Methionine-rich storage protein gene in the rice stem borer, Chilo suppressalis, is expressed during diapause in response to cold acclimation

Gene expressions of acclimatized and non-acclimatized diapausing larvae were examined in Chilo suppressalis using a subtraction technique. A gene encoding a methionine-rich storage protein, CsSP1, was cloned and its complete cDNA sequence was determined. Potentially, CsSP1 encoded a 758-amino acid protein, with a calculated molecular weight of 88.8 kDa. The expression level of CsSP1 was higher in nondiapausing larvae than in diapausing ones. The CsSP1 expression was up-regulated in diapausing larvae when the temperature of cold acclimation was shifted to 5 degrees C. The up-regulated level was maintained at 40 days after incubation at 5 degrees C. In nondiapausing larvae, CsSP1 expression was down-regulated when the temperature was below developmental zero. Involvement of CsSP1 in diapause, cold tolerance acquisition and postdiapause development in C. suppressalis is discussed.     

压力超负荷致小鼠心肌肥厚中miR378对热休克转录因子1的调节作用

目的:探讨miR-378在压力超负荷致小鼠心肌肥厚中对热休克转录因子1(HSF1)的调节作用及机制。方法:采用C57B/L6雄性小鼠经主动脉缩窄构建心肌的压力超负荷(TAC)模型,2周后对小鼠进行心脏超声和血流动力学测定,并观察miR-378和HSF1的表达变化。体外培养心肌细胞,机械牵张刺激24 h后观察miR-378和HSF1的表达变化;对心肌细胞分别转染miR-378模拟物和抑制物,48 h后观察心肌细胞内源性HSF1的表达变化;通过荧光素酶基因报告系统,检测miR-378是否靶向结合其预测靶点HSF1的3′UTR区域。结果:建模TAC小鼠2周后,心脏表现为代偿性心肌肥厚,HSF1表达升高而miR-378表达下降。心肌细胞机械牵张后HSF1表达升高而miR-378表达下降;心肌细胞过表达miR-378后,HSF1表达显著下降,而当抑制内源miR-378时,HSF1表达显著升高;荧光素酶报告系统证实miR-378能够靶向结合HSF1的3′UTR序列。结论:在心肌肥厚代偿期,miR-378可能通过靶向结合HSF1的3′UTR序列调节心肌内源性HSF1的表达。     

局部微波热疗对热休克蛋白表达的影响

目的探讨骨肉瘤术中热疗对热休克蛋白表达的影响。方法对61例骨肉瘤组织进行免疫组化染色,观察热休克蛋白HSP-60、HSP-70及GP-94的表达。结果骨肉瘤组织中GP-94表达最高,阳性率为100%,其次为HSP-60,阳性率为94.5%,HSP-70弱表达,阳性率41.8%。热疗后HSP均不同程度增多,以HSP-70较明显,阳性率上升到72%。体外骨肉瘤细胞株加热后热休克蛋白也明显增多。结论局部微波热疗后,3种热休克蛋白在骨肉瘤组织均有表达。     

Cloning and interspecific altered expression of heat shock protein genes in two leafminer species in response to thermal stress

Studies have demonstrated differences in temperature tolerance between two Liriomyza species, L. huidobrensis and L. sativae. To investigate whether the heat shock proteins (Hsps) in the two species have different expression profiles during temperature stress, we cloned hsp90, 70, 60, 40 and 20, and analysed their expression profiles across temperature gradients by real-time quantitative PCR and Western blotting. The results revealed that the number of TATA-box-like elements and A/T-rich insertion/deletions within the 5' UTRs of the hsps are different in the two species. The temperatures for onset (T(on)) or maximal (T(max)) induction of hsp expression in L. huidobrensis were generally 2.5-10 degrees C lower than those in L. sativae, and the T(on) were highly consistent with the temperature limits of the northern boundary of the range of these two leafminer species. These studies confirmed, in terms of gene expression levels, that L. huidobrensis is more cold tolerant than L. sativae, which is more heat tolerant, and suggest that the T(on) (or T(max)) of hsps can represent the differences in temperature tolerance of these two leafminer species, and may be used to determine their natural geographical distribution limits.     

运动应激状态下的心肌热休克蛋白表达

背景：热休克蛋白因其具有特殊的生物学特性,在运动应激下探讨此类蛋白质在心肌的表达情况具有重要的研究价值。目的：了解国内外有关运动应激状态下心肌热休克蛋白表达的研究情况,对不同运动应激下心肌热休克蛋白的表达特点及意义进行分析。方法：以＂heat shock protein;myocardium;exercise stress＂为英文检索词;以＂热休克蛋白;心肌;运动应激＂为中文检索词,检索1991年1月至2014年1月检索PubMed数据库及万方数据库与热休克蛋白的产生、分类及功能有关、与运动应激下心肌热休克蛋白的表达相关、与运动应激下心肌热休克蛋白表达的机制相关及运动应激下心肌热休克蛋白表达的意义相关的文献。最终选择48篇进行归纳总结。结果与结论：热休克蛋白具有免疫协同作用。研究报道,运动训练能引起心肌热休克蛋白的表达。急性运动应激能使机体发生许多的生理、生化变化,心肌热休克蛋白会做出相对应的表达,对心肌细胞进行保护。低强度运动能够诱导增加热休克蛋白72基因的表达,抑制心肌细胞凋亡。大强度运动减少了热休克蛋白72基因的表达,不能有效抑制心肌细胞凋亡,不利于心肌的保护作用。中等运动强度下,热休克蛋白在不同运动时间的表达情况存在异议。运动诱导热休克蛋白表达对运动所致损伤的防护是有可能的,适量的运动活动在增强心肌功能、预防心肌在各种应激情况下的损伤具有重要作用。     

Can hyperbaric oxygen be used as adjunctive heart failure therapy through the induction of endogenous heat shock proteins

Heart failure (HF) is a chronic condition that is expected to increase in incidence along with increased life expectancy and an aging population. As the incidence of HF increases, the cost to national healthcare budgets is expected to run into the billions. The costs of lost productivity and increased social reliance on state support must also be considered. Recently, acute myocardial infarction (AMI) has come to be seen as the major contributing factor to HF. Although thrombolysis may restore coronary perfusion after an AMI, it may also introduce ischemic reperfusion injury (IRI). In an attempt to ameliorate sustained protein damage caused by IRI, endogenous chaperone proteins known as heat shock proteins (HSPs) are induced as a consequence of the stress of IRI. Recently, hyperbaric oxygen has been shown to induce the production of HSPs in noncardiac tissue, with a resultant protective effect. This current opinion review article suggests a possible role for hyperbaric oxygen, as a technologically modern drug, in augmenting the induction of endogenous HSPs to repair and improve the function of failing hearts that have been damaged by AMI and IRI. In addition, this simple, safe, noninvasive drug may prove useful in easing the economic burden of HF on already overextended health resources.     

严重烧伤早期肠黏膜组织热休克蛋白70的表达规律

目的探讨大鼠烧伤后早期肠黏膜组织热休克蛋白70(HSP70)的表达变化规律及其意义。方法采用大鼠烫伤模型,通过逆转录-聚合酶链反应(RT-PCR)、蛋白质免疫印迹(Westernblot)及免疫组化等方法,检测伤后3、6、12、24和48h不同时间点肠黏膜组织内HSP70及热休克因子1(HSF1)的表达分布情况。结果烫伤后3h肠黏膜组织内HSP70mRNA及蛋白表达均显著增加,分别在伤后6h和12h达高峰,伤后48h仍高于正常对照组(P均<0.01);伤后3h大鼠肠黏膜组织HSF1出现一过性降低,伤后6h其表达显著高于正常对照组,并呈逐渐增加的趋势直至持续到伤后48h(P均<0.01)。结论严重烧伤早期肠黏膜组织HSP70及HSF1表达均显著增加,提示严重烧伤早期即可引起肠黏膜组织细胞的应激反应,可能与细胞的自我保护机制启动有关。     

cDNA cloning of heat shock proteins and their expression in the two phases of the migratory locust

The high population density of insects is often a stress factor. Insects synthesize heat shock proteins (Hsps) in response to the impacts of stress through molecular chaperone activity. Locust solitary and gregarious phases occur at low and high population density, respectively. In this study, we compare the expression profiles of the Hsp genes in the two phases of the migratory locust in an attempt to examine the role of Hsps in adaptation to high density populations. The full length cDNAs of Hsp20.5, 20.6, 20.7, 40, 70 and Hsp90 of the migratory locust were cloned and sequenced. The expressional differentiation of the six Hsps in mRNA levels between solitary and gregarious locusts was observed. Results from real-time PCR indicate that the six Hsps are expressed throughout all developmental stages except in the early stage embryo. The expression levels of the six Hsps were significantly upregulated in gregarious locusts. The expressional variations among certain organs, such as the head, thorax and leg of fifth instar nymphs in gregarious locusts were also higher than those in solitary ones. These observations suggest that population density may be an important factor in determining Hsp expression in the locust.     

Are heat shock proteins involved in autoimmunity?

Summary Heat shock proteins (HSPs) have been postulated to be critical antigens in both autoimmune disease and experimental models of autoimmunity. This postulate has been largely based on the remarkable conservation of aminoacid sequence between human and bacterial HSPs, so that it has been argued that immune responses initially directed against the HSP of an infectious agent, would have the potential to initiate or maintain autoimmune disease. This would apply especially to T cell recognition of HSPs, since the T cell focuses on short peptide epitopes within a protein antigen rather than on the antigen's secondary structure. This article critically evaluates the available experimental evidence relating to this hypothesis: although research has clearly highlighted the central role of HSPs in the cellular immune response to pathogenic organisms and has shown the potential for T cell responses directed against self HSPs, a role for self HSPs as major target antigens in autoimmune disease has yet to be firmly established.     

热休克蛋白与肿瘤细胞耐药

热休克蛋白(HSP)是在热应激和有害的理化因素作用下,细胞应激产生的一组高度保守性的蛋白质,具有广泛的生物学功能。近年来研究表明,HSP与肿瘤细胞耐药关系密切,已成为肿瘤耐药研究的一个新靶点。本文综述HSP参与肿瘤耐药的机制及克服对策。     

谷氨酰胺对COPD外周血单个核细胞HSP70、TNF-α表达的影响

目的探讨谷氨酰胺（Gin）对慢性阻塞性肺疾病患者外周血单个核细胞（PBMC）中热休克蛋白70（HSP70）和肿瘤坏死因-α（TNF-α）表达的影响。方法选择30例COPD急性加重期（AECOPD）患者为研究对象,设为AECOPD组．将治疗10～20d后处于稳定期的上述患者设为SCOPD组,分别以Gin干预,并以健康人为正常对照组。采用Real—timePCR法检测上述5组PBMC中HSP70mRNA和TNF-α mRNA的表达水平。结果AECOPD和SCOPD空白对照组HSP70mRNA、TNF-α mRNA的表达高于正常对照组,差异有统计学意义（t分别=-3．74、-3．57、-3．63、-3．31,P均〈0．05）;AECOPD组和SCOPD组中,用Gin干预的较未用Gln的PBMC中HSP70表达均升高,差异有统计学意义（t分别=4．87、2．81,P均〈0．05）,而TNF-α表达均下降（t分别=-3．42、-2．97,P均〈0．05）。结论Gin可抑制COPD患者炎症因子TNF-α的活性,升高HSP70的表达。     

热休克蛋白70对严重创伤休克后肝脏的作用

目的 探讨热休克蛋白70（HSP70）对严重创伤休克后肝脏作用.方法 成年Wistar大鼠,采用双侧股骨骨折伴失血性休克致严重创伤模型,动态观察伤后8 h大鼠肝组织HSP70、血清肝功能生化指标、肝脏病理等变化.HSP70表达测定采用免疫印迹法,并进行计算机图像分析.结果 伤后HSP70在肝组织中表达迅速增加,6 h达到峰值,伤后8 h仍维持较高水平;创伤合并休克后,HSP70表达高峰提前至伤后4 h,持续表达至伤后6 h后逐渐下降,死亡前在肝组织中仍有少量表达.创伤休克后血清ALT、TB伤后4 h开始明显增高（P＜0.01）,白蛋白下降（P＜0.01）.肝脏镜下创伤休克后6 h肝窦内出现较多炎性细胞浸润.结论 在创伤休克早期,HSP70可能参与了肝组织细胞抗损伤机制的启动,但随着休克时间的延长,HSP70的过高持续表达,则可能对肝脏造成损害.HSP70在创伤休克后肝保护与肝损害过程中可能发挥双重作用.     

Induction of heat shock protein 70 genes in human lymphocytes during fever therapy

The induction of heat shock protein 70 ( HSP70 ) genes has been studied in peripheral blood mononuclear cells of individuals undergoing fever therapy because of metastatic malignant melanoma. Induction of HSP70 was assessed at the protein level by metabolic labelling or, for the HLA-linked HSP70-1 and HSP70-2 genes, at the RNA level by in situ hybridization. However, de novo expression of HSP70 could be observed during fever (usually above 39°C) in only about half of the cases. No simple threshold model for inducibility of HSP70 in vivo could be applied. The HSP70-1 gene was induced more easily than HSP70-2 . Thus, heat-inducible HSP70 genes, including HLA-linked HSP70 genes, become expressed in human lymphocytes during fever, but not regularly.     

臂丛神经损伤对热休克蛋白和神经型一氧化氮合成酶表达的影响

目的:观察臂丛神经远侧不同损伤后脊髓前角运动神经元中热休克蛋白70(Hsp70)、热休克蛋白27(Hsp27),以及神经型一氧化氮合成酶(nNOS)的表达变化.探讨其与运动神经元存活和再生的相互关系.方法:选用18只健康成年雄性SD大鼠,随机分为3组:肌皮神经切断组、挫伤组和对照组.术后2个月进行Grooming实验,检测其患肢运动功能的恢复.后取脊髓C5节段标本.观察脊髓运动神经元中三种蛋白的表达情况.结果:神经切断组和挫伤组的运动功能恢复均较明显,4级以上的百分比挫伤组(86%)优于切断组(57%).与对照组相比,两实验组损伤侧脊髓运动神经元中三种蛋白的表达均显著上调.切断组较挫伤组表达较高水平的Hsp70(140.61 1.25,130.08±2.38)和Hsp27(143.86±1.32,138.93±1.58);而nNOS蛋白在挫伤组中有较高水平的表达(挫伤组:106.12±0.91;切断组:68.18±0.63).结论:肌皮神经切断和挫伤后脊髓运动神经元中三种蛋白表达有所不同,其与损伤后功能恢复直接相关.     

糖皮质激素受体和热休克蛋白70在大鼠创伤失血性休克肝组织中的变化及其意义

目的从组织细胞受体水平探讨糖皮质激素受体（GR）和热休克蛋白70（HSP70）在创伤失血性休克后肝脏中的变化及其作用。方法采用双侧股骨骨折伴失血性休克致严重创伤模型，动态观察伤后8h大鼠肝组织GR、HSP70、血清肝功能生化指标、肝脏病理等变化。采用蛋白质免疫印迹法（Western blot）测定肝组织GR、HSP70含量，并进行计算机图像分析。结果单纯股骨骨折伤后，肝组织GR含量于伤后1h即开始下降，6h降至最低，8h仍显著低于正常对照组；创伤合并休克后，GR下降更加明显。HSP70在单纯股骨骨折伤后迅速增加，6h达到峰值，8h仍持续在较高水平；创伤合并休克后，HSP70升高更加显著。单纯股骨骨折伤后，血清丙氨酸转氨酶（ALT）、总胆红素（TB）和白蛋白与正常对照组比较，差异均无显著性。但创伤合并休克后4h和2h血清ALT和TB分别开始明显增高（P均d〈0．01），白蛋白则均下降（P均〈0．01）。创伤合并休克后6h肝脏镜下肝窦内即出现较多炎性细胞浸润。结论GR不足在严重创伤失血性休克后肝脏继发性损害过程中起重要作用，HSP70可能参与了肝组织细胞抗损伤机制的启动。     

抗热休克蛋白72多克隆抗体血清的制备和检测

目的 探讨在大肠杆菌中表达人热休克蛋白72（hHSP72）与组氨酸（His）的融合蛋白并制备其抗体血清的方法。方法 将hHSP72片断插入His融合表达载体pPROEX-1^TM，重组载体酶切鉴定后，在大肠杆菌中经异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达获得His-hHSP72融合蛋白。采用纯化后的HSP72免疫新西兰白兔，制备抗血清；蛋白质免疫印迹法（Western blot）检测重组抗原的免疫活性。结果 重组质粒酶切鉴定结果表明，hHSP72基因已正确插入到pPROEX-1^TM中，经IPTG诱导后，表达出分子质量约为73ku的蛋白，获得了效价为1：100000的多克隆抗体。Western blot检测证明，所制备的多克隆抗体可以与hHSP72特异性结合，其效价高于市售的单克隆抗体。结论 用hHSP72片断在大肠杆菌中能成功表达，并能制备得到多克隆抗体；这种多克隆抗体是一种新的高特异性和高灵敏度的试剂。     

The diagnostic value of urine heat shock protein 70 and prostatic exosomal protein in chronic prostatitis

ObjectiveTo explore the diagnostic value of the levels of prostatic exosomal protein (PSEP) and heat shock protein 70 (HSP70) in the urine of patients with chronic prostatitis (CP).MethodUrine samples from 210 CP patients (70 cases of the USA National Institutes of Health Category II [NIH‐II], 70 NIH‐IIIa, and 70 NIH‐IIIb patients) and 70 control subjects were collected between May 2018 and February 2020. The levels of PSEP and HSP70 in urine were detected by enzyme‐linked immunosorbent assay. The differences in urine PSEP and HSP70 levels between the groups were analyzed, and receiver operating characteristic (ROC) curves were used to analyze the clinical value of PSEP and HSP70 in the diagnosis of CP.ResultsThe PSEP levels of CP patients were significantly higher than those of the control group (p < 0.001), but there was no difference in PSEP levels among CP subgroups. The level of HSP70 in the urine of the NIH‐II patients was significantly lower than the levels in the NIH‐IIIa and NIH‐IIIb subgroups and the control group, but there was no difference in HSP70 levels between the NIH‐IIIa and NIH‐IIIb subgroups and the control group. ROC curve analysis results showed that the area under the curve (AUC) of PSEP for the NIH‐II, NIH‐IIIa, and NIH‐IIIb patients was 0.751, 0.776, and 0.731, respectively. The AUC of HSP70 in NIH‐II patients was 0.784, and the AUC of combined detection of PSEP and HSP70 in NIH‐II patients was 0.858.ConclusionUrine PSEP can be used as a marker for the diagnosis of CP, but it cannot distinguish between the various types of CP, and HSP70 can be used as a diagnostic index for NIH‐II classification.