共查询到20条相似文献，搜索用时 46 毫秒
1.
【目的】研究中药单体黄腐酚、异丹叶大黄素、去氧紫草素和毛蕊异黄酮与丁酸钠诱导猪宿主防御肽(host defense peptides,HDPs)表达的协同效果。【方法】中药单体黄腐酚、异丹叶大黄素、去氧紫草素和毛蕊异黄酮与丁酸钠单独或共同处理猪肠上皮细胞IPEC-J2和猪肺泡巨噬细胞3D4/31,通过测定HDPs的基因表达、细胞因子基因表达,以及细胞对产肠毒素性大肠杆菌(ETEC)的抑菌作用,评估中药单体与丁酸钠协同诱导HDPs表达的效果。【结果】黄腐酚与丁酸钠共同处理对HDPs的表达不表现协同作用;异丹叶大黄素与丁酸钠共同处理可协同诱导IPEC-J2细胞pEP2C和pBD3基因的表达(P＜0.01);去氧紫草素与丁酸钠共同处理可协同提高IPEC-J2细胞内pBD3和PG1-5基因的表达(P＜0.05;P＜0.01);毛蕊异黄酮与丁酸钠共同处理可协同促进IPEC-J2细胞内PG1-5、pEP2C、pBD2和pBD3基因(P＜0.05;P＜0.01)和3D4/31细胞内pBD3基因的表达(P＜0.05)。中药单体与丁酸钠共同处理对细胞因子表达无显著协作效果(P＞0.05)。除黄腐酚外,其他中药单体与丁酸钠单独或共同处理后均可提高细胞裂解产物对ETEC的抑菌能力,其中毛蕊异黄酮与丁酸钠处理对ETEC的抑制具有最佳协同效果。【结论】异丹叶大黄素、去氧紫草素和毛蕊异黄酮与丁酸钠在诱导HDPs表达、提高机体免疫和抗菌能力方面具有一定的协同作用。 相似文献
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The use of dogs as animal model for human atopic dermatitis (AD) is well known. Striking similarities in the pathogenesis of AD have been demonstrated. Similar alteration of host defense peptides (HDP) have been identified in both species. However, the ultrastructural/molecular alterations associated with HDPs secretion in AD have not been elucidated. We were able to use a multidisciplinary approach to investigate the secretion of HDP in canine skin. The contemporary use of indirect immunofluorescence, ELISA and scanning immune-electron microscopy gave fundamental insights in the pathomechanism of HDP alteration in AD. An increased intracellular expression and a reduced secretion of HDPs is present in atopic skin. An increased presence of HDPs was seen on the surface of atopic skin. These results suggested a defective secretion and an increased adhesion of HDPs to atopic corneocytes might be the reason of the reduced killing activity of HDPs in AD. 相似文献
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旨在探讨乳酸锌对猪空肠上皮细胞增殖及相关调控基因ZnT2、DMT1、IREG-1、MT1和ZIP4 mRNA表达的影响.用乳酸锌的锌浓度分别为50、100、150、200 mg·L-1的培养基培养IPEC-J2细胞,采用比色法测定分析细胞增殖变化；用实时荧光定量RT-PCR方法检测Zn T2、DMT1、IREG-1、MT1及ZIP4 mRNA表达,以TBPmRNA的表达水平作为内参对照.在细胞培养前36 h,乳酸锌对IPEC-J2细胞基本没有影响,随锌浓度递增细胞增殖幅度升高；乳酸锌处理IPEC-J2细胞后,Zn T2、DMT1、IREG-1及MT1 mRNA表达随锌浓度增高而升高,ZIP4 mRNA表达则随锌浓度增高而降低.添加乳酸锌可以促进IPEC-J2细胞增殖,上调ZnT2、DMT1、IREG-1、 MT1 mRNA表达,下调ZIP4 mRNA表达. 相似文献
4.
本研究旨在探讨姜黄素对猪轮状病毒(PRV)感染猪肠上皮细胞(IPEC-J2细胞)的抗病毒作用。以IPEC-J2细胞为试验对象,分别设置阴性对照组、感染PRV(感染复数=0.1)组和感染PRV后姜黄素(20μmol/L)处理组。在感染PRV后观察细胞病变,利用流式细胞术检测细胞内活性氧(ROS)含量,并通过实时荧光定量PCR(qRT-PCR)技术和病毒滴度测定法检测PRV在IPEC-J2细胞内的复制与增殖。结果表明：与阴性对照组相比,1)PRV感染导致IPEC-J2细胞病变,显著降低细胞活力(P<0.05),极显著上调细胞内ROS含量(P<0.01);2)姜黄素可显著抑制PRV在细胞内的复制与增殖(P<0.05);3)在PRV吸附细胞阶段添加姜黄素显著抑制了病毒的复制与增殖(P<0.05);4)姜黄素在感染前与PRV直接孵育能显著降低感染后病毒的滴度(P<0.05);5)PRV感染显著提高了细胞内黑色素瘤分化相关基因5、干扰素诱导蛋白44样蛋白抗体和干扰素β的mRNA相对表达量(P<0.05),显著降低了细胞内Toll样受体适配器分子1、线粒体抗病毒信... 相似文献
5.
Koh SY George S Brözel V Moxley R Francis D Kaushik RS 《Veterinary microbiology》2008,130(1-2):191-197
Enterotoxigenic Escherichia coli (ETEC) infections result in large economic losses in the swine industry worldwide. The organism causes diarrhea by adhering to and colonizing enterocytes in the small intestines. While much progress has been made in understanding the pathogenesis of ETEC, no homologous intestinal epithelial cultures suitable for studying porcine ETEC pathogenesis have been described prior to this report. In the current study, we investigated the adherence of various porcine ETEC strains to two porcine (IPEC-1 and IPEC-J2) and one human (INT-407) small intestinal epithelial cell lines. Each cell line was assessed for its ability to support the adherence of E. coli expressing fimbrial adhesins K88ab, K88ac, K88ad, K99, F41, 987P, and F18. Wild-type ETEC expressing K88ab, K88ac, and K88ad efficiently bound to both IPEC-1 and IPEC-J2 cells. An ETEC strain expressing both K99 and F41 bound heavily to both porcine cell lines but an E. coli strain expressing only K99 bound very poorly to these cells. E. coli expressing F18 adhesin strongly bound to IPEC-1 cells but did not adhere to IPEC-J2 cells. The E. coli strains G58-1 and 711 which express no fimbrial adhesins and those that express 987P fimbriae failed to bind to either porcine cell line. Only strains B41 and K12:K99 bound in abundance to INT-407 cells. The binding of porcine ETEC to IPEC-J2, IPEC-1 and INT-407 with varying affinities, together with lack of binding of 987P ETEC and non-fimbriated E. coli strains, suggests strain-specific E. coli binding to these cell lines. These findings suggest the potential usefulness of porcine intestinal cell lines for studying ETEC pathogenesis. 相似文献
6.
Awad WA Aschenbach JR Zentek J 《Journal of animal physiology and animal nutrition》2012,96(4):709-716
The digestive tract is a target for the Fusarium toxin deoxynivalenol (DON), a major cereal grain contaminant of animal and public health concern. Toxic effects of DON range from diarrhoea, vomiting and gastrointestinal inflammation to necrosis of several tissues. Following ingestion of contaminated food or feed, intestinal epithelial cells are exposed to a high concentration of ingested DON, potentially affecting intestinal functions. Pigs are considered to be the species most sensitive to DON toxicity. However, only few studies directly evaluated DON effects on porcine intestinal epithelial cells. Therefore, we used the porcine intestinal cell line (IPEC-J2) to assess short-term effects of DON on functional characteristics of the intestinal epithelial cells. The cytotoxic effect of DON on IPEC-J2 cells was evaluated by measuring the count of living cells and the activity of lactate dehydrogenase (LDH) released in the culture media at a DON concentration range from 0, 0.5, 2.5 and 10 μm. We demonstrated that DON at concentrations of 2.5 and 10 μm decreased significantly (p < 0.001) the cell count in a dose-dependent manner. At a concentration of 10 μm, DON caused cell damage, including rounding of cells, autolysis and cell loss from the monolayer. The mycotoxin, DON, increased LDH release into the culture medium compared with the control value. The alterations of LDH showed a good agreement with the decrease in cell count. Deoxynivalenol decreased the l-lactate concentration in the fluid supernatant of IPEC-J2 cells at 2.5 μm (p < 0.05) with a maximal effect at 10 μm of DON. To determine whether the altered lactate production may be linked to alterations of energy balance, we measured cellular ATP levels in IPEC-J2 cells. A significant decrease in ATP levels was seen at 48 h in a dose-dependent manner. It could be demonstrated that DON has a distinct cytotoxic effect on IPEC-J2 cells. 相似文献
7.
Innate immunity and host defense peptides in veterinary medicine 总被引：1，自引：0，他引：1
Linde A Ross CR Davis EG Dib L Blecha F Melgarejo T 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2008,22(2):247-265
Recent years have witnessed a surge in interest directed at innate immune mechanisms. Proper conceptualization of the key elements of innate immunity, however, is still a work in progress, because most research in immunology traditionally has been focused on components of the acquired immune response. The question of why an animal stays healthy in a world filled with many dangers is perhaps as interesting as why it sometimes surrenders to disease. Consequently, studies with an increased focus on inborn mechanisms of animal host defense may help further the development of appropriate preventative and therapeutic measures in veterinary medicine. Host defense peptides (HDPs) are central effector molecules of innate immunity, and are produced by virtually all living species throughout the plant and animal kingdoms. These gene-encoded peptides play a central role in multiple, clinically relevant disease processes. Imbalances in the expression of HDPs can lead to overt pathology in different organ systems and cell types in all species studied. In addition, HDPs are an ancient group of innate chemical protectors, which are now evaluated as model molecules for the development of novel natural antibiotics and immunoregulatory compounds. This review provides an overview of HDPs and is aimed at veterinary practitioners as well as basic researchers with an interest in comparative immunology involving small and large animal species. 相似文献
8.
IPEC-J2 cells are porcine intestinal columnar epithelial cells that were isolated from neonatal piglet mid-jejunum. This cell line forms polarized monolayers with high transepithelial electrical resistance when cultured on 0.4 μm pore-size filters. The cell line is unique in that it is derived from small intestinal tissue (compared to the common human colon-derived lines HT-29, T84, and Caco-2) and is not transformed (compared to the porcine small intestinal line, IPI-2I). Porcine intestinal epithelial cells more closely mimic human physiology than analogous rodent cell lines (e.g. IEC-6 or IEC-18), which is important in studies of zoonotic infections; in addition, they provide specificity to study porcine-derived infections. IPEC-J2 cells are increasingly being used in microbiological studies to examine the interactions of various animal and human pathogens, including Salmonella enterica and pathogenic Escherichia coli, with intestinal epithelial cells. The IPEC-J2 cell line has also been employed in some probiotic studies, in which the cells have been used as an initial screening tool for adhesiveness and anti-inflammatory properties of the potential probiotic microorganisms. The validity of these studies is not clear as follow-up studies to assess the efficacy of the probiotics in vivo have not been published to date. The aims of this review are to provide a comprehensive overview of the microbiological studies that have been conducted with IPEC-J2 cells and a reference guide of key cellular and immune markers that have been identified in this cell line that may prove to be useful in future studies. 相似文献
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Arce C Ramírez-Boo M Lucena C Garrido JJ 《Comparative immunology, microbiology and infectious diseases》2010,33(2):161-174
The innate immune system has the basic function of identifying and eradicating microbial invaders and alerting the adaptative immune system to their presence. In this study, the porcine intestinal innate immune response was evaluated by analysing the expression of TLRs, cytokines and chemokines in two porcine epithelial cell lines from different regions: IPEC-J2 (jejunum) and IPI-2I (ileum). Both cells lines were stimulated with 1microg of LPS from Salmonella typhimurium. RNA was collected at 30min, 1, 2, 3 and 4h after treatment. Expression of TLR-1, -2, -3, -4, -6, -8, -9, -10, TNF-alpha, IL-1beta, -8 and MCP-1 was quantified relative to the quantity of Cyclophilin-A mRNA using real-time quantitative PCR (RTQ-PCR). The results obtained show up differences in the gene expression between both cell lines IPEC-J2 and IPI-2I as response to LPS from S. typhimurium during the activation time, which may suggest an in vivo variability in the innate immune response against pathogens in different regions of the host's gut. 相似文献
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Volf J Boyen F Faldyna M Pavlova B Navratilova J Rychlik I 《Zoonoses and public health》2007,54(8):286-293
Salmonella enterica serovar Typhimurium (S. Typhimurium) is a facultative intracellular bacterium which can infect and colonize pigs. After contact with enterocytes and macrophages, S. Typhimurium induces production of cytokines thus triggering the innate immune response. In this study we evaluated the cytokine response of two porcine cell lines, IPI-2I and 3D4/31, of epithelial or macrophage origins, respectively, to the wild-type S. Typhimurium and its hilA and ssrA mutants. We observed that the 3D4/31 cell line essentially did not respond to S. Typhimurium infection when a medium with foetal calf serum was used. However when the 3D4 cell line was incubated overnight in the presence of porcine serum, it efficiently responded to the wild-type strain and the ssrA mutant but not to the noninvasive hilA mutant as measured by mRNA quantification of TNF-alpha, IL-8 and GM-CSF by the real-time RT-PCR. In IPI-2I, all the cytokines were also induced by the wild-type S. Typhimurium and the ssrA mutant although the induction of TNF-alpha was lower than that induced by the wild-type strain. The hilA mutant was unable to induce any of the cytokines tested. The ssrA mutant can therefore be considered as more suitable for further vaccine development as the stimulation of innate immune response is important for animal protection against a challenge with virulent strains. 相似文献
13.
Skjolaas KA Burkey TE Dritz SS Minton JE 《Veterinary immunology and immunopathology》2007,115(3-4):299-308
Direct-fed microbials, including Lactobacillus and Bacillus spp., are potential replacements for low dose in-feed antibiotics for swine and other livestock. To understand the function of these microbes in the gut, the current study used pig jejunal epithelial cells (IPEC-J2) to evaluate how Lactobacillus reuteri (LR) and Bacillus licheniformis (BL) differed from Salmonella enterica serovars Typhimurium (ST) or Choleraesuis (SC) in their ability to regulate, stimulate, or modify the proinflammatory mediators, interleukin 8 (IL8), CC chemokine 20 (CCL20), and tumor necrosis factor-alpha (TNFalpha). To optimize the positive control to drive IL8 secretion by IPEC-J2 cells, cells were treated apically with various concentrations of ST (versus control (CTL)) for 1h, followed by a wash. Media containing gentamicin was added and collected at 6h post-treatment. Compared to CTL, 10(8) ST produced maximal IL8 secretion in both the apical and basolateral directions, with significant basolateral polarization (P<0.0001). We next evaluated the time course of IL8 secretion, and IL8, CCL20, and TNFalpha mRNA expression by IPEC-J2 cells treated apically with 10(8) ST, SC, LR, and BL versus CTL. Media and RNA were collected at 1.5, 3.0, and 6.0 h post treatment. Only ST stimulated an increase in IL8 secretion at any time point, with increases in IL8 mRNA at both 3 and 6h (P<0.05). However, BL increased IL8 mRNA at 1.5h (P<0.0001). Neither LR nor SC affected IL8 mRNA expression. CCL20 mRNA was strongly upregulated by ST (P<0.05) and BL (1.5 and 3.0 h; P<0.05), but not LR or SC. Only ST increased TNFalpha mRNA relative to CTL (P<0.05). Two experiments were conducted to determine if pre-exposure of IPEC-J2 cells to LR or BL modified ST induced IL8 secretion. Confluent cells were treated apically overnight with various levels of LR or BL (in separate experiments) followed by ST challenge. Media were collected at 4 (LR experiment) or 5h (BL experiment) post ST. In the LR study, IL8 secretion was increased by ST as compared to CTL (P<0.0001), reduced by LR (P<0.05), and LR+ST co-treatments failed to alter ST stimulated secretion. In the BL experiment, secretion of IL8 was increased by ST (P<0.0001), but blunted basolaterally in BL+ST co-treated wells. The data demonstrate that IPEC-J2 cells increase IL8 secretion in response to ST, and IL8 mRNA in response to ST and BL, but not LR. Furthermore, ST stimulated secretion of IL8 is inhibited basolaterally in the presence of BL. 相似文献
14.
本研究旨在探讨表皮生长因子(EGF)调控猪小肠上皮细胞IPEC-J2中钠依赖Ⅱb型磷转运蛋白(NaPi-Ⅱb)表达的分子机制。试验分别用EGF受体酪氨酸激酶抑制剂(tyrphostin AG1478)、蛋白激酶A(PKA)抑制剂(H89)、蛋白激酶C(PKC)抑制剂(k4393)、p38抑制剂(SB203580)、细胞外信号调节激酶(ERK)抑制剂(PD98059)、c-Jun氨基末端激酶(JNK)抑制剂(anisomycin)与EGF共同处理IPEC-J2细胞,利用Western blot检测相关通路蛋白及目的蛋白(NaPi-Ⅱb)的表达水平。结果显示:相较于对照组,EGF处理后NaPi-Ⅱb表达水平显著降低(P0.05);相较于无抑制剂组,EGF受体、PKA、PKC、丝裂原活化蛋白激酶(MAPK)/p38、MAPK/ERK1/2、MAPK/JNK的特异性抑制剂处理IPEC-J2后,NaPi-Ⅱb表达水平显著提高(P0.05),其中添加MAPK/ERK1/2特异性抑制剂显著降低了MAPK/ERK1/2在Tyr204位点的磷酸化水平(P0.05),添加MAPK/JNK的特异性抑制剂显著降低了MAPK/JNK1/2/3在Thr183和Tyr185位点的磷酸化水平(P0.05),说明该2组抑制剂对该通路的抑制作用是通过降低上述位点的磷酸化水平实现的。本研究结果表明EGF受体、PKA、PKC、p38、ERK和JNK均介导了EGF调控IPEC-J2细胞中NaPi-Ⅱb的表达。 相似文献
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从河北省保定某两个猪场疑似高致病性猪繁殖与呼吸综合征病料中分离出能引起Marc-145细胞病变的病毒,经RT-PCR鉴定证明,该病毒为美洲型PRRSV,分别命名为BD1和BD2株。并将其克隆、测序,与国内外PRRSV分离株的Nsp2和E基因序列进行了比较。结果表明,与PRRSV高致病性毒株JXA1、GD、WUH1、SY0608、HUN0701的核苷酸序列同源性为95.3%~96.5%,BD1和BD2株ORF5基因存在部分突变,没有缺失,与国内高致病性PRRSV分离株JXA1、HUB1、Jiangxi-3、Henan-1、WUH1的同源性高达97.2%~99.3%。 相似文献
16.
Pavlova B Volf J Alexa P Rychlik I Matiasovic J Faldyna M 《Veterinary microbiology》2008,132(1-2):105-110
Enterotoxigenic Escherichia coli (ETEC) is an extracellular bacterium that causes post-weaning diarrhoea (PWD) in piglets with different severity of clinical signs. The pathogenesis of ETEC is ascribed to the effect of enterotoxins. ETEC colonizes ileum and probably can penetrate the epithelium and stimulate macrophages. The aim of study was to examine whether there is any difference in cytokine response in vitro produced by two porcine cell lines, intestinal epithelial cell line (IPI-2I) and macrophage cell line (3D4/31) after stimulation with different serotypes of ETEC associated with different clinical course of PWD in piglets. Three serotypes, O149:K88 (F4), O147:F18 and O8:K88, were used. We observed that all the used serotypes were unable to induce IL-8 and TNF-alpha mRNA expression in IPI-2I cell line as measured by the real-time RT-PCR. In 3D4/31 cell line, we detected differences in cytokine response among the used serotypes. The highest IL-8 and TNF-alpha mRNA expression in 3D4/31 was detected after stimulation with serotype O149:K88 frequently associated with hemorrhagic gastroenteritis. 相似文献
17.
酶制剂和微生态制剂对断奶仔猪养分利用率和生长性能影响的研究 总被引：1，自引：1，他引：0
试验分为两个部分,旨在研究添加抗生素、酶制剂和微生态制剂对仔猪养分利用率和仔猪生长性能的影响。饲养实验:选用体重基本一致的28日龄商品断奶仔猪320头,试验分为4个处理,每个处理4个重复,处理1为对照组(无抗生素、酶制剂、微生态制),处理23、、4分别在处理1的基础上添加抗生素(喹乙醇)100 mg/kg、酶制剂350 mg/kg、微生态制剂1 000 mg/kg。试验期35 d。消化试验:选用55日龄仔猪16头,随机分为4个处理,每个处理4个重复,试验采用全收粪法,日粮和设计同饲养试验。试验结果表明,酶制剂和微生态制剂与对照组相比显著地提高了仔猪蛋白表观消化率(P<0.05),与抗生素组相比差异不显著(P>0.05)。酶制剂试验组与其他各组相比,显著提高了仔猪中性洗涤纤维的表观消化率(P<0.05)。酶制剂和微生态制剂对仔猪总能和干物质的表观消化率没有显著影响(P>0.05)。酶制剂试验组和抗生素组与微生态制剂组和对照组相比显著提高了仔猪平均日增重(P<0.05),而微生态制剂组与对照组相比差异不显著(P>0.05)。微生态和酶制剂组与抗生素和对照组相比显著降低了料重比(P<0.05)。酶制剂、微生态制剂和抗生素试验组对仔猪的平均日采食量没有产生显著的作用(P>0.05),但显著降低了仔猪腹泻率(P<0.05)。酶制剂和微生态制剂可以提高仔猪养分表观消化率和生产性能。从对仔猪生产性能和养分表观消化率影响上看,酶制剂和微生态制剂可以代替抗生素应用在仔猪生产中。 相似文献
18.
本研究探索了丁酸梭菌分离株的益生特性，旨在为其在仔猪日粮中的应用提供理论依据。利用常规方法分离丁酸梭菌并进行纯培养，经生化检测和16S rRNA基因测序，对分离菌株进行鉴定；采用活菌计数法和牛津杯法研究分离株培养上清对3种致病菌的抑制作用、分离株黏附猪小肠上皮细胞（IPEC-J2）的特性及其对致病菌黏附细胞的抑制作用；采用细胞计数法研究分离株对IPEC-J2生长的影响；采用ELISA检测分离株处理细胞后培养上清液中细胞因子水平。结果表明，本研究成功分离到一株丁酸梭菌。与对照组相比，丁酸梭菌培养上清对3种致病菌生长抑制效果均显著（P<0.05）；丁酸梭菌可黏附于IPEC-J2，并且黏附效果最佳的感染复数（MOI）为50，最适处理时长为3 h；丁酸梭菌对3种致病菌黏附IPEC-J2均具有显著抑制作用（P<0.05）；丁酸梭菌MOI为1、10和50时细胞生长正常、形态完好，MOI为100时IPEC-J2生长受到显著抑制，同时出现部分细胞死亡；MOI为1时细胞因子水平无差异，MOI为10、50和100时细胞因子水平均显著增高（P<0.05）。综上所述，本研究分离的一株丁酸梭菌具有较好的益生特性，为其在生产中的应用提供了理论依据。 相似文献
19.
旨在从体内与体外探究过量赖氨酸对断奶仔猪的影响。本试验选用144头杜×长×大三元杂交断奶仔猪，随机分为4组，每组6个重复，每个重复6头仔猪（3头阉公猪和3头母猪），各组在日粮中分别添加1.3%、2.6%、3.9%和5.2%的回肠标准可消化赖氨酸（standardized ileal digestibility lysine，SID Lys），观察其对于仔猪生长性能、器官指数以及生理生化等指标的影响。以IPEC-J2为体外模型，根据培养基与营养需要中氨基酸的种类与比例，添加赖氨酸及其他氨基酸，测定IPEC-J2细胞的增殖情况，并试图通过平衡氨基酸手段降低因过量添加赖氨酸造成的不利影响。结果显示，随着日粮SID Lys添加量的增加，断奶仔猪的生长性能显著下降（P<0.05），血浆平均红细胞血红蛋白含量（mean corpuscular hemoglobin，MCH）、部分血清游离氨基酸含量和肠道形态受到影响。当在培养基中添加2.0 mmol·L-1赖氨酸时，IPEC-J2细胞活力显著下降（P<0.05），培养基中原赖氨酸浓度（0.5 mmol·L-1）是IPEC-J2生长的最适浓度。当按照培养基中氨基酸平衡比例补充其他必需氨基酸时，细胞活力有一定程度的改善；当按照猪碱性氨基酸的最适比例补充相应浓度的精氨酸与组氨酸时，细胞活力随处理时间的变化而变化。综上所述，单独添加过多的赖氨酸对断奶仔猪的生长会产生负面作用，在生产实际应用中应充分考虑氨基酸之间的平衡。 相似文献
20.
为确定猪传染性胃肠炎病毒（Transmissible gastroenteritis virus，TGEV）非结构蛋白2（nonstructural protein 2，Nsp2）与宿主细胞蛋白26S蛋白酶体非ATP酶调节亚基11（PSMD11）之间是否存在相互作用，本研究利用RT-PCR方法扩增猪源PSMD11基因，并构建其真核表达载体pCMV-Myc-PSMD11，经测序和双酶切验证正确后，转染猪小肠上皮细胞（IPEC-J2），通过Western blotting和间接免疫荧光试验（IFA）检测真核表达载体pCMV-Myc-PSMD11是否能在IPEC-J2中表达。利用免疫共沉淀（Co-IP）试验检测TGEV Nsp2和宿主细胞PSMD11蛋白之间的相互作用，并通过激光共聚焦显微镜观察TGEV Nsp2与PSMD11在宿主细胞中的共定位情况。结果显示，本研究成功扩增了猪源PSMD11基因，大小约为1 474 bp，基因序列经测序比对与标准序列完全一致。构建的真核表达载体pCMV-Myc-PSMD11能在IPEC-J2细胞中成功表达PSMD11蛋白；Co-IP结果表明，PSMD11与Nsp2之间存在相互作用；共定位试验结果显示，PSMD11与Nsp2的相互作用发生在细胞质中，且细胞中PSMD11蛋白的表达位置并未因Nsp2的表达而发生改变。本研究结果为进一步研究TGEV Nsp2在病毒感染过程中所发挥的重要作用提供新的线索。 相似文献