The mRNA Expression Profiles of Five Heat Shock Protein Genes from Frankliniella occidentalis at Different Stages and Their Responses to Temperatures and Insecticides

The westem flower thrips, Frankliniella occidental＆ （Pergande） is a highly invasive pest that is able to exploit many crops across a wide range of environmental conditions. Five full-length cDNAs of heat shock protein （HSP） genes （Fo-HSP90, Fo-HSP70, Fo-HSP60, Fo-HSP40 and Fo-HSP28.9） were cloned from F. occidentalis, and their expression profiles were investigated under conditions of thermal stress and insecticide exposure, and at different stages during development, using real-time quantitative PCR. All five gene sequences showed high similarity to homologs in other species, indicating the conserved fimction of this gene family. HSP60 represents an informative phylogenetic marker at the ordinal taxonomic level within Insecta, but HSP90, which has two homologous copies in Hymenoptera, was not informative. The expression of Fo-HSPs under thermal stress suggests that Fo-HSP90, Fo-HSP70, and Fo-HSP28.9 are inducible by both cold and heat stress, Fo-HSP40 is only heat-inducible, and Fo-HSP60 is thermally insensitive. There were two patterns of cold induction of Fo-HSPs： one is from 0 to 4℃ and the other is around -8℃. All five Fo-HSPs genes were induced by exposure to sublethal concentrations of the insecticide avermectin. The expression of the five Fo-HSPs during different developmental stages suggests that they all play a role in development of F. occidentalis.

Identifying candidate genes and patterns of heat-stress response in rice using a genome-wide association study and transcriptome analyses

Because high temperatures impair rice production,it is desirable to elucidate the regulatory mechanisms involved in rice response to heat stress.The objectives of this study were to identify candidate genes and characterize their response patterns during rice adaptation to high temperatures at the seedling stage.Ten heat-associated quantitative-trait loci were identified in a genome-wide association study.Comparison of transcript abundances in heat-sensitive and heat-tolerant rice pools under heat stress revealed approximately 400 differentially expressed genes.The expression of genes from heatsensitive accessions changed more than those from heat-tolerant accessions under heat stress.Alternative splicing(AS)events responded to heat stress in rice.The types of AS variants significant different between the heat-sensitive and heat-tolerant accessions.Expression patterns differing between the heat-sensitive and heat-tolerant accessions were identified for genes known to be involved in heat stress.We identified eleven genes associated with rice heat stress response.These genes could be pyramided to breed heat-tolerant rice accessions.

Mining Heat Stress Associated Genes in Tomato Fruit (Solanum lycopersicum L.) Through RNA-seq

Tomato(Solanum lycopersicum L.)is a thermophilic vegetable crop,but sensitive to high temperature stress,especially under the greenhouse conditions.Due to global climate changes,heat stress has now become a great threat to tomato production and fruit quality.Many studies have been conducted to determine the functions of genes in tomato responsive to abiotic and biotic stresses,but transcriptomic information on heat stress responses of tomato fruit is still limited.To investigate heat stress associated genes in tomato fruit,a cDNA library was constructed using fruit harvested from tomato cv.P19-9 plants grown under 42℃for 0,1,2 and4 h and the expression profiles of heat stress responsive genes in tomato fruit were analyzed through RNA-seq.A total of 632224558 clean high quality paired-end reads were obtained and then mapped to reference genome for RNA-seq analysis.After quality control analysis,alignment analysis and transcript assembly,a total of 55457 RNA transcripts were obtained with functional annotations.Overall,6869 differentially expressed genes(DEGs)were identified with a significant response to one or more of the three heat stress treatment times.Based on GO enrichment analysis,22 genes potentially involved in tomato thermo-tolerance were selected and validated for their expressions through qPCR.The expression profile of tomato fruit genes obtained in this study could shed light on the mechanism and gene editing breeding projects for tomato thermo-tolerance.These findings could also benefit improvement of harvest and storage of tomato in greenhouse.

Microarray-based Screening of Target Genes Regulated by Heat Shock Factor AtHsfA1a in Arabidopsis thaliana

[ Objective] This study aimed to screen target genes regulated by heat shock factor AtHsfAla in Arabidopsis thaliana. [ Method] Using AtHsfAla-in- serted mutant athsfala （SALK-068042） and wild-type A. thaliana seedlings as experimental materials, target genes regulated by heat shock factor AtHsfAla were screened by microarray assay. Differentially expressed genes were screened by multiple method. Specific functions of differentially expressed genes were analyzed by gene ontology （GO） analysis. Signal transduction pathways, in which differentia｜ly expressed genes were involved, were analyzed by pathway analysis. Gene-gene interaction network was constructed by Signal-Net. [ Result] A total of 3 672 differentially expressed genes were screened out. Up-regulated differentially expressed genes were involved in 198 functions and 7 signal transduction pathways; down-regulated differentially expressed genes were involved in 94 functions and 10 signal transduction pathways. In the signal transduction network, it was found that cwlNV4 and HXK3 had relatively high ability of mediation; AT1 G14240 and cwlNV4 ex- hibited the most interactions with other genes, which were located in key positions throughout the gene-gene interaction network. [ Conclusion] Heat shock factor AtHsfAla regulates a large number of target genes in A. thaliana.

Effects of temperature on the respiratory metabolism,feeding and expression of three heat shock protein genes in Anadara broughtonii

Anadara broughtonii is one of the main commercially important species of shellfish in northern China.A.broughtonii lives in relatively stable subtidal zone where the clam could respond to environmental changes with minimum energy.Therefore,slight fluctuations in water environment may have a great impact on physiological processes such as feeding and metabolism.Large-scale mortality often occurs during the intermediate rearing processes,and high temperatures in summer are considered the leading cause of mortality.To understand the physiological and molecular response patterns of A.broughtonii at high temperatures and to estimate the appropriate metabolism temperature for A.broughtonii,the effects of temperature on the respiratory metabolism and food intake at different growth stages were studied.The response patterns of heat shock protein genes to heat stress and post-stress recovery were also explored.Results show that the temperature and growth stage of A.broughtonii were two important factors affecting metabolism and feeding.The optimum temperature for feeding and physiological activities in each shell-length group was 24℃.The temperature at which the peak metabolic rate occurred in each shell-length group increased with the increase in shell length.With the increase in heat stress,the expression of three heat shock protein genes(Abhsp60,Abhsp70,and Abhsp90)in the tissues of two size groups of A.broughtonii increased significantly when temperature was above 24℃and reached a peak at 30℃.After heat shock at 30℃for 2 h,the expression of the three heat shock protein genes rapidly increased,peaked at 2 h after the heat shock,and then gradually decreased to the level of the control group at 48 h after the heat shock.The three heat shock protein genes respond rapidly to heat stress and can be used as indicators to the cellular stress response in A.broughtonii under an environmental stress.

The polymorphism of heat shock protein 70 genes in Chinese Han population in Fujian province

To understand the polymorphism of the heat shock protein 70 （HSPTO） genes in Chinese Han population and to explore the co-relations between HSP70 polymorphism and disease, three polymorphic loci of HSP70 genes in 127 healthy Chinese Han population in Fujian province were analyzed by PCR and restriction enzyme analysis, and the genotypes and allele frequencies of HSPTO in different populations from various area were compared. It was found that the proportions of HSPTO-1 genotypes GG, GC and CC among Chinese Han population in Fujian province were 55.1%, 40.2% and 4.7% respectively, while those of HSP70-2 genotypes AA, AG and GG were 44.1%, 48.8 % and 6.9% respectively, and those of HSP70-hom genotypes TF, TC and CC were 59.8%, 37.0% and 3.2% respectively. The allele frequencies of G and C in HSP70-1 were 75.2% and 24.8% ; those of A and G in HSP70-2 were 68.5% and 31.5% and those ofT and C in HSP70-hom were 78.3% and 21.7% respectively. The distribution of the HSPTO-1 polymorphisms in Chinese Han population was almost the same as those in Japanese and Mexican populations, but it was rather different from those of American and Spanish populations with a significant differences. Meanwhile, the frequency of GG homozygote in HSPTO- 1 was signifi- cantly higher than those in American and Spanish populations. No significant difference was found in the distribution of HSPTO-2 polymorphism between Chinese and Japanese populations, in which the differences among American, Mexican and Spanish populations were quite obvious. The frequency of AA homozygote in HSPTO-2 was significantly higher than those in Mexican, American and Spanish populations, while, the distribution of HSPTO-hom genotype and allele frequency in Chinese Han population was almost just the same as those in Japanese and Mexican populations. Furthermore, it was also found that the genotype distribution and allele frequencies of the HSPTO genes in Han population of Fujian province were almost the same as those in Han population in Taiwan, but they were different in certain loci from those of Han population in Wuhan area. It is evident that the distribution of HSPTO gene polymorphisms among Chinese Han population are different from other regions in the world.

Proteomics of Wheat Endosperm:a Tool to Find Genes Involved in Kernel Composition and Quality

The composition of the wheat kernel is the result of the expression of thousands of genes translated in enzymes involved in all the biochemical pathways that are needed for endosperm cell functions and also for the ac-

Expression of Foreign Gene in Mycobacterium Regulated by Human Mycobacterium Tuberculosis Heat Shock Protein 70 Promoter

The DNA fragments of 150bp length promoter 0f human Mycobacterium(M.) tuberculosis heat shock protein (hsp)7O and 650bp length foreign gene, the Schistosoma japonicum glutathione S-transferase(Sj26GST)gene,were obtained by amplification with polymerase chain reaction. And the 150p DNA sequence upstream initiation codon ATG of the human M. tuberculosis hsp7O promoter that contains the sequence TTGAG and ATCATA which consensus with E. coli promoter's -35 and-10 region respectively, as well as ribosome binding site GGAGG at position-12-8 upstream the ATG were determined by SangerDideoxyribonucleotide-mediated chain-termination method-Then, the human M. tuberculosis hsp70 promoter and Sj26GST cDNA were cloned into E. coli-mycobacteria shuttle plasmid pBCG-2000 to construct E. coli-Mycobacterium expression shuttle plasmid pBCG- Sj26 that can express Sj26GST gene.The M. smegmatis were electroporated and the positivecolonies were selected by kanamycin-The M.smegmatis containing the vector pBCG-Sj26 can be induced by heating and hydrogen peroxide (H2O2) to express GST. The molecular weight of the recombinant GST(rGST) was 26000. The rGST contents that were about 10 percent of the total bacterial protein were analyzed by density scanning after running SDS-PAGE. This study would provide scientific evidences for application of hsp70 promoter in expressing foreign gene in mycobacterium and development of mycobacterium as multiple-valent vectoral vaccine.

Regulation of gene expression in chickens by heat stress

High ambient temperatures are a critical challenge in the poultry industry which is a key producer of the animal-based food.To evaluate heat stress levels,various parameters have been used,including growth rates,blood metabolites,and hormones.The most recent advances have explored expression profiling of genes that may play vital roles under stress.A high ambient temperature adversely affects nutrient uptake and is known to modulate the expression of genes encoding for sodium-dependent glucose transporters,glucose transporters,excitatory amino acid transporters,and fatty acidbinding proteins which are responsible for the absorption of macronutrients in the intestine.Various defensive activities are stimulated to protect the cell of different tissues from the heat-generated stress,including expression of early stress response genes coding for heat shock protein(HSP),c-FOS like protein,brain-derived neurotrophic factor(BDNF),and neuronal nitric oxide synthase(nNOS);antioxidant enzyme genes such as superoxide dismutase(SOD),catalase(CAT),and nicotinamide adenine dinucleotide phosphate oxidase(NOX4);and immune-related genes such as cytokines and toll-like receptors(TLRs).The potential role of HSPs in protecting the cell from stress and their presence in several tissues make them suitable markers to be evaluated under heat stress.BDNF and c-FOS genes expressed in the hypothalamus help cells to adapt to an adverse environment.Heat causes damage to the cell by generating reactive oxygen species(ROS).The NOX4 gene is the inducer of ROS under heat stress,which is in turns controlled by antioxidant enzymes such as SOD and CAT.TLRs are responsible for protecting against pathogenic attacks arising from enhanced membrane permeability,and cytokines help in controlling the pathogen and maintaining homeostasis.Thus,the evaluation of nutrient transporters and defense mechanisms using the latest molecular biology tools has made it possible to shed light on the complex cellular mechanism of heat-stressed chickens.As the impacts of heat stress on the above-mentioned aspects are beyond the extent to which the reduced growth performance could be explained,heat stress has more specific effects on the regulation of these genes than previously thought.

Effect of the C.–1388 A>G polymorphism in chicken heat shock transcription factor 3 gene on heat tolerance

Heat stress is one of the main factors that inlfuence poultry production. Heat shock proteins (HSPs) are known to affect heat tolerance. The formation of HSPs is regulated by heat shock transcription factor 3 (HSF3) in chicken. A DNA pool was established for identifying single nucleotide polymorphisms (SNPs) of the chicken HSF3, and 13 SNPs were detected. The bioinformatic analysis showed that 8 SNPs had the capacity to alter the transcription activity of HSF3. The dual luciferase report gene assay showed that there was a signiifcant difference (P<0.01) in the Firelfy luciferase/Renil a luciferase ratio (F/R) of C.–1 703 A>G (S1) and C.–1 388 A>G (S4) sites at the 5′-untranslated region (UTR) of chicken HSF3. The elec-trophoretic mobility shift assay showed that the S4 site was a transcription binding factor. The analysis of the association of the S1 and S4 sites with heat tolerance index revealed that the S4 site was signiifcantly correlated with the CD3+T cel , corticosterone, and T3 levels in Lingshan chickens and with the heterophil/lymphocyte value in White Recessive Rock. These results showed that the S4 site at the 5′ UTR of chicken HSF3 might have an impact on heat tolerance in summer and could be used as a potential marker for the selection of chicken with heat tolerance in the future.

Establishment of a tetracycline-off and heat shock-on gene expression system in tobacco

The tetracycline （Tet）-off gene expression regulation system based on the TetR-VP16/Top10 construct has not been widely utilized in plants, for its highly expressed TetR-VP16 activator is toxic to some plants and repeatedly replenishing tetracycline to turn off the constitutively active system is a tedious process. To solve these problems, a Tet-off and heat shock （HS）-on gene expression regulation system was constructed in this study. This system is composed of a chimeric transactivator gene TetR-HSF that is derived from a Tet repressor （TetR） and a HS transcription factor （HSF） controlled by a HS promoter HSP70m, and a Tet operator containing hybrid promoter, Om35S, that drives expression of the β-glucuronidase （GUS） gene. The resultant system yields a GUS expression pattern similar to that of the HSP70m promoter under inducing temperatures and at 35 and 40℃ drives GUS expression to a similar level as the Cauliflower mosaic virus （CaMV） 35S promoter. Further examination revealed that the TetR-HSF and GUS genes were induced by HS, reaching peak expression after 1 and 6 h treatment, respectively, and the HS induction of the expression system could be inhibited by Tet. This system will provide a useful tool for transgenic studies of plants in the laboratory and in the field, including transgene function analysis, agronomic trait improvement, biopharmaceutical protein production and others.

Characterization of a TaJ Gene from Wheat

A novel J-domain protein gene was cloned from wheat （Triticum aestivum L.） using RT-PCR technology and named as TaJ. The J-domain protein is defined by the presence of a J-domain. The cDNA of T. aestivum gene, TaJ （GenBank accession number： DQ789026）, was 1263 bp and contained a complete open reading frame （ORF） encoding a J-domain protein of 420 amino acid residues. The predicted amino acid sequence of TaJ possesses three functionally essential domains： the Nterminal J-domain which includes the highly conserved HPD tripeptide, an adjacent domain that is rich in glycine and phenylalanine residues （G/F） and a Cysteine-rich zinc-finger domain with four repeats of CxxCxGxG that is important for protein interactions. The C-terminal of TaJ was -CAQQ, a farnesylation motif. The full-length deduced amino acid sequence of TaJ is highly homologous to J-domain proteins from various plant species. Southern blot analysis indicated that a single copy of TaJ existed in wheat genome. The expression pattern of TaJ performed by real-time PCR demonstrated that heat shock （HS） at 37℃ induced the expression of TaJ rapidly and strongly, but the response of the TaJ gene to cold stress was much slower than that to HS. Tissue-specific expression analysis showed that the expression level of TaJ gene was much higher in leaves than that in roots.

Bioinformatics analysis of microarray data to explore the key genes involved in HSF4 mutation-induced cataract

AIM： To reveal the mechanisms of heat-shock transcription factor 4 （HSF4） mutation-induced cataract.METHODS： GSE22362, including 3 HSF4-null lens and 3 wild-type lens, was obtained from Gene Expression Omnibus database. After data preprocessing, the differentially expressed genes （DEGs） were identified using the limma package. Based on Database for Annotation, Visualization and Integrated Discovery （DAVID） tool, functional and pathway enrichment analyses were performed for the DEGs. Followed by protein-protein interaction （PPI） network was constructed using STRING database and Cytoscape software. Furthermore, the validated microRNA （miRNA）-DEG pairs were obtained from miRWalk2.0 database, and then miRNA-DEG regulatory network was visualized by Cytoscape software. RESULTS： A total of 176 DEGs were identified in HSF4-null lens compared with wild-type lens. In the PPI network, FBJ osteosarcoma oncogene （FOS）, early growth response 1 （EGR1） and heme oxygenase （decycling） 1 （HMOX1） had higher degrees and could interact with each other. Besides, mmu-miR-15a-5p and mmu-miR-26a-5p were among the top 10 miRNAs in the miRNA-DEG regulatory network. Additionally, mmu-miR-26a-5p could target EGR1 in the regulatory network. CONCLUSION： FOS, EGR1, HMOX1, mmu-miR-26a-5p and mmu-miR-15a-5p might function in the pathogenesis of HSF4 mutation-induced cataract.

Heat Stress Related Gene Expression in Gossypium hirsutum L.

Abiotic stress is a major limiting factor to crop productivity,and heat stress is one of the important elements for reduced crop production.Plants respond to heat stress at molecular and cellular levels as well as physiological level.Heat stress alters expression patterns of numerous genes in plants.

Cloning and Expression Analysis of Heat Shock Protein Gene DpHsp70 in Dahlia

[Objectives]This study was conducted to investigate the relationship between HSPs and the response of high temperature stress.[Methods]Molecular biological techniques were applied to clone and analyze the gene sequence of DpHsp70 gene.The changes in the expression of DpHsp70 gene under high temperature stress and exogenous salicylic acid(SA)were observed and further analyzed by qRT PCR.[Results]The coding region of the Dahlia DpHsp70 gene was 705 bp,encoding 234 amino acid residues(GenBank accession number:MH102288).Aligned with Compositae plants,the Dahlia DpHsp70 gene shared more than 83%homology in gene sequence while 99%-100%homology in amino acid sequence.Under the 35℃high temperature stress,the expression of DpHsp70 gene in Dahlia petals significantly increased.Meanwhile,the expression of DpHsp70 gene further increased under SA at 35℃temperature,which was significantly higher than those of the control group and the 35℃high temperature stress treatment group.It was demonstrated that the Hsp70 gene in Compositae is with highly conservative property and its expression could be up-regulated in response to high temperature stress.It can also be concluded that applying exogenous SA can improve the high temperature resistance of Dahlia.[Conclusions]This study provides a new experimental basis for elucidating the physiological function and mechanism of Dahlia in response to high temperature stress.

THE ROLE OF INTRONIC HSE IN THE HEAT SHOCK INDUCED EXPRESSION OF HUMAN hsp 90β GENE

The first intron of human hsp90β gene is not only essential in maintaining high constitutive expression but also critical for heat shock inducibility of the hsp90β gene. Typical HSEs in the first intron play a vital role in the heat induced expression of human hsp90β gene. Slot blot analysis shows that hsp90β gene mRNA transcripts initiated from the 3’ of the first intron dominates over that of the first exon. The intronic HSEs of the hsp90β gene show much higher binding affinity toward the recombinant heat shock factor HSF1 than that of the recombinant heat shock factor HSF2.

Transcriptomic Responses of Garlic(Allium sativum L.)to Heat and Drought Stresses

Heat and drought are prominent abiotic stressors that limit crop productivity and yield,particularly concerning climate change;therefore,understanding the molecular mechanisms underlying plant stress responses is crucial for stress-tolerant crop production.This study conducted a transcriptomic analysis to elucidate how garlic(Allium sativum L.)responds to drought and heat stress conditions.Transcriptome libraries were generated to identify differentially expressed genes(DEGs)induced by drought and heat stresses.Functional classification and clustering analysis of DEGs revealed stress-specific gene expression patterns.Notably,cell wall-related genes were implicated in the drought response,whereas heat stress was associated with heat stress transcription factors and heat shock proteins.Our results provide essential information for future studies on stress tolerances in garlic.

芝麻热胁迫响应基因SiHsfB-2b的克隆及表达特性分析

为探究B类热激转录因子(HSFs)在芝麻耐热性中的分子功能,基于前期转录组分析鉴定到的编码B类HSF家族蛋白HsfB2b的基因序列,以郑太芝3号耐热白芝麻品种为材料,采用同源克隆方法克隆了芝麻SiHsfB-2b基因,对其进行生物信息学分析,同时分析其在芝麻不同器官和热胁迫下的表达特性,并对其进行亚细胞定位。结果表明:SiHsfB-2b基因CDS序列全长为936 bp,编码311个氨基酸;系统进化树分析表明SiHsfB-2b与松蒿、独脚金和锈鳞木樨榄的同源蛋白关系较近,含保守DNA结合结构域;亚细胞定位于细胞核内;SiHsfB-2b基因启动子区域含有13类顺式作用元件,其中与胁迫响应和激素响应相关的各有3类。RT-qPCR分析结果显示,热胁迫可诱导SiHsfB-2b在芝麻根、茎和叶中上调表达,叶中相对表达量最高;在持续高温胁迫下,SiHsfB-2b在芝麻茎和叶中的表达量呈上升趋势,且在胁迫后期的叶中表现出显著上调,而根中的表达量呈现出先上升后下降趋势。综上推测,SiHsfB-2b基因可能通过介导胁迫和激素信号等通路积极响应热胁迫,这可为后期深入研究芝麻耐热性的分子机制奠定理论基础。

急性冠脉综合征患者血清sST2及NLRP3水平与介入术后无复流-慢血流的相关性分析

目的探讨急性冠脉综合征(acute coronary syndrome,Acs)患者血清可溶性生长刺激表达基因蛋白2(soluble growth stimulation expression gene 2 protein,sST2),核苷酸寡聚化结构域样受体热蛋白结构域相关蛋白3(nucleotide oligomerization domain like receptor heat protein domain associated protein 3,NLRP3)水平与经皮冠状动脉介入治疗(percutaneous coronary intervention,PCI)术后无复流-慢血流的关系。方法选择2020年1月~2022年12月佳木斯市中心医院收治的97例急性冠脉综合征患者,所有患者均接受PCI治疗,根据术后无复流-慢血流发生情况分为无复流-慢血流组(n=20)和对照组(n=77)。术前检测血清sST2及NLRP3水平,分析影响急性冠脉综合征患者PCI术后无复流-慢血流的因素以及sST2,NLRP3预测急性冠脉综合征患者PCI术后无复流-慢血流的价值。结果无复流-慢血流组血清sST2(14.32±2.65 ng/ml vs 11.02±2.13 ng/ml),NLRP3(68.23±10.17 pg/ml vs 42.05±8.23 pg/ml)水平高于对照组,差异具有统计学意义(t=5.860,12.055,均P<0.05)。多因素Logistic回归分析显示高血栓负荷(OR:7.791,95%CI:2.834~21.421)、高水平sST2(OR=2.071,95%CI:1.146~3.743)、高水平NLRP3(OR=2.008,95%CI:1.228~3.284)是急性冠脉综合征患者PCI术后无复流-慢血流的危险因素(均P<0.05)。sST2,NLRP3诊断急性冠脉综合征患者PCI术后无复流-慢血流的临界值分别为12.91ng/ml,55.39 pg/ml,曲线下面积分别为0.737,0.686,联合sST2,NLRP3诊断急性冠脉综合征患者PCI术后无复流-慢血流的曲线下面积为0.907,高于单独诊断(Z=2.662,2.856,均P<0.05)。结论急性冠脉综合征患者血清sST2,NLRP3水平增高与PCI术后无复流-慢血流的发生有关,联合检测sST2和NLRP3可提高对术后无复流-慢血流的诊断效能。

热休克蛋白90对hERG-A561V通道蛋白转运及通道功能影响的研究

目的探讨热休克蛋白90(Hsp90)对人类ether-a-go-go相关基因(hERG)-A561V通道蛋白转运及通道功能影响的研究。方法构建野生型(WT)、突变型(A561V)、杂合型(WT/A561V)细胞模型,采用蛋白质印迹法检测各组细胞hERG及Hsp90蛋白表达差异。转染减表达空载(Itf NC)、Hsp90减表达(Hsp90-)、过表达空载(Ovp NC)或Hsp90过表达(Hsp90+)质粒至各细胞模型(即Itf NC组、Hsp90-组、Ovp NC组、Hsp90+组),另设不加载体的对照(Ctl)组,采用蛋白质印迹法检测各组细胞调控Hsp90前后hERG及Hsp90蛋白表达差异。采用免疫荧光法检测Hsp90调控前后杂合型细胞模型hERG及Hsp90蛋白定位表达情况。采用全细胞膜片钳技术检测调控Hsp90前后杂合型细胞模型hERG通道激活电流及尾电流密度。结果突变型、杂合型细胞模型Hsp90及hERG(135 kDa)蛋白表达水平均较野生型明显升高(均P<0.05)。野生型细胞模型Hsp90+组hERG(135 kDa)、hERG(155 kDa)蛋白表达水平较Ctl组、Ovp NC组均明显升高(均P<0.05),突变型细胞模型Hsp90+组hERG(135 kDa)蛋白表达水平较Ctl组、Ovp NC组均明显升高(均P<0.05),杂合型细胞模型Hsp90+组hERG(155 kDa)蛋白表达水平较Ctl组、Ovp NC组均明显升高(均P<0.05)。融合Hsp90和hERG蛋白后,Ctl组hERG表达于细胞膜、细胞质,Hsp90-组细胞膜上hERG蛋白表达减少,Hsp90+组细胞膜上hERG蛋白表达增多。与Ctl组比较,Hsp90+组Ikr曲线左移14.709,斜率因子k值未见明显变化。杂合型细胞模型Ctl组、Hsp90-组、Hsp90+组激活电流及尾电流密度均在50 mV处达到最高;杂合型细胞模型Hsp90+组在20、30 mV处的激活电流密度均明显高于Ctl组、Hsp90-组(均P<0.05),在30、40 mV处的尾电流密度均明显高于Ctl组、Hsp90-组(均P<0.05)。结论Hsp90在hERG-A561V通道蛋白折叠转运中起作用,而上调Hsp90可促进WT/A561V hERG的折叠转运,进而影响通道功能。