热休克蛋白（HSPs）在胚胎发育中作用的研究进展

在胚胎发育,特别是早期胚胎发育时期,基因转录活跃、蛋白质大量合成,细胞的增殖和分化非常剧烈,细胞的环境在不断变化,细胞对外界刺激十分敏感。这个时期的HSPs变化和作用表现得非常突出。本文简要总结了近十年有关热休克蛋白在胚胎发育中作用的研究成果。根据胚胎发育地HSPs的依赖性、HSPs基因表达的发育阶段特异性和组织特异性、干扰胚胎发育中HSPs表达程序以后导致胚胎异常发育等现象推测：（1）热休克基因与和／功调控体形形成、肌肉及神经分化,而在胚胎发育中具有看家基因的功能;（2）热休克蛋白作为伴侣分子,通过介导新合成的和／或可逆变性的蛋白质正确折叠、装配、转动及促进不需要的和／或不可逆变性的蛋白质降解,参与胚胎的正常发育和保护胚胎不受不良刺激的影响。这方面的深入研究,必将有助于阐明胚胎发育、细胞增殖、细胞分化和去分化、细胞转化、生物适应性等的分子机理。     

大鼠肝生长发育过程中ACP、AKP、HSC70/HSP68和PCNA的变化(英文)

磷酸酶（ＡＣＰ、ＡＫＰ）在生物的机能分化中起重要作用,热休克蛋白（ＨＳＰｓ）是近几年发现的一类在胚胎发育、细胞生存中起重要作用的分子,无论是胚胎发育还是细胞结构和功能构建都和细胞增殖密切相关,增殖细胞核抗原（ＰＣＮＡ）是检测细胞增殖的良好指标。 本实验用组织化学、免疫组织化学、Ｗｅｓｔｅｒｎ印迹、酶的原位复性电泳、体视学分析等方法定性和定量分析了酸性磷酸酶（ＡＣＰ）（Ｆｉｇ．１＆２）、碱性磷酸酶（ＡＫＰ）（Ｆｉｇ．４＆５）、构成性热休克蛋白 ７０／诱导性热休克蛋白 ６８（ＨＳＣ７０／ＨＳＰ６８）（Ｆｉｇ．６）和ＰＣＮＡ（Ｆｉｇ．７＆８, Ｔａｂｌｅ１）在大鼠肝生长发育（从１４天胚胎到成体）过程中的动态变化。结果表明：（１）在大鼠肝生长发育过程中,ＡＣＰ有两个活性高峰期,其时段处于大鼠吃奶和吃饲料起始期（Ｆｉｇ．１＆２）;（２）在ＡＣＰ的第一个活性高峰期时,ＡＫＰ活性降低;而在ＡＣＰ的第二个活性高峰期时,正值ＡＫＰ的活性高峰期（Ｆｉｇ．３）;（３）ＡＣＰ活性高峰期也是ＰＣＮＡ含量高峰期;（４）ＨＳＣ７０／ＨＳＰ６８在刚断奶的幼鼠肝和成体肝中表达量较多,其他时段表达极少。根据上述结果推测：ＡＣＰ和ＰＣＮＡ通过调节细     

关于脑缺血的分子生物学研究

脑缺血的主要直接后果是脑缺氧。脑缺血或脑缺氧除引起一系列神经化学变化与蛋白质的合成降低外,还引起热休克蛋白（ＨＳＰ）和ｃ－ｆｏｓ蛋白等特殊蛋白质的特殊变化。轻度缺血可引起ＨＳＰ７０基因转录与翻译;中度缺血只引起其转录而不翻译;重度缺血时转录与翻译均终止。轻、中度脑缺血引起ｃ－ｆｏｓｍＲＮＡ剧烈而短暂的表达,ｃ－ｆｏｓ、ＪｕｎＢ、ｃ－ｊｕｎ等基因转录增加;严重缺血可能因神经元死伤,导致ｃ－ｆｏｓ蛋白水平降低,但局部胶质细胞ｃ－ｆｏｓ诱导增强。     

热休克蛋白——细胞的内源性保护蛋白

关于应激反应的最初报道是 1 962年Ｒｉ ｔｏｓｓａ等在果蝇唾液腺染色体中发现一种热诱导的“蓬松”现象 ,并具有基因转录活性[1] 。 1 974年Ｔｉｓｓｉｅｒｓ等从热休克果蝇唾液腺细胞中分离出一组特殊蛋白质即热休克蛋白 (ｈｅａｔｓｈｏｃｋｐｒｏｔｅｉｎ ,ＨＳＰ)。目前根据分子量ＨＳＰ可分为 6个家族 ,1 0 0～ 1 1 0ｋＤ大分子ＨＳＰ家族、ＨＳＰ90家族、ＨＳＰ70家族、ＨＳＰ60家族、ＨＳＰ40家族及 1 8～ 3 0ｋＤ的小分子ＨＳＰ家族。其中ＨＳＰ70是最保守和研究最为深入的一种[2 ] 。在正常情况下 ,ＨＳＰ70通过辅助新生肽的折叠…     

HSP70抗原呈递分子的研究进展

热休克蛋白７０（ＨＳＰ７０）是广泛存在于原核和真核细胞中的一组高度保守的蛋白质分子家族,其主要功能是促进与维持新生多肽的正确折叠,并且调节细胞的生长、发育、分化、死亡。ＨＳＰ７０还是免疫系统识别的主要抗原,在抗原的加工、处理和呈递以及效应Ｔ细胞的激活过程中扮演着重要角色。来自同源肿瘤成份的ＨＳＰ治疗小鼠肿瘤具有明显的效果,是肿瘤肽的高效传递系统,是ＣＤ＾＋８Ｔ细胞激活的佐剂。了解其功能对某些疾病发     

菜豆下胚轴质膜微囊和液泡膜微囊体外热稳定性的研究

从抗热潜势较高的“８５ＣＴ－４９７６２”菜豆（Ｐｈａｓｅｏｌｕｓ ｖｕｌｇａｒｉｓ Ｌ．）的下胚轴分离出微粒体、质膜微囊和液泡膜微囊。通过比较正常细胞的膜微囊、热锻炼细胞的膜微囊以及热锻炼过程中放线菌酮抑制了蛋白质合成的细胞膜微囊的质子泵在体外的热稳定性,发现热锻炼可使细胞膜微囊质子泵体外热稳定性提高,热锻炼过程中合成的蛋白质与此效应有关。进一步分析膜外周蛋白对质子泵体外热稳定性的影响,证明热激蛋白ＨＳＰ７０ 和小分子量热激蛋白（ＬＭＷ ＨＳＰ）对质子泵有热保护功能     

幽门螺杆菌热休克蛋白A基因的克隆、表达及免疫原性研究

幽门螺杆菌的感染可诱发人体产生胃炎和消化性溃疡,其组成成分热休克蛋白Ａ（ＨｓｐＡ）可刺激机体产生保护性的免疫反应。用ＰＣＲ方法从幽门螺杆菌的染色体ＤＮＡ上扩增出ＨｓｐＡ基因片段,将其插入原核表达载体ｐＥＴ２２ｂ（＋）中,并在ＢＬ２１（ＤＥ３）大肠杆菌表达。经测序ＨｓｐＡ基因片段有３５４ｂｐ组成,可编码１１８个氨基酸残基的多肽。ＳＤＳＰＡＧＥ和免疫印迹分析检测发现,ＨｓｐＡ基因表达的蛋白质分子量约为１５ｋＤ,并证实该重组蛋白质可以被幽门螺杆菌感染阳性患者的血清所识别,同时将其免疫小鼠可刺激机体产生抗该重组蛋白质的抗体。ＨｓｐＡ有可能作为一种有效的蛋白质疫苗用于幽门螺杆菌感染的预防和治疗。     

水稻热休克转录因子OsHSF13的克隆与生物信息学的初步分析(简报)

环境刺激的信号转导是植物信号转导的一个重要研究方向。热激反应(heat-shockresponse,HSR)是动植物细胞或器官在遇到外界热刺激时所产生的一种保护性反应,是正常的蛋白质合成受阻时产生热激蛋白(heat-shockprotein,HSP)的一种细胞生理活动,其表达通过热休克转录因子(heat-shock factor,HSF)来进行调控[1]。编码热激蛋白基因的启动子区域存在着一段保守的DNA序列,是热休克转录因子的结合位点(heat-shockelement,HSE)。当植物受到外界的热刺激时,HSF可以与HSE特异性结合,激活热激蛋白基因的表达,     

人工老化自理的卷心菜种子的热激蛋白合成

高活力卷心菜种子蛋白质合成速率比中等活力和低活力种子高很多。热激处理（４２℃）下,蛋白质合成显著下降,但高活力种子的蛋白质合成能力仍然显著高于中等和低活力种子。高活力和中等活力种子主要合成分子量为７０ｋＤ和一些小分子量的热激蛋白。在低活力种子中检测不到热激蛋白的合成。４种热激蛋白（１种ＨＳＰ９０和３种ＨＳＰ７０）的Ｗｅｓｔｅｒｎ ｂｌｏｔ检测结果表明,只有１种热激蛋白（ＨＳＰ７０）与种子活力有关。     

热休克蛋白的生物医学功能

热休克蛋白（Ｈｓｐ）是真核细胞和原核细胞在应激条件下产生的一种蛋白质,在正常状态的细胞中也广泛存在。Ｈｓｐ是生物种系发育中最保守的蛋白质之一,其主要功能是促进新生多肽的正确折叠并维持折叠状态,从而发挥调节细胞生长、分化、生存的功能。Ｈｓｐ还有调节免疫功能、参与疾病发生的作用。     

Circadian rhythmic changes in the lipid composition of muscles in kilka <Emphasis Type="Italic">Clupeonella cultriventris</Emphasis> (Clupeidae,clupeiformes) during natural periodicity of feeding in the feeding and spawning periods in the Rybinsk Reservoir

According to data of observations in 2002–2004, differences in the pattern of diurnal fluctuations in the total content and fractional composition of lipids in muscles of mature specimens of kilka Clupeonella cultriventris from the Rybinsk Reservoir in the feeding and spawning periods were revealed. It was established that the feeding intensity in kilka considerably changes throughout 24 h in both periods of the annual cycle, while diurnal fluctuations in the fatness of muscles are distinctly manifested only in the reproductive period and have a dissimilar pattern in specimens of different sexes. In females and males, they are determined mainly by the change in the content of the fraction of reserve lipids-triacylglycerols, as a rule, by its increase in the light hours of the day, several hours after an increase in the feeding activity of fish. The pattern of diurnal fluctuations of the level of lipid fractions (phospholipids, triacylglycerols, cholesterol, and its ethers) in kilka’s muscles differs from that in the feeding period. Possible causes of the change of diurnal variations of the considered indices of lipid metabolism in kilka at an increase in the endocrine activity of its body in the reproductive period is discussed.     

果实膨大期喷布磷酸二氢钾对三月红荔枝果实贮藏性的影响

在三月红荔枝(Litchi chinensis cv.Sanyuehong)果实膨大期对树冠喷施0.2%磷酸二氢钾(KP)水溶液,以探讨磷酸二氢钾对荔枝果实贮藏性的影响。结果表明:(1) KP处理的果实在贮藏期前17d,失重率及果肉可溶性固形物、酸、VitC、花色素苷等指标的变化趋势与对照基本相似;(2) KP处理的果肉可溶性蛋白质含量变化与对照有明显差异,而果皮的可溶性蛋白质含量在贮藏期前10d变化动态与对照一致,此后呈相反的变化趋势;(3)果皮POD活性显著高于果肉,KP处理和对照的果肉POD活性在贮藏第3d后、果皮POD活性在第17d前分别具有相似变化趋势;(4)果肉中CAT活性在贮藏第3~17d期间都显著或极显著高于果皮,KP处理和对照果肉、果皮CAT活性动态变化均为单峰曲线。     

Influence of in vitro factors on titre and elimination of model fruit tree viruses

The titre of apple chlorotic leaf spot virus (ACLSV) was higher in microplants of Malus domestica cv. Jonagold than in 2-year-old grafted scions. Cytokinin concentration in the medium increased the titre of prunus necrotic ringspot virus (PNRSV) in the apex of microplants of Prunus insititia cv. Kozlienka but did not affect the titre of ACLSV in M .domestica.Virus titre of ACLSVwas higher in the haulms of autotrophically-grown compared with heterotrophically-grown microplants whereas as for PNRSV the results were the reverse. For both viruses, however, titre of the virus in the roots of autotrophically-grown plants was significantly higher than in haulm tissue from heterotrophic cultures. Ribavirin incorporation resulted in elimination of both viruses. Negative ELISA results were confirmed independently by PCR. The efficacy of Ribavirin in elimination of ACLSV was increased by increasing the concentration of cytokinin in the medium in parallel with decreasing the concentration of Ribavirin. These results are discussed in the context of the reliability of in vitro virus testing.     

Rat brain thioltransferase: regional distribution, immunological characterization, and localization by fluorescent in situ hybridization

Thioltransferase (TTase) is a member of the family of thiol-disulfide oxidoreductases that are involved in the maintenance of sulfhydryl homeostasis in cells by catalyzing thiol-disulfide interchange reactions. One of the major consequences of oxidative stress in brain is the formation of protein-glutathione mixed disulfides (through oxidation of protein thiols), which can be reversed by TTase during the recovery of brain from oxidative stress. We therefore examined the presence of TTase in brain regions from rat. In the rat, TTase activity in the whole brain was comparable with the corresponding activity in liver, but significantly higher in hippocampus. The enzyme activity was significantly lower in striatum and cerebellum compared with activity in whole brain. Rat brain TTase shared immunological similarity with the human red blood cell enzyme, but not with the pig liver enzyme. The constitutive expression of the mRNA to TTase was demonstrable by northern blotting. Localization of the TTase mRNA in rat brain by fluorescent in situ hybridization showed the presence of high amounts of mRNA in the olfactory bulb, cortex, and hippocampus and its predominant localization in the neurons. TTase mRNA was also present in Purkinje cells in the cerebellum, in giant reticular neurons in the midbrain, and in the striatal and thalamic neurons. This study demonstrates the constitutive presence of a functional TTase system in brain and delineates the regional and cellular localization of the enzyme in rat brain.     

The effect of periovulatory imbalance of prolactin on the expression of prolactin receptors in rat ovary cells

Using the technique of immunohistochemistry in combination with cytophotometry, we have studied the effect of periovulatory hyper- and hypoprolactinemia on the expression of prolactin receptors in various cell types of rat ovaries during early estrus. It has been shown that intense specific staining of oocytes is positively controlled by prolactin. The maximal intensity of specific staining was found in cells of the cumulus and the inner layer of granulosa cells in mature follicles; staining intensity gradually diminished towards the outer boundary cell layer. Postovulatory follicles are distinct from those mature follicles in which there was no ovulation in their more intense manifestation of prolactin receptors in cells of the inner layer and cumulus, as well as in increased positive staining (after prolactin administration) only in the granulosa layer cells closest to theca. In follicles which did not ovulate by the time of the early estrus, prolactin administration leads to a proportional growth of specific immunoreactivity in all cell layers of the granulosa. The administration of bromocryptin, an inhibitor of prolactin secretion, leading to a 10-fold decrease in the prolactin level in the blood, results in a twofold decrease in the intensity of specific staining of all cell layers of the granulosa in either type of follicle. Corpora lutea of the previous cycle have irregularly positioned luteocytes with weak and strong specific staining, the intensity of which is not changed in response to prolactin and diminishes slightly after the administration of bromocryptin. We conclude that the most intense changes in the content of prolactin receptors under the conditions of imbalance of this hormone during the periovulatory period are observed in those follicles where the oocyte did not ovulate by the time of early estrus.     

Effect of Moniliformis moniliformis density on distribution within the definitive host population (Rattus norvegicus)

The population dynamics of Moniliformis moniliformis was studied in ‘free-ranging’ laboratory rats, Rattus norvegicus, presented with different relative density levels of M. moniliformis in cockroaches, Periplaneta americana. Changes in selected population parameters of the negative binomial distribution were evaluated as indicators of changes in aggregation. A significant increase in the degree of aggregation of parasites occurred as a result of the increase in relative density of infective stages available to the rats. This increase in aggregation was due to the increase in over-dispersion that occurred in female rats only. The degree of aggregation in females was found to be significantly higher than that in males at both treatment levels. The best indicators of the degree of aggregation were found to be the ratio of the variance to the relative density and the ratio of the log-variance to log-relative density. Changes in k were not correlated with changes in over-dispersion or the relative density.     

Antiatherogenic effect of a low lysine: arginine ratio of protein involves alteration in the aortic glycosamihoglycans and glycoproteins

The effect of alteration of lysine: arginine ratio of the protein on the aortic glycosaminoglycans and glycoproteins was studied in rats fed cholesterol free and atherogenic diet. The concentration of total glycosaminoglycans and of individual fractions was significantly lower in the aorta in the case of diet with lysine: arginine ratio of 1.0, than the diet with a ratio of 2.0. Rats fed globulin fraction isolated from sesame seeds, which has a lysine: arginine ratio of 0.67 also showed significantly lower concentration of total and individual glycosaminoglycan fractions in the aorta than those fed casein (lysine:arginine ratio 2.0). Concentration of total hexose and fucose in the glycoproteins was also lower in the aorta in the case of lysine: arginine ratio 1.0. These results in the light of previous reports of increase in the aortic glycosaminoglycans in the early stages of atherosclerosis and increase in the total hexose and fucose in the glycoproteins in the atherosclerotic aorta indicate that the antiatherogenic effect of a low lysine: arginine ratio in the protein involves alteration in the aortic glycosaminoglycans and glycoproteins.     

Modification of chemical properties of cell wall polysaccharides in the inner tissues by white light in relation to the decrease in tissue tension in Pisum sativum epicotyls

When white light irradiation inhibits shoot growth in derooted pea ( Pisum sativum L. cv. Alaska) cuttings, it decreases tissue tension, a driving force for shoot growth, by making the cell wall of the inner tissues mechanically rigid. To elucidate the mechanism by which light affects the mechanical properties of the cell wall in the inner tissues, its effect on the chemical properties of the cell walls was studied by analyzing qualitatively and quantitatively cell wall polysaccharides in the epdidermis and inner tissue of pea epicotyls grown in both dark and light. The amount of polysaccharides per subhook in the cell walls of both tissues increased during a 4-h dark incubation. Light suppressed the increase in hemicellulosic (HC)-II and cellulosic polysaccharides in the inner tissues, while it did not affect the increase in other wall fractions in either the epidermal or subepidermal tissues. No light effect was observed on the neutral sugar compositions of pectin, HC-I or HC-II fractions in either of the tissues. Light increased the mass-average molecular mass of xyloglucan and rhamnoarabinogalactan in HC-II fractions in the inner tissues, while such an effect was not observed in the epidermis. These facts suggest that the light-induced decrease in the tissue tension in pea epicotyls is caused by an increase in the molecular size of xyloglucan, rhamnoarabinogalactan in the HC-II fraction and/or the suppression of the synthesis of HC-II and cellulosic polysaccharides in the inner tissues.     

Ultrastructure and Lipase Activity of Cotyledon cell During Pod Development in Peanut

A series of significant changes of the ultrastructure and lipase activity of cotyledon cell were found in peanut (Arachis hypogaea) during pod development. In he initial stage of cotyledon development there were many plastids which kept producing starch grain and there were low lipase activity and very few lipid and protein bodies in the cell. In the middle stage of cotyledon development, a great number of larger lipid bodies were seen in the cell and a lot of protein bodies formed in the vacuoles and continued to increase in size. Lipase activity increased in the cytoplasm, endoplasmic reticulum, protein bodies, plasmalemma and intercellular space. In the later stage of cotyledon development, the lipid bodies did not increase in number but became slightly larger. The protein bodies continued to increase both in number and in size. Lipase acttvity was even hegher in the cytoplasm. In the final stage the protein bodies became irregular in shape and some of them tended to disintegrate with their content entered into the space around the lipid bodies. The lipase activity in the cell declined. The results indicated that the lipid body originated in the cytoplasm and the protein body originated in the vacuole; that the accumulation of oil and protein in peanut cotyledon resulted from the formation and development of lipid and protein bodies in the cell, and that the changes of plasmid and lipase activity in the cell played a role in the development of lipid body during the development of cotyledon.     

Relationship Between the Ubiquitin-Dependent Pathway and Apoptosis in Different Cells of the Central Nervous System: Effect of Thyroid Hormones

We have recently shown that sustained neonatal hyperthyroidism in the rat activates apoptosis of oligodendroglial cells (OLGc) and that inhibition of the proteasome-ubiquitin (Ub) pathway by lactacystin produces increased apoptosis in cerebellar granule cells (CGC). In the present study we have analyzed the relationship between the activation of the Ub-dependent pathway, the expression of the Ub genes and programmed cell death in neurons of the rat cerebellum and cerebral cortex and in OLGc. This study was carried out in normal animals, in rats submitted to sustained neonatal hyperthyroidism and in cell cultures treated with an excess of thyroid hormones. In neurons of the cerebral cortex, thyroid hormone produces an increase of Ub-protein conjugates, an enhancement in the expression of the Ub genes and an increase in apoptosis, while the opposite results are obtained in CGC. These results indicate that in neurons, the changes in the cell death program produced by thyroid hormone run in parallel with those occurring in the Ub-dependent pathway. In OLGc, thyroid hormone increases apoptosis but does not produce changes in the Ub pathway. Preliminary studies indicate that in coincidence with what occurs in optic nerves, the sciatic nerves both in controls and in hyperthyroid animals are unable to form Ub-protein conjugates. These results indicate that in cells of the CNS such as neurons, in which the Ub-dependent pathway is actively expressed, it appears to be closely correlated with apoptosis.