiation and the effect of radioprotective chemicals vis. AET, serotonin, their mixture and cystamine on radiation induced changes in spleen lipid metabolism has been studied. In male rats both 24 and 48 hrs after irradiation a significant increase in spleen total lipids, cholesterol, phospholipids, phosphatidylethanolamine and phosphatidylcholine was observed. Administration of AET before irradiation prevented the changes in spleen total lipids and cholesterol but not in phospholipids, which was prevented by prior administration of both serotonin and the mixture of serotonin and AET. In female rats 24 hrs after irradiation only spleen total lipids showed an increase which was prevented by prior administration of cystamine. In male rats, 24 hrs after irradiation the incorporation of NaH2 32PO4 (counts/min/ug PLP and counts/min/g spleen) into spleen total phospholipids, phosphatidylcholine and phosphatidylethanolamine was reduced and this was corrected by prior administration of AET. Serotonin and the mixture of serotonin + AET did not protect the specific activity of phosphatidy choline. In female rats irradiation increased the incorporation of NaH2 32PO4 into phosphatidylcholine, which was not prevented by prior administration of cystamine. The fatty acid composition of spleen lipid of female rats was profoundly altered 24 hrs after irradiation. Palmitic acid and oleic acid showed an increase whereas arachidonic and fatty acid above arachidonic acid showed an decrease, which were corrected by administration of cystamine before irradiation.     

Ligula intestinalis (L.) (Cestoda: Pseudophyllidea): plerocercoid-induced changes in the spleen and pronephros of roach, Rutilus rutilus (L.), and gudgeon, Gobio gobio (L.)

The effects of the plerocercoid of Ligula intestinalis (L.) (Cestoda: Pseudophyllidea) on the major lymphoid organs, the spleen and pronephros, of roach, Rutilus rutilus (L.), and gudgeon, Gobio gobio (L.), are described. The spleen of ligulosed roach showed a significant decrease in weight. Differential cell counts suggested this was due to a reduction in erythrocytes, despite significant increases in macrophages and vacuolated granulocytes. The spleen of gudgeon, which consisted almost entirely of erythrocytes, showed a slight reduction in weight in ligulosed fish. In both roach and gudgeon this decrease was independent of parasite burden. Differential cell counts of the pronephros from ligulosed roach revealed a significant decrease in neutrophils and increase in vacuolated granulocytes. In the pronephros of gudgeon, however, cell counts were unaffected by ligulosis. Ultrastructural observations included an apparent disintegration of vacuolated granulocytes and increased pinocytic activity in specialized endothelial cells in the spleens of ligulosed roach. Also, melano-macrophage centres and melano-macrophages increased in the spleen of ligulosed roach and gudgeon, respectively. The marked changes in spleen weight and differential cell counts in ligulosed roach and lack of such changes in ligulosed gudgeon correlate with the differential response to the parasite by these two fish species.     

Different coexpressions of arthritis-relevant genes between different body organs and different brain regions in the normal mouse population

Structural changes in different parts of the brain in rheumatoid arthritis (RA) patients have been reported. RA is not regarded as a brain disease. Body organs such as spleen and lung produce RA-relevant genes. We hypothesized that the structural changes in the brain are caused by changes of gene expression in body organs. Changes in different parts of the brain may be affected by altered gene expressions in different body organs. This study explored whether an association between gene expressions of an organ or a body part varies in different brain structures. By examining the association of the 10 most altered genes from a mouse model of spontaneous arthritis in a normal mouse population, we found two groups of gene expression patterns between five brain structures and spleen. The correlation patterns between the prefrontal cortex, nucleus accumbens, and spleen were similar, while the associations between the other three parts of the brain and spleen showed a different pattern. Among overall patterns of the associations between body organs and brain structures, spleen and lung had a similar pattern, and patterns for kidney and liver were similar. Analysis of the five additional known arthritis-relevant genes produced similar results. Analysis of 10 nonrelevant-arthritis genes did not result in a strong association of gene expression or clearly segregated patterns. Our data suggest that abnormal gene expressions in different diseased body organs may influence structural changes in different brain parts.     

Viral RNA kinetics is associated with changes in trace elements in target organs of Coxsackie virus B3 infection

Trace elements are pivotal for the host defense, as well as potentially important for viral replication and virulence. Studies of sequential changes in viral replication in target organs of infection are sparse and a possible association with changes in specific trace elements is unknown. In this study Balb/c mice were infected with Coxsackie virus B3 (CVB3). Results indicated that sequential changes in viral replication (RT-PCR) were related to changes in trace element (arsenic, copper, iron, selenium and zinc) concentrations (as determined by ICP-MS) on days 3, 5 and 7 of the infection in serum, heart, lung, liver, pancreas, kidney, spleen, intestine and brain. After an initial viral peak on day 3, viral load drastically decreased in all organs, i.e. by >99% (serum), 97% (lung), 98% (liver), 60% (pancreas), 95% (kidney) and 93% (spleen), except in the heart, intestine and brain in which viral load increased after day 3. Selenium decreased in all organs except the heart while arsenic decreased in all organs except the kidney, spleen and brain. Moreover, selenium was negatively correlated to viral load in serum, liver, pancreas and intestine. To conclude, these findings give evidence that trace elements are directly involved in the replication of CVB3.     

Ah receptor in spleen of rodent and primate species: detection by binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin

In many species systemic toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is manifested by a generalized wasting syndrome accompanied by a variety of specific organ changes including atrophy of the thymus and spleen. TCDD toxicity in most tissues is thought to be mediated by the Ah receptor. Although the spleen is a prime target for TCDD toxicity, the possible presence of Ah receptor in the spleen has not previously been investigated. Specific binding of [3H]TCDD to Ah receptor in spleen cytosols was assessed by velocity sedimentation on sucrose gradients. Ah receptor was detected in spleen cytosols from adult Rhesus monkeys (mean +/- SEM, 36 +/- 8 fmol/mg cytosol protein), fetal Rhesus monkeys (9 +/- 6), Sprague-Dawley rats (20 +/- 5), C57BL/6J mice (18 +/- 2), New Zealand white rabbits (19 +/- 2), and Hartley guinea pigs (15 +/- 2). Ah receptor was not detectable in spleen cytosol from genetically "nonresponsive" DBA/2J mice or from Golden Syrian hamsters, a species resistant to toxicity of TCDD. Molecular properties of Ah receptor from spleen were similar to those of the receptor from liver of the same species. The high Ah receptor content in spleen cytosols from those species that are most susceptible to TCDD toxicity is consistent with the view that the Ah receptor mediates TCDD toxicity in spleen as well as in other tissues.