Effects of serum containing natural cerebrolysin on glucose-regulated protein 78 and CCAAT enhancer-binding protein homologous protein expression in neuronal PC12 cells following tunicamycin-induced endoplasmic reticulum stress

BACKGROUND： Glucose-regulated protein 78 （GRP78）, a marker of endoplasmic reticulum stress, can prolong cell survival. Alternatively, CCAAT enhancer-binding protein homologous protein （CHOP）, a transcription factor specific for endoplasmic reticulum stress, can cause cell cycle arrest and cell apoptosis. OBJECTIVE： To study the protective effects of serum containing natural cerebrolysin on endoplasmic reticulum stress in tunicamycin-induced neuronal PC12 cells, and analyze the influence on GRP78 and CHOP expressions. DESIGN, TIME AND SETTING： A parallel controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine, Shenzhen Hospital, Southern Medical University, between March 2006 and August 2008. MATERIALS： Adult Sprague-Dawley rats were perfused with natural Cerebrolysin aqueous extract （0.185 g/kg/d） to produce serum containing natural Cerebrolysin. Physiological saline was used to produce blank serum. PC12 cell line was provided by Shanghai Institute of Cell Biology, Chinese Academy of Science. Tunicamycin was provided by Sigma （St. Louis, USA）, and natural Cerebrolysin, containing ginseng, rhizoma gastrodiae, and gingko leaf （1：2：2）, by Shengzhen Institute of Integrated Western and Traditional Chinese Medicine. METHODS： PC12 cells were treated with DMEM culture media containing 10% blank serum （normal control group）, tunicamycin （1 μg/mL; model group）, and 5%, 10%, and 15% serum containing natural cerebrolysin and tunicamycin （1 μ g/mL; low-, moderate-, and high-dose serum containing natural cerebrotysin groups）, for 2 hours. MAIN OUTCOME MEASURES： PC12 cells were treated with tunicamycin for 48 hours after which apoptosis was measured using the TUNEL method to calculate apoptotic index. GRP78 expression was detected using immunocytochemistry. After 24 hours of treatment with tunicamycin, GRP78 and CHOP mRNA expressions were measured using RT-PCR. RESULTS： The apoptotic index and CHOP mRNA expression were in the model group and three cerebrolysin groups were significantly increased when compared to the normal control group （P 〈 0.05）. In contrast, GRP78 mRNA and protein expressions were significantly decreased （P 〈 0.05）. CONCLUSION： Serum containing natural cerebrolysin significantly reduced apoptosis in neuronal PC12 cells following tunicamycin-induced endoplasmic reticulum stress. These results may be related to an up-regulation of GRP78 expression and down-regulation of CHOP expression, both of which displayed dose-dependent effects.

Prostaglandin E1 protects hepatocytes against endoplasmic reticulum stress-induced apoptosis via protein kinase A-dependent induction of glucose-regulated protein 78 expression

AIM To investigate the protective effect of prostaglandin E1(PGE1) against endoplasmic reticulum(ER) stressinduced hepatocyte apoptosis, and to explore its underlying mechanisms.METHODS Thapsigargin(TG) was used to induce ER stress in the human hepatic cell line L02 and hepatocarcinomaderived cell line Hep G2. To evaluate the effects of PGE1 on TG-induced apoptosis, PGE1 was used an hour prior to TG treatment. Activation of unfolded protein response signaling pathways were detected by western blotting and quantitative real-time RTPCR. Apoptotic index and cell viability of L02 cells and Hep G2 cells were determined with flow cytometry and MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium] assay. RESULTS Pretreatment with 1 μmol/L PGE1 protected against TG-induced apoptosis in both L02 cells and Hep G2 cells. PGE1 enhanced the TG-induced expression of C/EBP homologous protein(CHOP), glucose-regulated protein(GRP) 78 and spliced X box-binding protein 1 at 6 h. However, it attenuated their expressions after 24 h. PGE1 alone induced protein and m RNA expressions of GRP78; PGE1 also induced protein expression of DNA damage-inducible gene 34 and inhibited the expressions of phospho-PKR-like ER kinase, phosphoeukaryotic initiation factor 2α and CHOP. Treatment with protein kinase A(PKA)-inhibitor H89 or KT5720 blocked PGE1-induced up-regulation of GRP78. Further, the cytoprotective effect of PGE1 on hepatocytes was not observed after blockade of GRP78 expression by H89 or small interfering RNA specifically targeted against human GRP78.CONCLUSION Our study demonstrates that PGE1 protects against ER stress-induced hepatocyte apoptosis via PKA pathwaydependent induction of GRP78 expression.

Role of Glucose-regulated Protein 78 in Early Brain Injury after Experimental Subarachnoid Hemorrhage in Rats

Early brain injury（EBI） plays a key role in the pathogenesis of subarachnoid hemorrhage（SAH）. This study investigated the role of glucose-regulated protein 78（GRP78） in EBI after SAH. Male Sprague-Dawley rats（n=108） weighing 260±40 g were divided into control, sham-operated, and operated groups. Blood was injected into the prechiasmatic cistern of rats in the operated group. Neurological scores, ultrastructures of neurons, apoptosis, and GRP78 expression in the hippocampus were examined using Garcia scoring system, transmission electron microscopy, terminal deoxynucleotidyl transferase-mediated d UTP nick-end labelling, and Western blotting at 1, 6, 12, 24, 48, and 72 h after SAH, respectively. The results showed that neurological scores were significantly decreased in the operated group as compared with those in control and sham-operated groups at 12, 24, 48, and 72 h. Metachromatin, chromatin pyknosis at the edge, endoplasmic reticulum swelling, and invagination of nuclear membrane were observed at 24 h in the operated group, indicating the early morphological changes of apoptosis. The number of apoptotic cells was significantly increased in the operated group as compared with that in control and sham-operated groups at 6, 12, 24, 48, and 72 h. The GRP78 protein expression levels in the operated group were significantly elevated at all time points and reached the peak at 12 h. GRP78 expression was positively associated with apoptosis cells and negatively with neurological scores. In conclusion, EBI was demonstrated to occur after SAH and GRP78 was involved in the development of EBI after SAH.

Glucose-regulated protein 78 inhibits scavenger receptor A-mediated internalization of acetylated low density lipoprotein

Class A scavenger receptor(SR-A) plays an important role in foam cell formation.However, the mechanism underlying the internalization of the receptor-ligand complexes remains unclear.The aim of the present study was to investigate the molecular mechanism to regulate SR-A-mediated intracellular lipid accumulation in macrophages A pull-clown assay was performed and glucoseregulated protein 78(GRP78) was identified to bind with the cytoplasmic domain of SR-A(CSR-A).Immunoprecipitation and artificially expressed protein binding assay demonstrated the direct specific binding of GRP78 with SR-A in cells.Indirect immunofluorescence assay and western blot analysis showed their co-localization in membrane and cytoplasm.Over-expression of GRP78 specifically inhibited SR-A-mediated uptake of fluorescent acetylated low-density lipoprotein, a specific ligand for SR-A, without altering cellular SR-A expression and binding ability, and significantly inhibited cholesterol ester accumulation in cells, which can be partly attributed to the suppression of c-Jun-NH2-terminal kinase signaling pathway.These results suggest that GRP78 may act as an inhibitor of SR-A-mediated internalization of modified low-density lipoprotein into macrophages(C) 2009 Elsevier Inc.All rights reserved.

Expression and significance of heat shock protein 70 and glucose-regulated protein 94 in human esophageal carcinoma

AIM: To investigate the expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human esophageal carcinoma and adjacent normal tissues.METHODS: The expression of HSP70 and grp94 in 78human esophageal cancer and adjacent normal tissues was studied by immunohistochemistry and pathology photograph analysis.RESULTS: Both esophageal cancer and adjacent normal tissues could express HSP70 and grp94. Of the 78 cases of esophageal carcinoma, 95.0%(72/78) showed positive HSP70, mainly stained in nuclei, while grp94 was mainly stained in cell plasma, and the positive rate was 71.8%(56/78).There was a significant difference in the expression of HSP70 and grp94 between esophageal cancer and adjacent normal tissues (P＜0.01). Compared with adjacent normal tissues, there was a significant difference between differential types and HSP70 expression (P＜0.01).CONCLUSION: HSP70 and grp94 express differently in cell plasma and nuclei. The expression intensity of HSP70is related to the differentiation of esophageal carcinoma.

Correlation between clinicopathology and expression of heat shock protein 70 and glucose-regulated protein 94 in human colonic adenocarcinoma

AIM: To investigate the correlation between dinicopathology and expression of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human colonic carcinoma.METHODS: The expression of HSP70 and grp94 was studied in 80 human colonic cancers with or without metastasis as well as in their adjacent mucous membrane by way of immunohistochemistry and pathology photograph analysis.RESULTS: The expression of HSP70 and grp94 was significantly higher in cancer than that in adjacent mucous membrane (92.5%, 85.0% vs 56.3%, 42.5%, P＜0.01).HSP70 and grp94 expressed higher in moderately- and poorly-differentiated colonic cancers than that in their adjacent tissues (93.7%, 87.5%; 100%, 90% vs 56.3%,42.5%; P＜0.01). Dukes C and D stages of colonic cancers showed higher positive rates than Dukes A and B stage groups (97.1%, 91.2%; 100%, 90.9%; vs 80%, 70%;78.6%, 71.4%; P＜0.05). There were definite differences in HSP70 and grp94 expression between metastasis groups and non-metastasis groups (100% vs75%, 100%vs 50%, P＜0.05).CONCLUSION: The HSP70 and grp94 expression rates in colonic cancer groups are significantly higher than that in their adjacent mucous membrane. The HSP70 and grp94expression in poorly-differentiated colonic cancers with metastasis is significantly higher than well-differentiated cancers without metastasis. The overexpression of HSP70and grp94 can be used as diagnostic or prognostic markers for colonic cancer.

Co-expression of heat shock protein 70 and glucose-regulated protein 94 in human gastric carcinoma cell line BGC-823

AIM: To investigate the co-expression and significance of heat shock protein 70 (HSP70) and glucose-regulatedprotein 94 (grp94) in human gastric carcinoma cell line BGC-823.METHODS: The expression and localization of HSP70 and grp94 in human gastric carcinoma cell line BGC-823 were determined by immunocytochemistry and indirect immunofiuorescence cytochemical staining. Flow cytometry was used to analyze the correlation between expression of HSP70, grpg4 and cell cycle in BGC-823 cell line.RESULTS: Gastric cancer cell line BGC-823 expressed high level of HSP70 and grp94. The positive rate of HSP70 and grp94 was 84.9±4.94% and 79.6±5.16%, respectively. Bothof them were stained in cell plasma. There was a significant difference compared with control group (1.9±0.94%,P＜0.01). During the cell cycle, HSP70 and grp94 were continuously expressed in BGC-823.CONCLUSION: HSP70 and grp94 are highly expressed in human gastric carcinoma BGC-823 cells through the whole cell cycle. There is no relationship between expression of HSP70, grp94 and cell cycle.

Regulation of glucose and protein syntheses by Micrococcus luteus during the fermentation of a Nigerian rice, <i>Oryza sativa</i>variety “Igbimo”

Micrococcus luteus synthesises glucose and protein during the fermentation of a Nigerian rice. To regulate the formation of these substances, mutation was carried out with an alkylating agent: ethylmethyl sulphonate (EMS). Screening the mutants generated for the levels of the traits expressed, four major groups were obtained. These are poor, moderate, good and super producers of either glucose or protein. They produced the properties at 0 - 1.00, 1.01 to 1.99 (moderate) and 2.0 to 2.99 (good) and 3.0 and above (super) mg.mL–1 of each substance. The classes were made up of 37, 40, 20 and 3 mutants for glucose production and 13, 35, 40 and 12 mutants for protein synthesis. The wild strain bacterium made 0.86 mg.mL–1 glucose and 1.2 mg.mL–1 protein describing the M. luteus as poor glucose maker and moderate protein producer. It was also noticed that the mutation caused some variants (25%) to form more glucose than protein;the remaining 75% of the population are made up of two sets viz: mutants having better ability to synthesise protein at higher concentrations than glucose and those that formed about the same amounts of the substances. It thus follows that the glucose and protein productions in M. luteus are genetically based and can be regulated by genetic manipulation.

绝经后骨质疏松症患者外周血中GRP78和ATF4的表达水平及临床意义

目的探究绝经后骨质疏松症(postmenopausal osteoporosis,PMO)患者外周血中葡萄糖调节蛋白78(glucoseregulated protein 78,GRP78)和激活转录因子4(activating transcription factor 4,ATF4)的水平及临床意义。方法选取2018年12月—2021年5月石家庄市第三医院创伤骨科收治的96例PMO患者为PMO组,另选取同期96名体检健康的绝经女性为正常组。比较PM O组和正常组的一般资料;采用酶联免疫吸附法测定血清GR P78、ATF4水平;利用电化学发光免疫分析仪测定血清β-胶原特殊序列(β-Crosslaps)、Ⅰ型前胶原氨基端前肽(procollagenⅠtype N-terminal propeptide,PINP)水平;利用双能X线骨密度仪检测腰椎骨密度(bone mineral density,BMD);采用Pearson法分析PMO患者血清GRP78、ATF4水平与β-Crosslaps、PIN P、BM D的相关性及PMO患者血清GR P78表达水平与ATF4的相关性;用Logistic回归分析PMO发生的影响因素。结果PMO组患者腰臀比、BMD低于正常组(t=9.76、15.47,P均<0.001);血清GRP78、ATF4、β-Crosslaps、PINP水平高于正常组(t=18.32、16.81、8.15、14.84,P均<0.001);PMO患者血清GRP78、ATF4水平与β-Crosslaps、PINP均呈正相关(r=0.56、0.48、0.48、0.52,P<0.001、<0.001、<0.001、=0.006),与BMD呈负相关(r=-0.54、-0.44,P均<0.001);PMO患者血清GRP78水平与ATF4呈正相关(r=0.595,P<0.001);PINP、GRP78、β-Crosslaps、ATF4是影响PMO发生的危险因素(OR=2.518、2.672、2.271、2.336,P均<0.001),BMD是影响PMO发生的保护因素(OR=0.583,P<0.001)。结论PMO患者血清GRP78、ATF4水平较高,均与β-Crosslaps、PINP、BMD相关,GRP78、ATF4可能是诊治PMO的潜在靶标,测定血清GRP78、ATF4水平有助于临床防治PMO。

天香丹通过GRP78/NLRP3信号通路调控内质网应激抗动脉粥样硬化的作用机制研究

目的:基于葡萄糖调节蛋白78(GRP78)/NOD样受体蛋白3(NLRP3)信号通路探讨天香丹通过调控内质网应激抗动脉粥样硬化的作用机制。方法:将雄性载脂蛋白E基因敲除(ApoE^(-/-))小鼠随机分为模型组、阿托伐他汀组、天香丹组,每组11只,给予高脂饲料诱导动脉粥样硬化模型;另取10只雄性C57BL/6J小鼠作为正常组。于造模10周后给予相应药物治疗,连续干预12周。采用苏木精-伊红(HE)染色观法察胸主动脉病理特点;酶联免疫吸附试验(ELISA)检测血清白细胞介素1β(IL-1β)、白细胞介素18(IL-18)、活性氧(ROS)含量;免疫组织化学法(IHC-P)检测胸主动脉中GRP78和NLRP3蛋白阳性表达水平。结果:各实验组存活6只。与模型组比较,阿托伐他汀组和天香丹组IL-1β、IL-18、ROS表达含量均降低(P<0.05或P<0.01);GRP78、NLRP3蛋白阳性表达均明显减少(P<0.05或P<0.01)。结论:天香丹能有效改善动脉粥样硬化性心血管疾病小鼠炎症反应,其作用机制可能与调控内质网应激GRP78/NLRP3信号通路抑制炎症小体活化、减轻动脉粥样硬化炎症损伤有关。

过表达葡萄糖调节蛋白78人乳腺癌细胞系HS578T、MDA-MB-231的构建

目的构建过表达内质网应激标志蛋白葡萄糖调节蛋白78(Glucose-regulated protein 78,GRP78)的人乳腺癌细胞系HS578T、MDA-MB-231,为探讨GRP78在乳腺癌发生发展中的作用机制提供基础。方法采用分子克隆技术构建表达GRP78的慢病毒载体pCDH-CMV-GRP78-HA-EF1-Puro质粒,采用PCR法扩增质粒,后使用EcoRⅠ、NotⅠ限制性内切酶切割质粒,对酶切产物进行测序鉴定,成功构建pCDH-CMV-GRP78-HA-EF1-Puro质粒。取对数生长期293FT细胞系(克隆分离株),在培养皿中加入pCDH-CMV-GRP78-HA-EF1-Puro混合液,培养48 h收集含GRP78慢病毒颗粒的上清液,保存备用。另取对数生长期人乳腺癌细胞系HS578T及MDA-MB-231,置于含GRP78慢病毒颗粒上清液的培养基中培养,培养48 h时在培养基中加入1∶500稀释嘌呤霉素,筛选GRP78过表达的HS578T、MDA-MB-231细胞系。加入嘌呤霉素培养48 h采用Western Blotting法检测HS578T、MDA-MB-231细胞GRP78。采用激光共聚焦显微镜对HS578T、MDA-MB-231细胞中GRP78蛋白进行定位。结果构建表达GRP78的慢病毒载体pCDH-CMV-GRP78-HA-EF1-Puro质粒。GRP78过表达的HS578T、MDA-MB-231细胞系在78 kD左右可见明显GPR78蛋白电泳条带。HS578T细胞系中存在GRP78蛋白表达,主要分布在细胞质中。结论成功构建GRP78过表达的MDA-MB-231、HS578T细胞系,GRP78主要表达于细胞质中。

溃疡性结肠炎患儿血清sB7-H3、GRP78水平及与病情、预后的关系

目的探究溃疡性结肠炎(UC)患儿血清可溶性B7同源体3(sB7-H3)、葡萄糖调节蛋白78(GRP78)水平及与病情、预后的关系。方法选取2019年8月至2020年8月在本院诊治的UC患儿78例作为研究组,选取同时期在本院进行健康体检的儿童75例作为对照组。根据研究组病情严重情况分为轻度组29例,中度组32例,重度组17例。对研究组进行3年随访,根据预后情况分为预后不良组22例,预后良好组56例。采用酶联免疫吸附法测定血清sB7-H3、GRP78及炎症因子干扰素-r(IFN-r)、白细胞介素-1β(IL-1β)、白细胞介素-33(IL-33)的水平;根据血压测量仪检测舒张压和收缩压水平。Pearson法分析血清sB7-H3和GRP78分别与炎症因子的相关性;Logistic回归分析影响UC患儿预后发生不良的因素;受试者工作特征(ROC)曲线分析血清sB7-H3和GRP78水平预测UC患儿预后发生不良的临床价值。结果研究组血清sB7-H3、GRP78、IFN-r、IL-1β、IL-33水平显著高于对照组(P<0.05)。随着UC患儿病情越严重,血清sB7-H3、GRP78、IFN-r、IL-1β、IL-33水平越高(P<0.05)。血清sB7-H3和GRP78分别与IFN-r、IL-1β、IL-33呈正相关(P<0.05)。不良组血清sB7-H3、GRP78、IFN-r、IL-1β、IL-33水平显著高于良好组(P<0.05)。血清sB7-H3、GRP78、IL-33是影响UC患儿预后发生不良的危险因素(P<0.05)。血清sB7-H3、GRP78及二者联合检测UC患儿预后发生不良的曲线下面积(AUC)为0.874、0.884、0.969,二者联合检测明显高于单独检测(Z二者联合-sB7-H3=1.878,Z二者联合-GRP78=1.931,P<0.05)。结论UC患儿血清sB7-H3、GRP78水平升高,其随病情越严重水平越高,二者联合可较好预测UC患儿预后发生不良的情况。

胸部CT影像特征联合血清GRP78和CST1对良恶性肺结节的诊断价值

目的 探讨葡萄糖调节蛋白78(GRP78)、半胱氨酸蛋白酶抑制剂1(CST1)在良恶性肺结节患者血清中的表达,以及联合胸部CT影像特征在诊断良恶性肺结节中的价值。方法 收集98例2020年1月至2021年12月在本院治疗的肺结节患者作为研究对象,根据病理学结果,将患者分为良性组(n=45)和恶性组(n=53)。比较两组血清GRP78、CST1表达水平和胸部CT影像特征;ROC曲线分析血清GRP78、CST1对恶性肺结节的诊断效能;四表格法分析胸部CT影像特征联合血清GRP78、CST1对恶性肺结节的诊断效能。结果 恶性组血清GRP78、CST1的表达水平高于良性组(P<0.05)。良性组和恶性组在分叶征、毛刺征、血管集束征、胸膜凹陷征之间存在显著差异(P<0.05)。胸部CT影像特征联合血清GRP78和CST1诊断恶性肺结节的准确率为92.86%,敏感度为96.23%,特异度为88.89%。其中三项联合的准确率高于胸部CT影像、血清CST1单独诊断(P<0.05),敏感度高于血清GRP78和CST1单独诊断(P<0.05)。结论 恶性肺结节患者血清GRP78、CST1表达水平显著高于良性患者;胸部CT影像特征联合血清GRP78和CST1在良恶性肺结节的诊断中具有较高的应用价值。

急性心肌梗死患者血清GRP78,HSP47,miR-126表达与心功能的相关性

目的 探讨急性心肌梗死患者血清葡萄糖调节蛋白78(GRP78)、热休克蛋白47(HSP47)、微小RNA-126(miR-126)表达与心功能的相关性。方法 选取医院2021年6月至2022年6月收治的急性心肌梗死患者80例为观察组,另选取同期健康体检者50例为对照组。采用酶联免疫吸附试验检测血清GRP78和HSP47水平及心肌肌钙蛋白I(cTnI)、肌酸激酶MB型同工酶(CK-MB)水平,采用实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-126表达水平,采用超声心动图测定左心室射血分数(LVEF)。采用Pearson相关系数法进行相关性分析,根据全球急性冠状动脉事件注册研究(GRACE)评分将观察组患者分为高危组(12例)、中危组(31例)、低危组(37例),建立受试者操作特征(ROC)曲线,评价GRP78,HSP47,miR-126鉴别高危险分层的诊断价值。结果 观察组患者血清cTnI和CK-MB水平均显著高于对照组(P <0.05),LVEF显著低于对照组(P <0.05)。观察组患者血清GRP78和HSP47水平均显著高于对照组(P <0.05),且低危组、中危组、高危组呈持续上升趋势(P <0.05);miR-126相对表达显著低于对照组(P <0.05),且低危组、中危组、高危组呈持续下降趋势(P <0.05)。GRP78,HSP47与cTnI,CK-MB均呈正相关(P <0.05),与LVEF均呈负相关(P <0.05);miR-126与cTnI,CK-MB均呈负相关(P <0.05),与LVEF呈正相关(P <0.05)。GRP78诊断急性心肌梗死灵敏度为50.00%,特异度为98.50%;HSP47灵敏度为91.70%,特异度为64.70%;miR-126灵敏度为75.00%,特异度为89.70%;GRP78,HSP47,miR-126 3项联合预测的曲线下面积为0.928(P <0.05)。结论 急性心肌梗死患者血清GRP78和HSP47存在异常高表达,miR-126存在异常低表达,且3项指标与心功能有一定相关性,单独或联合分析时对临床鉴别患者的风险层级均有参考价值。

CTA检查联合血清GRP78检测对心肌缺血性冠心病的诊断价值

目的探讨CT血管造影(CTA)联合葡萄糖调节蛋白78(GRP78)对心肌缺血性冠心病的诊断价值。方法选取2020年1月至2021年12月来本院进行治疗的115例心肌缺血性冠心病患者为观察组,同时选取来本院进行健康体检的志愿者100例为对照组。采用酶联免疫吸附(ELISA)法检测血清中GRP78表达水平。以冠状动脉造影作为“金标准”,分析对比CTA扫描与GRP78单一检测和联合检测对心肌缺血性冠心病的诊断效能;采用受试者工作特征(ROC)曲线评价血清GRP78对心肌缺血性冠心病发生的诊断价值。结果心肌缺血性冠心病患者血清中GRP78表达水平高于对照组(P<0.05);115例患者采用CTA扫描检查诊断为心肌缺血性冠心病患者共计95例,根据血清中GRP78水平诊断心肌缺血性冠心病患者共91例,二者联合诊断心肌缺血性冠心病患者共109例;CTA联合GRP78检测诊断心肌缺血性冠心病的准确率为89.30%,敏感度为94.78%,特异性为83.00%,高于CTA单独检测或GRP78单独检测(75.81%/81.86%、82.61%/79.13%、68.00%/85.00%),Kappa值=0.783,一致性较好。结论血清GRP78水平联合CTA扫描检测可提高心肌缺血性冠心病的诊断准确率、敏感度、特异度,具有较高的诊断价值。

血清GRP78、PHB1在急性ST段抬高型心肌梗死患者病情评估、预后预测方面的临床价值研究

目的:探究血清葡萄糖调节蛋白78(GRP78)和抗增殖蛋白1(PHB1)评估急性ST段抬高型心肌梗死患者病情及预后预测的临床价值。方法:选取2015年9月至2019年9月在安阳市人民医院接受治疗的188例急性ST段抬高型心肌梗死患者作为研究组,160例稳定型心绞痛患者作为对照组。使用全球急性冠状动脉事件注册(GRACE)风险评分将研究组患者分为低危组(79例)、中危组(72例)、高危组(37例)。采用酶联免疫吸附试验检测血清GRP78、PHB1的表达水平;Logistic回归分析急性ST段抬高型心肌梗死患者预后不良的影响因素;受试者工作特征(ROC)曲线分析血清GRP78、PHB1及二者联合对高危患者的评估效能及对患者预后不良的预测效能。结果:与对照组相比,研究组血清GRP78水平升高,且随患者病情危重程度加重而升高(P<0.05);血清PHB1水平降低,且随患者病情危重程度加重而降低(P<0.05)。血清GRP78、PHB1水平及二者联合评估高危患者的曲线下面积(AUC)分别为0.712、0.698、0.796,联合评估效能较优(P<0.05);与预后良好组比较,预后不良组患者血清GRP78水平升高(P<0.05),PHB1水平降低(P<0.05)。Logistic回归分析显示,吸烟史、左室射血分数(LVEF)、脑利钠肽(BNP)、低密度脂蛋白胆固醇(LDL-C)、肾小球滤过率估计值(eGFR)、GRP78、PHB1是急性ST段抬高型心肌梗死患者预后不良的影响因素(P<0.05)。ROC曲线分析显示,GRP78预测患者预后不良的AUC为0.738,敏感度为70.83%,特异度为76.72%;PHB1预测的AUC为0.758,敏感度为62.50%,特异度为81.03%;二者联合的AUC为0.837,敏感度为80.56%,特异度为74.14%。结论:血清GRP78、PHB1在急性ST段抬高型心肌梗死患者中异常表达,二者联合对评估急性ST段抬高型心肌梗死患者病情及预测患者不良预后具有较好效能。

血清Presepsin、GRP78水平在溃疡性结肠炎诊断、病情严重程度评估中的价值

目的观察溃疡性结肠炎(UC)患者血清可溶性白细胞分化抗原14亚型(Presepsin)、葡萄糖调节蛋白78(GRP78)水平,探讨其对诊断、病情严重程度评估的价值。方法将2019年3月至2021年3月于该院确诊为UC并住院治疗的100例患者纳入研究作为研究组。根据UC病情严重程度将研究组患者又分为轻度组(32例)、中度组(38例)和重度组(30例)。另外,选取同期于该院体检的90例表观健康者作为对照组。采用酶联免疫法测定纳入研究者血清Presepsin、GRP78水平。分析UC患者血清Presepsin、GRP78水平与患者临床资料的关系,血清Presepsin与GRP78水平间的相关性,比较不同病情严重程度UC患者的血清Presepsin、GRP78水平。采用Logistic回归分析UC发生的影响因素。采用受试者工作特征(ROC)曲线分析血清Presepsin、GRP78水平对UC患者的诊断价值。结果研究组血清Presepsin、GRP78均高于对照组(P<0.05)。Presepsin、GRP78水平与患者年龄、性别、体重指数、饮酒史无关(P>0.05),与白细胞计数(WBC)、血小板(PLT)水平有关(P<0.05)。轻度组、中度组、重度组的UC患者血清Presepsin、GRP78水平均依次升高(P<0.05)。Pearson相关分析显示,UC患者血清Presepsin与GRP78水平呈正相关(r=0.291,P<0.05)。Logistic回归分析显示,PLT、WBC、Presepsin、GRP78均是UC发生的影响独立因素(P<0.05)。ROC曲线分析结果显示,Presepsin、GRP78单独及两者联合检测诊断UC的曲线下面积(AUC)分别为0.834、0.842、0.924,两者联合诊断的效能优于血清Presepsin、GRP78单独诊断(P<0.05)。结论UC患者血清Presepsin、GRP78均呈较高水平,其水平随病情严重程度的增加而升高,两者联合检测对UC患者具有较高的诊断效能。

酚妥拉明联合重组人脑利钠肽治疗急性左心衰竭患者的疗效及对GRP78、PTX3和GDF-15的影响

目的:探讨酚妥拉明联合重组人脑利钠肽治疗急性左心衰竭患者的效果及对葡萄糖调节蛋白78(GRP78)、五聚素3(PTX3)和生长分化因子15(GDF-15)的影响。方法:采用随机数字表法将2021年石家庄市急救中心第五中心站收治的急性左心衰竭患者130例分为对照组(n=65)与研究组(n=65)。对照组患者在常规治疗基础上予以酚妥拉明治疗,研究组患者在对照组的基础上加用重组人脑利钠肽治疗。比较两组患者治疗前后的心功能指标,GRP78、PTX3和GDF-15水平;观察两组患者的临床疗效和治疗期间的不良反应。结果:研究组患者治疗总有效率为96.92%(63/65),显著高于对照组的87.69%(57/65),差异有统计学意义(P<0.05)。两组患者治疗后左心室射血分数、心脏指数和每搏输出量显著高于治疗前,且研究组患者显著高于对照组,差异均有统计学意义(P<0.05)。两组患者治疗后左心室舒张末期内径,GRP78、PTX3和GDF-15水平显著低于治疗前,且研究组患者显著低于对照组,差异均有统计学意义(P<0.05)。对照组、研究组患者的不良反应发生率比较[3.08%(2/65)vs.6.15%(4/65)],差异无统计学意义(P>0.05)。结论:酚妥拉明联合重组人脑利钠肽治疗急性左心衰竭的疗效和安全性均较高,可有效改善患者的心功能指标,降低GRP78、PTX3和GDF-15水平。

MOTS-c联合GRP78预测急性心肌梗死患者PCI术后MACE发生的价值

目的探讨MOTS-c和葡萄糖调节蛋白78(GRP78)表达水平对心肌梗死患者经皮冠状动脉介入治疗(PCI)术后主要不良心血管事件(MACE)预测价值。方法选择2019年10月至2022年10月到驻马店市中心医院进行PCI手术治疗的102例急性心肌梗死患者,所有患者均进行为期6个月随访,依据患者MACE发生情况将患者分为MACE组和非MACE组,其中MACE组患者有39例,非MACE组患者63例。应用查询病历和调查问卷等方式对两组患者的一般资料(年龄、性别、心率、体重指数、收缩压、舒张压、糖尿病史和心力衰竭病史)进行统计。应用酶联免疫吸附法对患者血清GRP78和MOTS-c水平进行检测。结果MACE组和非MACE组患者的糖尿病史、体重指数、年龄和性别差异无统计学意义(P>0.05),但是MACE组患者的收缩压和舒张压比非MACE组低(P<0.05),且MACE组患者的心力衰竭病史率和心率比非MACE组高(P<0.05)。MACE组患者的GRP78水平均比非MACE组患者高(P<0.05),而MACE组患者的MOTS-c水平低于非MACE组患者(P<0.05)。GRP78和MOTS-c预测的曲线下面积(AUC)为0.889和0.721,截断值分别为1.313、183.27μg·L^(-1),其灵敏度分别为76.48%和63.29%,特异度分别为98.21%和91.38%,两者联合诊断评估MACE的AUC为0.926,灵敏度为80.93%,特异度为97.32%。经过logistic多因素分析,GRP78>1.313μg·L^(-1)、MOTS-c≤183.27μg·L^(-1)和存在心力衰竭病史是急性心肌梗死患者PCI术后MACE发生的独立危险因素(P<0.05)。结论急性心肌梗死患者PCI术后MACE发生患者的GRP78水平升高,MOTS-c水平降低,GRP78联合MOTS-c对急性心肌梗死患者PCI术后MACE发生有预测价值。

草鱼呼肠孤病毒纤维蛋白VP56与草鱼GRP78蛋白互作诱导内质网应激

为了阐明草鱼呼肠孤病毒(GCRV)的感染机制,深入探究VP56(Ⅱ型和Ⅲ型GCRV特有的纤维蛋白)与宿主细胞蛋白的相互作用。实验通过免疫共沉淀(co-IP)发现,VP56与细胞内定位于内质网的分子伴侣蛋白葡萄糖调节蛋白78(GRP78)发生相互作用。而后,VP56与GRP78的互作通过基于远红外红色荧光蛋白mNeptune的双分子荧光互补系统(BiFC)得到验证。在VP56稳定表达的草鱼肾细胞系(CIK)中,相较于CIK细胞,内质网则形态发生巨大变化,产生肿胀、扩张、脱粒等现象,说明VP56激活内质网应激。通过对内质网应激下游转录因子的mRNA水平检测,发现VP56激活活化转录因子6(ATF6)介导的信号转导途径,激活非折叠蛋白反应。研究表明,GCRV-Ⅱ感染过程中破坏了细胞稳态、激活内质网应激。本研究为GCRV感染机制和抗GCRV病毒研究提供了新思路,揭示了一种新的病毒逃逸机制,有助于淡水养殖产业中草鱼出血病的防控。