生长休止蛋白7在成年大鼠肾脏、心脏和肝脏中的差异表达

目的研究生长休止蛋白7（Gas7）在成年大鼠肾脏、心脏和肝脏的表达。方法成年SD大鼠16只,分别采用逆转录聚合酶链反应（RT-PCR）方法和免疫组织化学方法检测Gas7基因mRNA和蛋白在成年SD大鼠肾脏、心脏和肝脏的表达,并进行图像分析和统计学处理。结果RT—PCR结果显示,Gas7mRNA在肾脏高表达,在心脏的表达弱于肾脏（P〈0．05）,而在肝脏的表达最弱,基本检测不到。免疫组化结果显示,在肾脏中,Gas7免疫阳性产物在近髓肾单位的近曲小管呈强阳性反应,在集合管表达较弱,在肾小球和其余肾小管未见表达;在心脏中,Gas7免疫阳性产物均匀分布于心肌细胞,呈中等强度反应,弱于肾脏（P〈O．05）;在肝脏中,Gas7蛋白未见明显表达,与其mRNA在肝脏的表达相似。结论Gas7在大鼠肾脏、心脏和肝脏表达的不同,尤其在肾脏组织分布的差异性,提示Gas7在成年大鼠肾脏和心脏结构以及功能的维持中可能起着重要作用。     

Caffeic acid phenethyl ester ameliorates changes in IGFs secretion and gene expression in streptozotocin-induced diabetic rats

The protective effect of caffeic acid phenethyl ester (CAPE) against diabetes-induced alteration of IGFs protein and gene expression was investigated in serum, liver, heart, and kidney. In the present study, diabetic rats exhibited the decrease of IGF-I content in serum, liver and heart but the increase of that in kidney and CAPE blocked them. Diabetic rats also manifested the increase of IGF-II content in serum, liver, heart, and kidney and CAPE prevented them. CAPE prevented the diabetes-induced decrease of liver IGF-I mRNA and IGF-II mRNA, which is similar to pattern of IGFs mRNA in kidney. Moreover, diabetic rats exhibited the decrease of heart IGF-I mRNA but the increase of IGF-II mRNA and CAPE blocked them. In conclusion, CAPE, in part, prevented diabetes-induced alteration of IGF-I and IGF-II protein and gene expression in liver, heart, and kidney in rats.     

Appropriate expression of the mouse H19 gene utilises three or more distinct enhancer regions spread over more than 130 kb

H19 and Igf2 are closely linked, reciprocally imprinted genes which lie on distal chromosome 7 in the mouse. Data suggests that common elements are used for expression and imprinting of both genes, and simple models have been proposed based on the presence of a single set of enhancers located downstream of H19. In this study we have investigated the H19 expression pattern from a 130 kb YAC transgene, which imprints H19 appropriately at ectopic loci. However, we show that while enhancers for expression in many cell types are present on the YAC, those for expression in mesodermal components of the heart, kidney, lung and thymus are located at a greater distance. Based on the available evidence, we conclude that regulation of H19 is complex, requiring contribution from at least three different sets of cell-type specific enhancers. Thus, the mechanism of reciprocal imprinting of H19 and Igf2 utilises different regulatory elements in different cell types during mouse development.     

Comparisons between the inorganic content of healthy and hypertensive rat tissues by inductively coupled plasma-mass spectrometry

The inorganic contents of bone, brain, erythrocyte, heart, kidney cortex, kidney medulla, liver, lung, muscle and plasma from spontaneously hypertensive rats were compared with those of the same tissues from healthy Sprague-Dawley rats. A general inductively coupled plasma-mass spectrometry method developed for multi-element determinations of most of the elements present in biological tissues was used. Variations were found not only for major elements, as expected, but also for many trace elements in several tissues.     

Expression of muscle creatine kinase gene of mice and interaction of nuclear proteins with MEF-2, E boxes and A/T-rich elements during aging

The changes in the expression of muscle creatine kinase (MCK) gene in the heart and skeletal muscle of mice during aging were studied. Its expression declines as a function of age in the heart, however, no age-related change is observed in the skeletal muscle. The cis-acting elements, MEF-2, E boxes and A/T rich elements present in the enhancer region of the mouse MCK gene are known to regulate the expression of the gene. Hence, these elements were subcloned and electrophoretic mobility shift assay was carried out to investigate the changes in the binding of the nuclear trans-acting protein factors of the heart with these elements as a function of age. These factors showed specificity for the respective cis-acting elements. Furthermore, the binding of these factors was found to decrease during aging which may contribute to the age-related decline in the expression of the MCK gene and activity of the heart.     

Expression and developmental regulation of the NMDA receptor subunits in the kidney and cardiovascular system

Antagonists to the N-methyl-D-aspartate (NMDA) receptor bind to various extraneuronal tissues. We therefore assessed the expression of the main NMDA subunit, NR1, in various tissues. We demonstrate that NR1 appears to be most abundant in the rat kidney and heart. NR1 is present in total rat kidney, cortex, and medulla. Of the NR2 subunits, only the NR2C subunit protein is present in the kidney. The abundance of the NR1 subunit protein increases with kidney development. Both NR1 and NR2C are present in opossum kidney, Madin-Darby canine kidney, and LLC-PK(1) cells. Immunohistochemistry studies show that the NR1 subunit is present in the renal proximal tubule. NR1 is abundant in the atrium and ventricle but is also expressed in the aorta and pulmonary artery. The NR2 subunits are not expressed in the heart. NR1 subunit protein expression is constant throughout heart development. Finally, the NR1 subunit protein is expressed in heart cells (H9c2) grown in culture. These studies reveal the presence of the NMDA receptor in the kidney and the cardiovascular system.     

感觉神经损伤性盐敏感性高血压大鼠心、肾ACE和ACE2的表达

目的研究肾素-血管紧张素系统（RAS）基因血管紧张素转换酶（ACE）和血管紧张素转换酶2（ACE2）在感觉神经损伤性盐敏感性高血压大鼠心肌和肾脏中的表达情况,探讨ACE、ACE2在盐敏感性高血压发生发展中的作用。方法用乳鼠皮下注射辣椒辣素法建立模型。哺乳期后,大鼠被随机分成4组：对照＋正常盐饮食组（CON-NS）、对照＋高盐饮食组（CON-HS）、辣椒辣素＋正常盐饮食组（CAP-NS）、辣椒辣素＋高盐饮食组（CAP-HS）。四组大鼠分别接受4周不同的处理。至7周龄（分组饲养后第4周）处死大鼠,免疫组化检测大鼠心肌和肾脏ACE和ACE2蛋白的表达,反转录-聚合酶链式反应（RT-PCR）检测大鼠心肌和肾脏ACE和ACE2mRNA的表达。结果①至7周龄（分组饲养后第4周）各组动物体重差异无显著性（P〉0.05）。②各组动物在分组时（0周）鼠尾收缩压差异无显著性（P=0.583）,至7周龄（分组饲养后第4周）,CAP-HS组鼠尾收缩压明显高于其它三组（P〈0.01）。③心肌和肾脏ACE蛋白表达均升高。心肌组织,CAP-HS组与CON-NS比较,P〈0.01,与CON-HS和CAP-NS比较,P〈0.05;肾脏组织,CAP-HS组与其它三组比较,P〈0.01。④心肌和肾脏ACE2蛋白表达均降低。心肌和肾脏组织,CAP-HS组与CON-NS和CAP-NS比较,P〈0.01,与CON-HS比较,P〈0.05。⑤心肌和肾脏ACE mRNA表达均升高。心肌组织,CAP-HS组与CON-NS比较,P〈0.01,与CON-HS和CAP-NS比较,P〈0.05;肾脏组织,CAP-HS组与其它三组比较,P〈0.01。⑥心肌和肾脏ACE2 mRNA表达均降低。心肌和肾脏组织,CAP-HS组与CON-NS和CAP-NS比较,P〈0.01,与CON-HS比较,P〈0.05。结论感觉神经损伤性盐敏感性高血压大鼠心、肾ACE表达升高的同时有ACE2表达的降低,ACE和ACE2表达水平的差异可能与盐敏感性高血压的形成有关。     

感觉神经损伤性盐敏感性高血压大鼠心、肾AT1R的表达

目的对感觉神经损伤性盐敏感性高血压大鼠的心肌、肾脏组织中的血管紧张素Ⅱ1型受体（AT1R）在mRNA和受体水平的表达进行检测,探讨AT1R与盐敏感性高血压的关系。方法用乳鼠皮下注射辣椒辣素法建立模型。哺乳期后,大鼠被随机分成4组：对照＋正常盐饮食组（CON-NS）;对照＋高盐饮食组（CON-HS）;辣椒辣素＋正常盐饮食组（CAP-NS）;辣椒辣素＋高盐饮食组（CAP-HS）。至7周龄（分组饲养后第4周）处死大鼠,免疫组织化学方法和反转录-聚合酶链式反应（RT-PCR）分别检测大鼠心肌和肾脏AT1R蛋白,以及AT1 R mRNA的表达。结果①Wistar大鼠在给予不同程度的感觉神经损伤和饲料干预后,各组大鼠尾部收缩压均有明显增加,最终CAP-HS组的尾收缩压显著高于其他三组（P〈0.01）。②免疫组织化学结果显示,CAP-HS组组织有显著的AT1R蛋白表达（P〈0.01）;CON-HS组肾脏、心肌组织中AT1R蛋白表达高于CON-NS组（P〈0.05）。③RT-PCR检测基因表达,与对照组CON-NS相比,实验组CAP-HS的AT1R mRNA表达显著升高（P〈0.01）;CON-HS组肾脏、心肌组织中AT1 R mRNA表达有显著性（P〈0.05）。结论感觉神经损伤性盐敏感性高血压大鼠心、肾AT1R表达升高,AT1R表达水平的差异可能与盐敏感性高血压的形成有关。     

Trace elements in different tissues in aging rats

BackgroundTrace elements (TE) in the human body provide a connecting link between the environment, lifestyle and biochemical modulation of homeodynamics. On the other hand, many non-essential (toxic) elements are linked to numerous diseases. Our study tried to identify differences in TE levels between healthy old and young Wistar rats in blood and the tissues of kidney, liver, heart, and testicles. Furthermore, we wanted to see if there were age-related differences in correlations between essential and/or non-essential (toxic) TE within and between mentioned tissues.MethodsWe used 28 healthy male Wistar rats which were divided into two age groups: young, aged 10 weeks (n = 15) and old, aged 36 months (n = 13). The animals were sacrificed under general anesthesia and the blood samples, and samples from the tissues of the heart, kidneys, testicles, and liver were used for the determination of TE content in them. Analysis of the 16 elements was performed by inductively coupled plasma mass spectrometry (ICP-MS).ResultsToxic elements in old rats (As, Hg, and Cd) were significantly higher in all of the tissues where the difference in levels of these elements was found. Tissues of the kidney and liver had the most correlations between TE in old and young rats, respectively. In both old and young rats, arsenic was the toxic element that had most of the correlations with other essential or non-essential elements. In old rats, most of the TE correlations were detected between the tissues of the kidney and heart (11 correlations), while in young rats most of the correlations were observed between the tissues of kidney and liver, and kidney and testicles (with 9 correlations both).ConclusionsOur study has found significant changes in levels of trace elements in all of the mentioned tissues, with kidney and testicles being the tissues with the most TE differences between the two aged groups. This and other similar studies should encourage other investigators to evaluate the mutual connections between TE and physiological, or the “unhealthy” aging. More studies with more tissues included, more biomarkers of the systemic function, and even molecular methods are needed to provide the answers to numerous questions relating to TE and aging.     

脑红蛋白在赛加羚羊主要内脏器官中的表达与定位

赛加羚羊(Saiga tatarica)属于我国一级重点保护野生动物,其原产地主要为高寒低氧地区,现存种群则主要栖息于中亚地区的荒漠及半荒漠草原上。脑红蛋白是一种存在于脊椎动物体内具有运输和储存血氧能力的球蛋白,在动物适应低氧过程中具有重要的生理功能。为了初步探究赛加羚羊对低氧环境的耐受性机制,运用免疫组织化学染色法与实时荧光定量PCR技术,对脑红蛋白及脑红蛋白基因(NGB)在赛加羚羊的心、肝、脾、肺、肾等5种主要内脏器官中的分布规律与表达情况进行了探究。免疫组织化学染色结果显示,脑红蛋白在赛加羚羊的心、肝、脾、肺、肾中均有分布,阳性表达主要分布在其心肌细胞、肝细胞、脾白髓区中的淋巴细胞、肺泡细胞以及肾小球内皮细胞。实时荧光定量PCR结果显示,脑红蛋白基因在赛加羚羊心、肝、脾、肺、肾中的表达量不同,脾的表达量最高,心的表达量次之,两者均显著高于肝、肺和肾(P < 0.05);其后依次为肝、肾、肺,其中,肝的表达量显著高于肾和肺(P < 0.05),肾和肺之间表达量差异不显著(P > 0.05),肺的表达量最低。上述研究表明,脑红蛋白在赛加羚羊的主要内脏器官中均有阳性表达,不同内脏器官中的表达量不同,这表明脑红蛋白可能参与了这些内脏器官的氧利用过程,具体机制有待进一步探讨。     

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.     

Distribution of 14 elements in various rat tissues following hypophysectomy,thyroparathyroidectomy, adrenalectomy,and castration

The tissue distribution of 14 elements was simulatneously determined in rats 28 d after hypophysectomy (HPY), thyroparathyroidectomy (TPTY), adrenalectomy (ADY), and castration (CTN). The elements Na, K, Ca, Mg, Fe, S, P, Rb, Sr, Mn, Cu, and Zn were investigated in whole blood, plasma, brain, liver, kidney, heart, skeletal muscle, and bone. Additionally Mo was determined in kidney and liver. The following results were obtained: 1) With regard to hormone deficiency: HPY induced the most noticeable, variations on all the elements tested owing probably to the direct and indirect effects of adenohypophyseal hormones. ADY led to the expected modification of Na and K but also to a Sr accumulation and a Rb depletion. TPTY induced a sharp decrease in plasma and tissues Ca, an increase in plasma P, but did not disturb the two elements in bone. An increase of Rb in many tissues and of Fe in heart, kidney, and liver were also observed. CTN had little consequences except in bone whose Cu and Fe contents were increased: 2) With regard to element variations: K, Mg, and S underwent little change. Discriminations were revealed between elements such as K and Rb, Ca and Sr, Ca and Mg, and Cu and Zn. The changes of Rb and Sr were consistent with regulatory mechanisms. The accumulation of Fe and Cu in tissues such as liver after HPY, TPTY, and ADY, suggest that the hormonal deficiencies could worsen the hemochromatosis with Wilson's disease; 3), With regard to plasma and tissues: No correlation appeared in element levels between plasma and other tissues. Brain was the least affected and liver, kidney and bone the most.     

茶多酚干预乳腺癌组织及重要器官血管生成相关因子表达研究

目的:观察茶多酚对移植性小鼠乳腺癌(EMT_6)组织与重要脏器(心、脑、肾)组织血管生成相关因子表达影响。方法:应用小鼠可移植性乳腺癌EMT_6细胞株,经培养传代后,以纯系BALB/c小鼠为荷瘤动物进行移植,并采用茶多酚灌胃及局部注射两种干预措施,以免疫组化方法检测小鼠乳腺癌组织VEGF、bFGF、TIMP-2表达,并测定心、脑、肾组织中VEGF及TIMP-2表达。结果:与模型对照组比较,茶多酚两种给药途径的肿瘤组织VEGF、bFGF阳性表达明显降低(P<0.05);TIMP-2阳性表达明显增高(P<0.05);而心、脑、肾组织VEGF、TMP-2阳性表达无明显差异(P>0.05)。结论:茶多酚可明显抑制新生血管生成相关因子表达,并特异性的作用于肿瘤靶点部位,预示在肿瘤治疗领域具有广泛的应用前景。