血管紧张素转换酶基因多态性与心肌梗死后心室重构

目的：探讨血管紧张素转换酶ACE基因I／D多态性与心肌梗死（心梗）后心功能不全和心室重构的相关性。方法：选择77名接受冠脉造影的陈旧性心梗患者，二维超声测定其左室射血分数（LVEF）和左室舒张末内径（LVDd)并采用聚合酶链反应（PCR）进行ACE I／D基因分型，分析造影和临床资料，判定心梗后心功能和心室重构与ACE I／D多态性的相关性。结果：不同基因型心梗后LVEF和LVDd均存在显著性差异（P＜0．05），其中ACE II型患者的LVEF显著性差异分别大于ID及DD型，II型患者LVDd与DD型存在显著性差异（P＝0．0505），而II型与ID型无显著性差异，结论：ACE I／D多态性可能是心梗后心功能不全及心室重构的重要先天因素。     

老年心力衰竭患者心室重构与血管紧张素转换酶基因多态性的相关性及药物干预效果的研究

目的探讨老年充血性心力衰竭(CHF)患者心室重构与血管紧张素转换酶(ACE)基因多态性的相关性,并观察美托洛尔缓释片、依那普利的干预效果。方法选择50例正常对照组和104例老年CHF患者,并将104例老年CHF患者随机分为A组(常规治疗)26例;B组(常规治疗+美托洛尔缓释片)26例;C组(常规治疗+依那普利)26例;D组(常规治疗+美托洛尔缓释片+依那普利)26例,均连续治疗3个月。所有受试着进行ACE基因分型,用放射免疫法测定AngⅡ、ALD;紫外吸收法测定ACE;PCR测定基因多态性;超声心动图测量LVEDD、LVEDV和LVEF。结果老年CHF组DD基因型频率和D等位基因频率较正常对照组明显增高,P<0.01;DD基因型患者血清ACE、AngⅡ、ALD水平明显高于ID型、II型和正常对照组,P<0.01;DD基因型较ID型、II型患者LVEDD、LVEDV明显高,而LVEF明显低,P<0.01;ACEI/D多态性与血清中ACE水平明显相关,DD型水平最高,II型最低;老年CHF伴左心室肥厚组DD基因型频率及D等位基因频率较不伴左室肥厚组明显增高,P<0.01;治疗3个月后,B组及C组中ACEDD基因型患者LVEDD、LVEDV明显低于ID型、II型及A组,而LVEF明显增高;且DD基因型患者的血清ACE、AngⅡ、ALD水平明显低于ID型、II型及A组;联合用药组DD基因型患者的LVEDD、LVEDV明显低于B组及C组,而LVEF明显增高。结论老年CHF患者血清ACE、AngⅡ、ALD水平与ACE基因多态性中DD基因型及D等位基因密切相关;老年CHF患者LVEDD、LVEDV、LVEF与ACE基因多态性中DD型及D等位基因关系密切;ACE基因多态性中DD基因型及D等位基因是老年CHF患者预后的重要预测因素,可作为判定老年CHF患者心室重构、估计病情、判断预后的重要指标之一;美托洛尔缓释片与依那普利均能改善老年CHF的心室重构及预后,但两药联合治疗效果更好,使其病死率进一步降低。     

葶苈大枣泻肺汤通过ACE2-Ang-(1-7)-Mas轴对心梗后心衰模型大鼠心室重构的影响

目的 探讨经方葶苈大枣泻肺汤对心肌梗死后心力衰竭模型大鼠心室重构的作用机制。方法 应用左冠状动脉前降支结扎术构建心肌梗死后心力衰竭大鼠模型,分8组:假手术组、模型组、A779组(1 mg/kg)、A779(1 mg/kg)+葶苈大枣泻肺汤等效剂量组(0.8 g/kg)、A779(1 mg/kg)+葶苈大枣泻肺汤高剂量组(1.6 g/kg)、葶苈大枣泻肺汤等效剂量组(0.8 g/kg)、葶苈大枣泻肺汤高剂量组(1.6 g/kg)和氯沙坦钾组(10 mg/kg),各组分别给予等体积蒸馏水或者相应的药物,灌胃4周。采用Masson染色法测定大鼠心肌组织中胶原纤维分布;采用碱水解法测定心肌组织羟脯氨酸(Hyp)含量;采用免疫组化法检测心肌组织中Ⅰ型胶原蛋白(COLⅠ)、Ⅲ型胶原蛋白(COLⅢ)的表达水平;采用酶联免疫吸附测定法检测大鼠血清中基质金属蛋白酶2(MMP-2)、MMP-9、金属蛋白酶组织抑制因子1(TIMP-1)、可溶性致瘤性抑制因子2(sST-2)等心肌纤维化相关指标;采用Western blot法检测心肌组织中血管紧张素转换酶2-血管紧张素-(1-7)-Mas[ACE2-Ang-...     

ACE2通过下调AT1R和ERK1/2的磷酸化而抑制平滑肌细胞的增殖

目的 探讨血管紧张素转换酶2(ACE2)对血管紧张素Ⅱ1型受体(AT1R)和细胞外基质激酶(ERK)1/2磷酸化水平及细胞增殖的影响.方法 荧光显微镜下观察绿色荧光蛋白(GFP)的表达;将载有ACE2基因慢病毒表达载体(Lentiviral-ACE2)以感染复数为10(MOI=10)感染平滑肌细胞不同时间(24、48、72、96 h);采用CCK-8法检测平滑肌细胞增殖;Western blot检测ACE2、AT1R及ERK1/2磷酸化水平.结果 目的 蛋白GFP表达量明显增加,慢病毒ACE2的表达量成时间依赖性增加,并且在96 h时蛋白表达量较对照组和空载体组明显升高,(1.22±0.06 vs 0.53±0.03和0.53±0.09,P<0.05,n=4);AngⅡ(10-7 mol·L-1)可以促进平滑肌细胞的增殖,ACE2能够抑制AngⅡ诱导的平滑肌细胞增殖(0.53±0.10 vs 0.68±0.03,P<0.05,n=5);AngⅡ(10-7 mol·L-1)作用平滑肌细胞12 h后AT1R明显上调,同时ACE2明显抑制AngⅡ诱导的AT1R的上调(0.34±0.01 vs 0.73±0.07,P<0.05,n=4); ACE2能够明显抑制AngⅡ诱导的ERK1/2的磷酸化水平 (0.43±0.06 vs 0.71±0.08,P<0.05,n=4).结论 ACE2可以通过下调AT1R和/及ERK1/2 的磷酸化水平而抑制平滑肌增殖,提示ACE2的过表达具有血管保护作用.     

ACE2通过下调AT1R和ERK1/2的磷酸化

目的探讨血管紧张素转换酶2(ACE2)对血管紧张素Ⅱ1型受体(AT1R)和细胞外基质激酶(ERK)1/2磷酸化水平及细胞增殖的影响。方法荧光显微镜下观察绿色荧光蛋白(GFP)的表达;将载有ACE2基因慢病毒表达载体(Lentiviral-ACE2)以感染复数为10(MOI=10)感染平滑肌细胞不同时间(24、48、72、96 h);采用CCK-8法检测平滑肌细胞增殖;Western blot检测ACE2、AT1R及ERK1/2磷酸化水平。结果目的蛋白GFP表达量明显增加,慢病毒ACE2的表达量成时间依赖性增加,并且在96 h时蛋白表达量较对照组和空载体组明显升高,(1.22±0.06 vs 0.53±0.03和0.53±0.09,P<0.05,n=4);AngⅡ(10-7 mol·L-1)可以促进平滑肌细胞的增殖,ACE2能够抑制AngⅡ诱导的平滑肌细胞增殖(0.53±0.10 vs 0.68±0.03,P<0.05,n=5);AngⅡ(10-7mol·L-1)作用平滑肌细胞12 h后AT1R明显上调,同时ACE2明显抑制AngⅡ诱导的AT1R的上调(0.34±0.01 vs 0.73±0.07,P<0.05,n=4);ACE2能够明显抑制AngⅡ诱导的ERK1/2的磷酸化水平(0.43±0.06 vs 0.71±0.08,P<0.05,n=4)。结论 ACE2可以通过下调AT1R和/及ERK1/2的磷酸化水平而抑制平滑肌增殖,提示ACE2的过表达具有血管保护作用。     

丝裂素活化蛋白激酶反义寡核苷酸对血管紧张素Ⅱ诱导的大鼠心肌成纤维细胞c-myc基因的表达及细胞增殖的抑制效应(英文)

目的:探讨丝裂素活化蛋白激酶(MAPK)反义寡核苷酸(ODN)对血管紧张素Ⅱ(Ang Ⅱ)诱导的心肌成纤维细胞c-myc基因及其细胞增殖的抑制效应。方法:MAPK反义ODN转染培养新生大鼠心肌成纤维细胞,Western Blot法结合p-81滤纸法测定MAPK活性;Northern Blot法检测c-myc mRNA的表达;[~3H]TdR掺入和[~3H]脯氨酸接入测定细胞DNA和胶原蛋白的合成。结果:MAPK反义ODN显著抑制Ang Ⅱ诱导的MAPK蛋白表达及其活性;显著抑制c-myc基因的表达以及细胞DNA和胶原蛋白的合成。结论:MAPK反义ODN特异性下调MAPK的活性,有效抑制了Ang Ⅱ诱导的c-myc基因的表达以及心肌成纤维细胞的增殖和胶原蛋白的合成。     

基因共表达网络分析及蛋白质相互作用筛选核分裂周期蛋白80与肺腺癌分期及预后的相关性

目的 通过基因共表达网络分析(weighted gene co-expression network analaysis,WGCNA)及蛋白质相互作用(protein-protein interaction,PPI)识别肺腺癌发生发展中的枢纽基因.方法 从TCGA数据库中下载497例肺腺癌组织和54例正常肺组织的RNA...     

整合生物信息学法与加权基因共表达网络分析联合分析脑胶质瘤特征基因

目的 利用脑胶质瘤（glioblastoma,GBM）芯片数据,采用整合生物信息学法和加权基因共表达网络分析（weighted gene correlation network analysis,WGCNA）法,寻找肿瘤发病特征基因、关键通路以及转录调控机制。方法 利用GEO数据库中高通量基因芯片数据,通过整合生物信息学法筛选出差异基因;利用WGCNA分析GBM关键基因hub基因;采用维恩分析法,整合这些差异基因与hub基因,筛选出GBM特征基因。采用基因本体论（Gene Ontology,GO）基因功能注释,京都基因和基因组数据库（Kyoto Encyclopedia of Genes and Genomes,KEGG）通路富集,分析GBM特征基因所富集的功能和通路。利用Kaplan-Meier分析特征基因与GBM生成率的关系。利用基因云生物信息平台（GCBI）,分析调控这些特征基因的转录因子。结果 经分析,发现273个特征基因。这些特征基因主要可影响离子通道、蛋白激酶、γ-氨基丁酸受体功能。CHD5,SYP,PHYHIP表达水平与GBM生存率显著相关;转录因子Sp1、Sp3、REST可能是调控这些特征基因的关键因子。结论 本研究从多种角度定义了GBM的特征基因及调控机制,为其精准治疗提供了依据。     

乌头碱调控miR-150-5p表达对心力衰竭模型大鼠心功能及心室重构的影响

目的 探究乌头碱对心力衰竭模型大鼠心功能及心室重构的影响及对miR-150-5p的调控作用。方法 将SPF级SD雄性大鼠随机分为假手术组、模型组、曲美他嗪组、乌头碱组、乌头碱+antagomir-NC组、乌头碱+miR-150-5p antagomir组,每组15只。除假手术组外,其余组均利用结扎左前降支冠状动脉法建立心力衰竭大鼠模型。测定各组大鼠心功能指标左室舒张末期内径(LVEDD)、左室收缩末期内径(LVESD)、左室射血分数(LVEF);ELISA检测各组大鼠心肌损伤指标心肌肌钙蛋白I(CTnI)、脑钠肽(BNP)和N末端B型脑钠肽前体(NT-pro BNP)的水平;测定各组大鼠心脏和左心室质量指数;Masson染色观察各组大鼠心肌组织形态;TUNEL染色检测各组大鼠心肌细胞凋亡率;RT-qRCR检测各组大鼠心肌组织中miR-150-5p和细胞周期蛋白D2(CCND2)的表达;双荧光素酶报告基因实验验证miR-150-5p与CCND2的靶向关系;Western blotting检测心肌细胞中CCND2蛋白的表达。结果 与模型组相比,乌头碱组大鼠心功能指标LVEDD、LVESD、...     

基于ABCB1 C3435 T基因多态性研究氯吡格雷用于急性冠状动脉综合征安全性的荟萃分析

目的:基于ABCB1 C3435T基因多态性,应用荟萃(meta)分析的方法评价急性冠状动脉综合征(acute coronary syndrome,ACS)患者使用氯吡格雷的安全性.方法:检索万方数据库、中国生物医学文献数据库、中国期刊全文数据库、中文科技期刊全文数据库(VIP)以及英文数据库Science Citat...     

Identification of immune-related lncRNAs in periodontitis reveals regulation network of gene-lncRNA-pathway-immunocyte

BackgroundLong noncoding RNAs (lncRNAs), which is essential in regulating multiple biological functions, have been found to have pivotal roles in immune regulation. Since immune reaction and immunocytes are the key part in periodontitis progression, we aim to investigate the underlying lncRNA-immunity regulatory network of periodontitis. Methods: A series of bioinformatic algorithms were used to identify immune-related lncRNAs in periodontitis. Infiltrating immunocyte were calculated by MCP-count. Pathway activity were estimated by the GSVA. The relationships between immune-related lncRNA and periodontitis features were investigated including immune gene categories, perturbated lncRNAs, immunocytes and pathways. Immune-related periodontitis subtypes were identified by ConsensusClusterPlus algorithm. Immunocytes related gene-lncRNA modules were identified by WGCNA. Results: An integrated algorithm and pipeline to identify immune-related lncRNAs was developed and 1059 immune-related lncRNAs in 14 immune categories were identified, 291 of them were perturbated in periodontitis. An independent validation set verified the robustness of immune-related lncRNAs. A higher proportion of immune-related lncRNAs are correlated with immunocyte infiltration. Pathways associated with immune-related lncRNAs were also revealed. Two distinct immune-related periodontitis subtypes were identified according to perturbated immune-related lncRNAs with different immune and clinical characteristics, in which subtype-1 has a higher infiltrated immunocytes, higher immune reaction scores and more chronic periodontitis samples. Immunocytes and clinical phenotypes matching their gene-lncRNA modules, and their functions were annotated. Conclusions: Our study systematically investigated periodontitis immune-related lncRNAs and have taken a glimpse of the underlying mechanism of periodontitis from gene-lncRNA-immunocyte networks, which can not only inspire researchers but also help in periodontitis related immune researches.     

Gene profiles of a human bronchial epithelial cell line after in vitro exposure to respiratory (non-)sensitizing chemicals: Identification of discriminating genetic markers and pathway analysis

Respiratory sensitization is a concern for occupational and environmental health in consumer product development. Despite international regulatory requirements there is no established protocol for the identification of chemical respiratory sensitizers. New tests should be based on mechanistic understanding and should be preferentially restricted to in vitro assays. The major goal of this study was to investigate the alterations in gene expression of human bronchial epithelial (BEAS-2B) cells after exposure to respiratory sensitizers and respiratory non-sensitizing chemicals, and to identify genes that are able to discriminate between both groups of chemicals. BEAS-2B cells were exposed during 6, 10, and 24 h to the respiratory sensitizers ammonium hexachloroplatinate IV, hexamethylene diisocyanate, and trimellitic anhydride, the irritants acrolein and methyl salicylate, and the skin sensitizer 1-chloro-2,4-dinitrobenzene. Overall changes in gene expression were evaluated using Agilent Whole Human Genome 4× 44K oligonucleotide arrays. Fisher Linear Discriminant Analysis was used to obtain a ranking of genes that reflects their potential to discriminate between respiratory sensitizing and respiratory non-sensitizing chemicals. The 10 most discriminative genes were BC042064, A_24_P229834, DOCK11, THC2544911, DLGAP4, NINJ1, PFKM, FLJ10986, IL28RA, and CASP9. Based on the differentially expressed genes, pathway analysis was used to identify possible underlying mechanisms of respiratory sensitization. We demonstrated that in bronchial epithelial cells the canonical PTEN signaling pathway is probably the most specific pathway in the context of respiratory sensitization. Results are indicative that the BEAS-2B cell line can be used as an alternative cell model to screen chemical compounds for their respiratory sensitizing potential.     

Identification of a novel PAK1 inhibitor to treat pancreatic cancer

Pancreatic cancer is one of the most aggressive cancers with poor prognosis and a low 5-year survival rate. The family of P21-activated kinases (PAKs) appears to modulate many signaling pathways that contribute to pancreatic carcinogenesis. In this work, we demonstrated that PAK1 is a critical regulator in pancreatic cancer cell growth. PAK1-targeted inhibition is therefore a new potential therapeutic strategy for pancreatic cancer. Our small molecule screening identified a relatively specific PAK1-targeted inhibitor, CP734. Pharmacological and biochemical studies indicated that CP734 targets residue V342 of PAK1 to inhibit its ATPase activity. Further in vitro and in vivo studies elucidated that CP734 suppresses pancreatic tumor growth through depleting PAK1 kinase activity and its downstream signaling pathways. Little toxicity of CP734 was observed in murine models. Combined with gemcitabine or 5-fluorouracil, CP734 also showed synergistic effects on the anti-proliferation of pancreatic cancer cells. All these favorable results indicated that CP734 is a new potential therapeutic candidate for pancreatic cancer.     

3D disorganization and rearrangement of genome provide insights into pathogenesis of NAFLD by integrated Hi-C,Nanopore, and RNA sequencing

The three-dimensional (3D) conformation of chromatin is integral to the precise regulation of gene expression. The 3D genome and genomic variations in non-alcoholic fatty liver disease (NAFLD) are largely unknown, despite their key roles in cellular function and physiological processes. High-throughput chromosome conformation capture (Hi-C), Nanopore sequencing, and RNA-sequencing (RNA-seq) assays were performed on the liver of normal and NAFLD mice. A high-resolution 3D chromatin interaction map was generated to examine different 3D genome hierarchies including A/B compartments, topologically associated domains (TADs), and chromatin loops by Hi-C, and whole genome sequencing identifying structural variations (SVs) and copy number variations (CNVs) by Nanopore sequencing. We identified variations in thousands of regions across the genome with respect to 3D chromatin organization and genomic rearrangements, between normal and NAFLD mice, and revealed gene dysregulation frequently accompanied by these variations. Candidate target genes were identified in NAFLD, impacted by genetic rearrangements and spatial organization disruption. Our data provide a high-resolution 3D genome interaction resource for NAFLD investigations, revealed the relationship among genetic rearrangements, spatial organization disruption, and gene regulation, and identified candidate genes associated with these variations implicated in the pathogenesis of NAFLD. The newly findings offer insights into novel mechanisms of NAFLD pathogenesis and can provide a new conceptual framework for NAFLD therapy.