钙结合蛋白S100A8和S100A9的异源二聚体体外构建

目的体外构建S100A8和S100A9的异源二聚体（S100A8/S100A9）,为其分子生物学功能研究提供依据。方法原核表达载体pET28 a-S100A8和pET28 a-S100A9分别转化感受态细胞BL21,并用异丙基硫化-β-D半-乳糖苷（IPTG）诱导表达蛋白,N i-NTA H is柱纯化蛋白,将等量纯化蛋白在一定钙离子浓度下室温孵育30 m in,通过SDS-PAGE和W estern b lot方法验证异源二聚体的形成。结果质粒pET28 a-S100A8和pET28 a-S100A9在相对分子质量约11 kD和14 kD处出现新生的蛋白条带,可溶性分析发现两个蛋白均可以以可溶的形式存在,纯化后得到了目的蛋白,S100A8和S100A9室温孵育后形成了异源二聚体。结论 S100A8和S100A9蛋白可以在体外形成异源二聚体。     

冠状动脉粥样硬化患者外周血单个核细胞钙结合蛋白S100A8和S100A9基因表达的变化

目的:探讨外周血单个核细胞钙结合蛋白S100A8和S100A9基因表达水平与冠状动脉粥样硬化的关系。方法:稳定性心绞痛(CAD)组61例:经冠状动脉造影(CAG)发现左主干有≥50%狭窄,或在左前降支、左回旋支和右冠状动脉主支发现至少有一处病变狭窄≥75%;对照组53例:有胸痛症状或有异常心电图、异常超声心动图表现,但冠状动脉造影表现为冠状动脉管壁光滑、未见明显粥样硬化斑块的患者。CAG前获得空腹静脉血进行血常规和高敏C反应蛋白(hs-CRP)检测、提取总核糖核酸并合成互补脱氧核糖核酸用于实时定量逆转录多聚酶链反应(RT-PCR)。结果:CAD组糖尿病比例(34.4%比15.1%,P=0.032)、合并闭塞性动脉硬化和(或)缺血性脑卒中病史比例(19.7%比0%,P=0.0006)以及hs-CRP水平[0.20比0.10,P=0.022]均高于对照组。CAD组淋巴细胞百分比低于对照组(27.2%比31.7%,P=0.019)。CAD组S100A8和S100A9基因表达水平均高于对照组[分别为0.92(0.83-1.03)比0.84(0.72-0.99),P=0.028;1.09(0.91-1.41)比0.92(0.65-1.25),P=0.009]。S100A8和S100A9基因表达水平与白细胞分类计数水平均不存在相关性(均P0.05)。糖尿病患者和无糖尿病患者的S100A8和S100A9基因表达水平无统计学意义(均P0.05)。S100A8和S100A9基因表达水平均与hs-CRP水平相关(分别为r=0.52,P=0.0000;r=0.62,P=0.0000)。结论:稳定性心绞痛患者的外周血单个核细胞S100A8和S100A9基因表达水平升高,提示其持续高表达与冠状动脉粥样硬化有关。     

血清S100A8、S100A9水平与小儿难治性肺炎支原体肺炎病情严重程度及预后的关系

目的 探讨血清S100钙结合蛋白A8(S100A8)、S100钙结合蛋白A9(S100A9)水平与小儿难治性肺炎支原体肺炎（RMPP）病情严重程度及预后的关系。方法 选取70例RMPP患儿为RMPP组、80例普通肺炎支原体肺炎患儿为GMPP组、50例健康体检儿童为对照组。根据病情严重程度将RMPP患儿分为轻症组（n=36）、重症组（n=34）,根据预后将RMPP患儿分为预后良好组（n=43）和预后不良组（n=27）。用酶联免疫吸附实验检测血清S100A8、S100A9并比较各组二者水平变化。分析S100A8、S100A9水平与RMPP患儿病情严重程度的关系;Logistic回归分析RMPP患儿预后不良的危险因素;绘制受试者工作特征曲线,用曲线下面积评价血清S100A8、S100A9水平对RMPP患儿预后的预测效能。结果 RMPP组血清S100A8、S100A9水平均高于GMPP组和对照组（P均<0.05）,GMPP组血清S100A8、S100A9水平均高于对照组（P均<0.05）。治疗前及治疗后7 d,重症组血清S100A8、S100A9水平均高于轻症组（P均<0.0...     

不同Hp感染的老年慢性胃炎患者S100蛋白表达及意义

目的 探讨不同幽门螺旋杆菌(Hp)感染情况的老年慢性胃炎患者S100蛋白表达特点及意义。方法 选取老年慢性胃炎患者92例，根据病理检查结果分为慢性非萎缩性胃炎组39例，慢性萎缩性胃炎组53例。接受碳13尿酸呼气试验，并结合快速尿素酶试验结果进行综合判断，分为Hp感染阳性(阳性组)、Hp感染阴性(阴性组)。胃镜下随机采集胃窦部黏膜组织标本，免疫组化法检测S100A8、S100A9阳性表达情况；RT-PCR检测S100A8、S100A9 mRNA表达水平；Western印迹检测S100A8、S100A9蛋白表达水平。结果 慢性萎缩性胃炎组S100A8、S100A9表达阳性率明显高于慢性非萎缩性胃炎组(P<0.01)。慢性萎缩性胃炎组S100A8、S100A9 mRNA和蛋白表达水平明显高于慢性非萎缩性胃炎组(P<0.01)。Hp感染阳性的老年慢性胃炎患者S100A8、S100A9表达阳性率明显高于Hp感染阴性的患者(P<0.01)。Hp感染阳性的老年慢性胃炎患者S100A8、S100A9 mRNA和蛋白表达水平明显高于Hp感染阴性的患者(P<0.01)。慢性萎缩性胃...     

血清S100A9、S100A2在NSCLC患者中的变化及预后的意义

目的分析血清S100钙结合蛋白A9(S100A9)、S100钙结合蛋白A2(S100A2)在非小细胞肺癌(NSCLC)患者中的变化及预后的意义。 方法选取2019年1月至2020年10月我院收治的46例NSCLC患者为对象，选择同期在我院进行健康体检的32例为对照组。对比两组血清S100A9、S100A2水平。统计NSCLC患者预后，依据预后情况分为生存者和死亡者。Logistic多因素回归分析分析影响NSCLC患者预后的因素。绘制工作特征曲线(ROC)，以曲线下面积(AUC)分析血清S100A9、S100A2联合检测对NSCLC患者预后。 结果观察组血清S100A9、S100A2水平均高于对照组(P<0.05)。截止随访结束，病死率为26.08%。Logistic回归分析结果，临床分期为Ⅲ期、分化程度为低分化、血清S100A9及S100A2水平均为影响NSCLC患者预后的危险因素(OR＝2.824、2.790、3.401、3.102，P<0.05)。ROC结果显示，血清S100A9、S100A2预测NSCLC患者预后的最佳点分别为302.11 ng/L、7.59 μg/L，两者联合的特异度为97.92%，高于血清S100A9、S100A2, S100A9、S100A2两者联合预测NSCLC患者预后的AUC为0.899，高于S100A9、S100A2单独预测的AUC(P<0.05)。 结论血清S100A9、S100A2两者联合检测对NSCLC患者预后有意义。     

S100A9蛋白的生物学特性与临床意义

S100A9蛋白属钙结合蛋白S100蛋白家族成员,通常与S100A8通过化学结合形成异二聚体,其主要功能是抑制酪蛋白激酶Ⅰ和Ⅱ的活性。近年来研究发现在多种疾病状态,包括肿瘤组织中有S100A9高表达,这提示S100A9有可能作为临床诊断的新指标。     

S100A8/A9在心脏骤停心肺复苏后脑损伤中的意义分析

目的探讨分析S100A8/A9在心脏骤停心肺复苏后脑损伤中的意义。方法选取2017年7月-2018年7月我院急诊科收治的心脏骤停心肺复苏术成功后仍需进行脑复苏患者50例,作为研究组;根据CPC评分标准,将患者分为预后差组(CPC 3-5级)和预后良好组(CPC 1-2级);同时留取同期50例来我院正常健康体检人群的血清作对照组。检测各组的血清S100A8/A9水平,并进行分析、对比。结果研究组血清S100A8/A9水平显著高于对照组(P<0.05)。预后良好组血清S100A8/A9水平显著高于预后差组及对照组(P<0.05)。结论给予心脏骤停心肺复苏脑损伤患者监测血清S100A8/A9水平,能够帮助临床医生更好地评估患者的病情转归及发展情况,为临床治疗提供更多参考依据。     

结核分枝杆菌通过诱导宿主产生S100A8/A9促进T细胞死亡的机制研究北大核心CSCD

目的探讨结核分枝杆菌(Mtb)诱导宿主产生的S100钙结合蛋白A8/A9复合物(S100A8/A9)促进T细胞死亡的作用及机制。方法灭活毒性Mtb H37Rv(iMtb H37Rv)与野生型(WT)小鼠全脾细胞体外共同培养不同时间,通过流式细胞术得到S100A8与S100A9的表达差异以及产生S100A8/A9的细胞群。Mtb H37Rv体外诱导WT和S100a9^(-/-)小鼠全脾细胞,通过流式细胞术检测S100A8/A9对CD4^(+)T细胞死亡的影响。采用不同浓度S100A8/A9与人T淋巴细胞系Jurkat细胞共同培养、S100A8/A9与细胞共培养不同时间或是S100A8、S100A9单体以及S100A8/A9复合物与细胞共同培养,通过流式细胞术得到S100A8/A9对Jurkat细胞死亡的影响。通过Western blot检测不同细胞表面TLR4的表达差异。封闭Jurkat细胞表面TLR4受体后与S100A8/A9共同培养,通过流式细胞术得到Jurkat细胞死亡的变化。结果与对照组相比,iMtb H37Rv能诱导脾细胞产生更多的S100A8和S100A9蛋白。S100A8/A9能促进小鼠CD4^(+)T细胞死亡。低剂量的S100A8/A9复合物能诱导Jurkat细胞死亡,且S100A8和S100A A9单体无以上作用。S100A8/A9复合物诱导T细胞死亡为TLR4非依赖通路。结论Mtb通过诱导宿主产生S100A8/A9并促进T细胞死亡。     

血清S100B、S100A6、S100P水平升高与急性冠脉综合征的相关性研究

目的:探讨血清终末糖基化产物受体(RAGE)的配基S100B、S100A6、S100P水平与急性冠脉综合征(ACS)相关性。方法:连续收集882例冠状动脉造影患者,检测血清S100B、S100A6、S100P、游离RAGE(sRAGE)、肿瘤坏死因子(TNF)-α水平;根据临床表现及实验室资料,将患者分为对照组(n=251)、稳定型心绞痛(SA)组(n=211)及ACS组(n=420)。结果:ACS组血清S100B、S100A6、S100P和TNF-α水平[(103.73±56.90)ng/L、(5.28±4.15)μg/L、(8.73±7.96)μg/L、(87.82±39.30)ng/L]均显著高于SA组[(81.93±27.65)ng/L、(4.36±2.45)μg/L、(3.41±3.08)μg/L、(71.88±30.70)ng/L]和对照组[(78.00±22.71)ng/L、(3.97±2.57)μg/L、(3.38±2.74)μg/L、(57.07±27.23)ng/L,P0.01];ACS组血清sRAGE水平高于对照组[(724.01±320.37)ng/L对(652.55±351.24)ng/L,P0.01]。进一步将ACS组分为ST段抬高型心肌梗死(STEMI)和不稳定型心绞痛/非ST段抬高型心肌梗死(UA/NSTEMI)2个亚组进行分析,STEMI亚组的S100B、S100A6、S100P水平高于UA/NSTEMI亚组;ACS组血清S100B水平与肌钙蛋白I(cTnI)峰值水平相关(P0.05),血清S100P水平与肌酸激酶同工酶(CK-MB)及cTnI峰值水平均有相关性(P0.01)。多元回归分析发现,S100B、S100A6、S100P和传统危险因素均与ACS发病相关。结论:血清S100B、S100A6、S100P水平与ACS相关,与心肌缺血的损伤程度相关,它们在ACS的病理生理过程中发挥重要作用。     

S100A4基因表达与胃癌生物学行为的关系

目的 探讨胃癌组织中S100A4基因表达与胃癌生物学行为的关系.方法 收集45例进展期胃癌手术切除标本,以RT-PCR法检测病灶中S100A4 mRNA的表达.按不同临床病理因素和TNM分期进行分组,比较不同组间S100A4 mRNA的表达水平.结果 S100A4 mRNA表达水平与胃癌的大体类型、浸润深度(P＜0.01)及淋巴结转移程度(P＜0.05)密切相关.TNM Ⅲ/Ⅳ期病例的S100A4表达水平明显高于Ⅰ/Ⅱ期病例(P＜0.01).结论 S100A4表达与胃癌生物学行为密切相关,其高表达是胃癌高侵袭能力和预后不良的标志之一.     

The value of mRNA expression of S100A8 and S100A9 as blood-based biomarkers of inflammatory bowel disease

Background and study aimsNon-invasive biomarkers of inflammatory bowel diseases (IBD) are of critical importance. Here, we evaluated the S100A8 and S100A9 mRNA expression, as the heterodimers of calprotectin, in the blood leucocytes of IBD patients to find how their expression associates with the disease characteristics.Patients and methodsIn this cross-sectional study, 59 IBD patients and 30 healthy subjects were included. The flare and remission phases of disease were identified in 46 and 13 patients, respectively. Blood leucocytes were isolated, and the S100A8 and S100A9 mRNA expression were evaluated in the isolated leucocytes using relative quantification real-time PCR.ResultsThe mean S100A8 and S100A9 mRNA expression were significantly higher in IBD patients than in the controls (p = 0.03 and p = 0.02, respectively). The mean S100A8 and S100A9 mRNA expression were significantly higher in the flare phase of the disease compared with the remission phase (p = 0.01 and p = 0.007, respectively). S100A8 distinguished IBD patients from controls with the sensitivity and specificity of 73% and 64%, and flare phase of disease from remission with the sensitivity and specificity of 67% and 62%. On the other hand, S100A9 distinguished IBD patients from controls with the sensitivity and specificity of 81% and 70%, and flare phase of disease from remission with the sensitivity and specificity of 68% and 64%.ConclusionThe S100A8 and S100A9 mRNA are differentially expressed in blood leucocytes of IBD patients compared to healthy controls as well as active versus quiescent disease. Thus, they can be potentially used as a blood-based biomarker in the monitoring of IBD.     

外周血钙卫蛋白、环氧化酶-2水平变化与动脉血栓性疾病的关系

目的 探讨外周血钙卫蛋白（S100A8/A9）、环氧化酶-2（COX-2）水平与动脉血栓性疾病的关系及其临床意义.方法 选取动脉血栓性疾病患者101例,其中急性心肌梗死患者50例,冠心病患者51例.25例健康体检者为对照组.采用酶联免疫吸附法（ELISA）检测各组血清S100A8/A9、COX-2水平,并分析S100A8/A9、COX-2水平间的相关性.结果 （1）动脉血栓性疾病患者外周血S100A8/A9、COX-2水平明显高于对照组;（2）急性心肌梗死组患者S100A8/A9、COX-2水平明显高于冠心病组;（3）急性心肌梗死组和冠心病组S100A8/A9水平与COX-2水平均无明显相关性.结论 心血管病患者外周血S100A8/A9、COX-2水平明显升高可能与疾病的发生、发展相关;S100A8/A9、COX-2可作为判断动脉血栓性疾病严重程度的参考指标之一.     

S100A1 in cardiovascular health and disease: Closing the gap between basic science and clinical therapy

Calcium (Ca²⁺) signaling plays a major role in a wide range of physiological functions including control and regulation of cardiac and skeletal muscle performance and vascular tone [1] and [2]. As all Ca²⁺ signals require proteins to relay intracellular Ca²⁺ oscillations downstream to different signaling networks, a specific toolkit of Ca²⁺-sensor proteins involving members of the EF-hand S100 Ca²⁺ binding protein superfamily maintains the integrity of the Ca²⁺ signaling in a variety of cardiac and vascular cells, transmitting the message with great precision and in a temporally and spatially coordinated manner [3], [4], [5] and [6]. Indeed, the possibility that S100 proteins might contribute to heart and vascular diseases was first suggested by the discovery of distinctive patterns of S100 expression in healthy and diseased hearts and vasculature from humans and animal heart failure (HF) models [7], [8], [9], [10], [11], [12], [13], [14], [15], [16], [17] and [18]. Based on more elaborate genetic studies in mice and strategies to manipulate S100 protein expression in human cardiac, skeletal muscle and vascular cells, it is now apparent that the integrity of distinct S100 protein isoforms in striated muscle and vascular cells such as S100A1, S100A4, S100A6, S100A8/A9 or S100B is a basic requirement for normal cardiovascular and muscular development and function; loss of integrity would naturally lead to profound deregulation of the implicated Ca²⁺ signaling systems with detrimental consequences to cardiac, skeletal muscle, and vascular function [7], [8], [9], [10], [11], [12], [13], [14], [15], [16], [17], [18], [19] and [20]. The brief debate and discussion here are confined by design to the biological actions and pathophysiological relevance of the EF-hand Ca²⁺-sensor protein S100A1 in the heart, vasculature and skeletal muscle with a particular focus on current translational therapeutic strategies [4], [21] and [23]. By virtue of its ability to modulate the activity of numerous key effector proteins that are essentially involved in the control of Ca²⁺ and NO homeostasis in cardiac, skeletal muscle and vascular cells, S100A1 has been proven to play a critical role both in cardiac performance, blood pressure regulation and skeletal muscle function [4,21,23]. Given that deregulated S100A1 expression in cardiomyocytes and endothelial cells has recently been linked to heart failure and hypertension [4,21,23], it is arguably a molecular target of considerable clinical interest as S100A1 targeted therapies have already been successfully investigated in preclinical translational studies.     

Alcohol intoxication inhibits pulmonary S100A8 and S100A9 expression in rats challenged with intratracheal lipopolysaccharide

BACKGROUND: Alcohol is known to inhibit the recruitment of polymorphonuclear leukocytes (PMNs) into tissue sites including the lung. During infection and inflammation, recruited neutrophils (PMNs) release S100 proteins that function to promote the recruitment of additional phagocytes. METHODS AND RESULTS: This study investigated the effects of alcohol intoxication on S100 protein production in the lung in response to lipopolysaccharide (LPS). Animals were administered alcohol (5.5 g/kg) or saline 30 minutes before intratracheal challenge with LPS (100 microg/rat). Alcohol suppressed PMN recruitment into the lung following intratracheal LPS, which was associated with an inhibition of increase in S100A8 levels in both the bronchoalveolar lavage (BAL) fluid and lysates of cells recovered by BAL at 90 minutes and 4 hours post-LPS challenge. S100A8 and S100A9 mRNA expression in cells recovered by BAL was significantly up-regulated at both 90 minutes and 4 hours after the LPS challenge, and alcohol also suppressed this response. In addition, intratracheal LPS caused a transient increase in S100A8 mRNA expression in circulating leukocytes at 90 minutes after the challenge. Similarly, this LPS-induced up-regulation of S100A8 mRNA expression was inhibited in rats intoxicated with alcohol. CONCLUSION: These data show that alcohol inhibits the S100 protein response in the lung, which may serve as a mechanism underlying alcohol-induced suppression of PMN recruitment into the terminal airways during pulmonary infection.     

S100A8 enhances IL-1β production from nasal epithelial cells in eosinophilic chronic rhinosinusitis

BackgroundChronic rhinosinusitis is classified into eosinophilic chronic rhinosinusitis (ECRS) and non-eosinophilic chronic rhinosinusitis (NECRS). ECRS is a refractory allergic disease involving a variety of immune and epithelial cells. S100A8 is a damage-associated molecular pattern that is closely related to allergic inflammation. However, the pathological implications of S100A8 in ECRS have not been clarified.MethodsWe evaluated the role of S100A8 in the pathogenesis of ECRS. Gene expression profiles of nasal polyps obtained from patients with ECRS or NECRS were evaluated using RNA sequencing.ResultsS100A8 was identified as a significantly upregulated gene in nasal polyps associated with ECRS. Immunohistochemistry consistently revealed intense S100A8 staining in nasal polyps from patients with ECRS. Human nasal epithelial cells expressed the receptor for advanced glycation end products and Toll-like receptor 4. Recombinant S100A8 protein induced interleukin-1β secretion in human nasal epithelial cells.ConclusionsOur data demonstrate that S100A8 results in production of interleukin-1β in the nasal epithelium, which may be involved in the pathogenesis of ECRS.     

Protéines S100A8, S100A9 et S100A12 : marqueurs inflammatoires ou acteurs physiopathologiques de la polyarthrite rhumatoïde

Although S100 proteins represent 40% of the neutrophil cytoplasmic proteins, their physiological and pathological functions are still unclear. S100A8, S100A9 and S100A12 protein concentrations are dramatically enhanced in synovial fluid and synovium of patients suffering from rheumatoid arthritis. Their expression seems to correlate with disease activity and joint damage. These proteins are likely involved in rheumatoid arthritis pathogenesis by enhancing extracellular matrix proteolysis, autoimmunity and inducing the pseudotumoral phenotype of the synoviocytes in rheumatoid arthritis. S100A8, S100A9 and S100A12 assessment will probably constitute a relevant tool for rheumatoid arthritis diagnosis and will improve inflammatory arthritides management.     

胰腺癌S100A4癌基因及ppENK抑癌基因的甲基化状态

目的 分析胰腺癌及胰腺癌细胞株ppENK、S100A4基因的甲基化状态.方法 收集31例胰腺癌及相应癌旁组织、5株胰腺癌细胞株及1例正常胰腺组织.采用MSP方法分析ppENK基因甲基化状态,采用COBRA方法分析S100A4基因甲基化状态,RT-PCR检测ppENK和S100A4 mRNA,免疫组化法检测S100A4蛋白表达,并与胰腺癌临床参数进行相关性分析.结果 正常胰腺组织ppENK基因无甲基化,S100A4基因高甲基化.31例胰腺癌组织中ppENK基因甲基化率为90.3%,与临床参数无相关性;S100A4基因低甲基化率为71.0%,仅与外周血CA19-9水平呈相关性(P=0.011,OR=0.05).S100A4蛋白在胰腺癌组织中表达率为87.1%,该表达与S100A4基因低甲基化相关(P=0.017),同时与肿瘤的分化程度有关,肿瘤分化程度越低,表达阳性率越高.S100A4基因低甲基化与ppENK基因甲基化相关(P=0.019).5株胰腺癌细胞株ppENK基因均甲基化,ppENK mRNA不表达;S100A4基因均低甲基化,S100A4 mRNA均高表达.结论 胰腺癌组织ppENK基因为高甲基化状态,而S100A4基因为低甲基化状态.     

EF手型钙结合蛋白S100A6及其与肿瘤关系的研究进展

S100A6蛋白是一类具有EF手型模序结构的钙结合蛋白,通过对钙离子的调节及与靶蛋白的相互作用,启动细胞信号转导,参与细胞生长、分化、细胞周期及胞外基质分泌等多种生物学过程,与肿瘤发生、发展及转移预后相关.本文就近年来S100A6钙结合蛋白的研究现状与进展进行综述.     

S100A6基因沉默对胰腺癌细胞侵袭的影响

目的 探讨S100A6基因沉默对胰腺癌细胞侵袭的影响和可能机制。方法 将不同浓度(3.125、6.25、12.5 nmol/L)的靶向S100A6的小干扰RNA( S100A6-siRNA)转染人胰腺癌BxPC3细胞,分别采用荧光实时定量PCR和蛋白质印迹法检测S100A6 mRNA和蛋白的表达,采用Transwell小室检测癌细胞侵袭能力,明胶酶谱法检测基质金属蛋白酶-9(MMP-9)活性。结果 S100A6-siRNA转染组细胞的S100A6 mRNA和蛋白表达呈浓度、时间依赖性明显下调,穿膜细胞数呈浓度依赖性明显减少。12.5 nmol/L的S100A6-siRNA转染组细胞转染后48 h的S100A6 mRNA表达从对照组的(100±0.3)％降到(15.3±0.2)％(P＜0.01);S100A6蛋白的表达从(83.2±0.18)％降到(13.5±0.12)％(P＜0.01);穿膜细胞数从(44.5±2.2)个降到(7.6±1.5)个(P＜0.05)。同时,S100A6-siRNA转染组细胞的MMP-9活性明显下调。结论 抑制S100A6基因表达可抑制胰腺癌细胞侵袭转移,其机制可能与下调MMP-9活性有关。