血清miR-21、miR-148b、miR-200a、miR-200b在原发性肝癌患者中的表达及临床诊断意义

目的研究原发性肝癌(HCC)患者血清中微小RNA(miR)-21、miR-148b、miR-200a、miR-200b的表达及临床诊断意义。方法应用实时荧光定量逆转录-聚合酶链反应(qRT-PCR)检测西南医科大学附属医院收治的93例HCC患者(病例组)、48例肝脏良性病变(良性组)、48例慢性肝病患者(慢性肝病组)和50例健康体检者(对照组)血清中miR-21、miR-148b、miR-200a、miR-200b的表达。统计学分析各指标与临床病理特征之间的关系,受试者工作特征(ROC)曲线分析各指标的诊断价值。结果病例组血清miR-21与miR-200b表达显著高于良性组、慢性肝病组及对照组(均P<0.05),miR-200a与miR-148b表达显著低于良性组、慢性肝病组及对照组(均P<0.05)。HCC患者血清miR-21与miR-200b表达与病理分期、病理分级及肿瘤直径有关(均P<0.05),与性别、年龄、有无肝硬化及乙型肝炎病毒(HBV)抗原是否阳性无关(均P>0.05)。血清miR-200a与miR-148b表达与病理分期、病理分级有关(均P<0.05),与性别、年龄、肿瘤直径、有无肝硬化及HBV抗原是否阳性无关(均P>0.05)。miR-21、miR-148b、miR-200a、miR-200b及四项指标联合的ROC曲线下面积(AUC)分别为0.821、0.668、0.772、0.734、0.922。联合诊断的敏感性和特异性均高于任一单一指标。结论HCC患者血清miR-21与miR-200b表达上调,而miR-200a与miR-148b表达下调。并且其表达与病理分期、病理分级有关,联合检测血清miR-21、miR-148b、miR-200a、miR-200b有希望成为新的诊断HCC的肿瘤标志物。     

慢性乙型肝炎患者血清miR-21和miR-148b水平变化及其预测肝组织炎症分级和纤维化分期价值探讨*

目的 探讨应用血清miR-21和miR-148b水平预测慢性乙型肝炎(CHB)患者肝组织病理学病变的价值。方法 2016年7月～2021年6月我院诊治的CHB患者134例,行快速肝穿刺活检,评估肝组织炎症活动度分级(G)和肝纤维化分期(S)。采用实时定量逆转录PCR法检测血清miR-21和miR-148b水平。应用受试者工作特征曲线(ROC)下面积(AUC)评估血清miR-21和miR-148b水平评估CHB患者肝组织炎症分级和纤维化分期的效能。结果 经肝组织检查,在134例CHB患者中,发现肝组织炎症活动度分级包括G1级23例、G2级39例、G3级47例和G4级25例,肝纤维化分期包括S1期31例、S2期46例、S3期28例和S4期29例;G1级、G2级、G3级和G4级血清miR-21水平分别为(1.1±0.2)、(1.5±0.3)、(1.8±0.4)和(2.2±0.6),血清miR-148b水平分别为(2.4±0.5)、(2.0±0.3)、(1.6±0.5)和(1.2±0.4),组间差异有统计学意义(P＜0.05);S1期、S2期、S3期和S4期血清miR-21水平分别为(0.9±0.4)、(1.4±0.3)、(1.9±0.5)和(2.7±0.5),血清miR-148b水平分别为(2.8±0.7)、(1.9±0.4)、(1.4±0.4)和(0.9±0.2),组间差异有统计学意义(P＜0.05);分别以血清miR-21=1.5和miR-148b=2.0为截断点,其联合评估肝组织炎症分级大于或等于G2的AUC为0.875,显著高于两项指标单独评估(分别为0.769和0.781,P＜0.05),其灵敏度、特异度和准确度分别为87.0%、82.9%和83.6%;分别以血清miR-21=1.4和miR-148b=1.9为截断点,其联合评估肝纤维化分期大于或等于S2的AUC为0.898,显著高于两项指标单独评估(分别为0.782和0.770,P＜0.05),其灵敏度、特异度和准确度分别为87.1%、73.8%和76.9%。结论 利用CHB患者血清miR-21高水平和miR-148b低水平的特点可以用于评估肝组织炎症分级和肝纤维化分期,值得深入研究。     

miR-374促进急性心肌梗死诱导心肌纤维化的机制

目的研究微小RNA(miR)-374通过调控白细胞介素(IL)-10对急性心肌梗死(AMI)诱导心肌纤维化的影响。方法采用实时荧光定量-聚合酶链反应(qRT-PCR)检测AMI患者和健康对照组血清中miR-374的表达水平。构建AMI小鼠模型,随机分为假手术组(n=6)、AMI组(n=12)、AMI+miR-374组(n=12,AMI小鼠心肌注射miR-374)。采用qRT-PCR检测假手术组和AMI组心肌组织中miR-374的表达水平及各组Ⅰ型胶原(COL1)A1和Ⅲ型胶原(COL3)A1 mRNA表达水平。检测各组心脏功能,记录左室射血分数(LVEF)、左室长轴缩短分数(FS)、左室舒张末期内径(LVDd)和左室收缩末期内径(LVDs)。比色法检测各组半胱氨酸-天冬氨酸蛋白酶(Caspase)3和Caspase8活性。Massontrichrome染色检测各组心肌纤维化程度。miR靶基因数据库预测IL-10为miR-374的潜在靶基因并采用荧光素酶报告基因法验证。采用qRT-PCR检测转染miR-374 mimics对IL-10 mRNA表达的影响。采用Western印迹检测AMI组和AMI+miR-374组心肌组织中IL-10、P65和P50蛋白表达。结果AMI患者血清中miR-374的表达水平明显高于健康对照组(P<0.001)。AMI组心肌组织中miR-374表达水平明显高于假手术组(P<0.001)。相比AMI组,AMI+miR-374组LVEF和FS明显降低、LVDd和LVDs明显升高、Caspase3和Caspase8活性明显升高、COL1A1和COL3A1 mRNA表达水平明显升高、心肌纤维化程度增加(P<0.05、P<0.01、P<0.001)。miR-374可靶向调控IL-10并抑制其mRNA表达。相比AMI组,AMI+miR-374组心肌组织中IL-10蛋白表达明显降低,而P65和P50蛋白磷酸化明显增加(P<0.05)。结论miR-374可能通过抑制IL-10的表达显著促进心肌梗死诱导心肌纤维化的发展。     

miR-148 b在非小细胞肺癌患者血清中的表达水平及其预后价值分析

目的探讨miR-148b在非小细胞肺癌患者血清中的表达水平及其预后价值。方法选取本院确诊的非小细胞肺癌患者196例,同期选择门诊健康体检者48例作为对照组,采用逆转录实时荧光定量法测定血清miR-148b mRNA蛋白的表达。结果肺癌组miR-148b mRNA表达水平低于对照组,差异有统计学意义(P 0. 05);肺癌组miR-148b mRNA阳性率低于对照组,差异有统计学意义(P 0. 05); miR-148b mRNA的表达率与年龄、肿瘤最大径、肿瘤数目,相关性不明显,差异无统计学意义(P 0. 05);与病理学分级、TNM分期、淋巴血管间隙浸润、淋巴结转移、复发相关明显,且病理学分期越高、TNM分期越高、淋巴血管间隙浸润、淋巴结转移和复发患者的miR-148b mRNA阳性表达率越低,差异有统计学意义(P 0. 05); miR-148b表达阳性组3年生存率及生存期均明显高于miR-148b阴性组,差异有统计学意义(p 0. 05)。结论 miR-148b在非小细胞肺癌患者血清中的表达量低于健康人群,miR-148b在非小细胞肺癌的发生发展过程中起抑制作用; miR-148b阳性表达者可能获得较好的预后。     

miR-125b对人心肌细胞HCM的保护作用及机制

目的探讨微小RNA(miR) 125b对人心肌细胞HCM的保护作用及其可能的作用机制。方法选取急性心肌梗死(AMI)患者68例为AMI组、体检健康对照者50例作为正常对照组,采集静脉血检测血清miR-125b表达。取对数生长期人心肌HCM细胞,分别采用瞬时转染法转染miR-125b mimics(浓度分别为200、100、50 nmol/L)及NC mimics(浓度分别为200、100、50 nmol/L),培养24 h及48 h,采用CCK-8法测定细胞相对增殖率。分别转染miR-125b mimics终浓度为50、100 nmol/L,转染NC mimics终浓度为100 nmol/L及空白组,采用流式细胞术测定细胞早期凋亡百分比。采用Western blotting法检测细胞免疫、炎症相关蛋白炎性小体(NLRP3)、白细胞介素1β(IL-1β)、趋化因子2(CCL2)、趋化因子C-X3-C-配体1(CX3CL1)、补体应答基因-32(RGC32)表达。结果 AMI组血清miR-125b表达水平低于正常对照组(P <0. 01)。24、48 h时,转染各浓度miR-125b...     

miR-29b过表达对动脉硬化模型大鼠干预作用及机制

目的 探究miR-29b过表达对动脉硬化模型大鼠的干预作用及机制。方法 选取40只SD健康雄性大鼠,其中10只不进行任何处理为正常组,其余30只建立动脉硬化大鼠模型(其中10只为疾病模型组,10只进行沉默miR-29b表达干预为miR-29b沉默组,10只进行过表达miR-29b表达干预为miR-29b过表达组)。选择使用反转录(RT)-聚合酶链反应(PCR)对miR-29b表达进行检测,并以免疫组化法检测内皮生长因子(VEGF)、转化生长因子(TGF)-β1及单核细胞趋化蛋白(MCP)-1水平,酶联免疫吸附试验检测一氧化氮(NO)、内皮素(ET)-1水平,以黄嘌呤氧化酶法检测活性氧(ROS)、丙二醛(MDA)及超氧化物歧化酶(SOD)水平。结果 与正常组相比,疾病模型组、miR-29b沉默组及miR-29b过表达组miR-29b表达明显低,VEGF水平明显高(P<0.05);与疾病模型组相比,miR-29b沉默组miR-29b表达明显低,VEGF水平明显高(P<0.05),miR-29b过表达组miR-29b表达明显高,VEGF水平明显低(P<0.05)。与正常组相...     

sST2参与急性心肌梗死后心肌纤维化

目的探索可溶性生长刺激表达基因2(sST2)与急性心肌梗死(AMI)后心肌纤维化的关系。方法入选2015年1月至2016年9月入住兰州大学第一医院心脏中心的患者249例,AMI组为首次诊断AMI的166例患者,对照组为冠状动脉造影阴性的83例患者。测定患者血清sST2、Ⅲ型前胶原氨基端肽(PⅢNP)及N末端B型利钠肽原(NT-pro BNP)水平,并收集心脏超声中与左心室收缩功能相关的指标:左心室射血分数(LVEF)、左心室收缩期末容积(LVESV)、左心室舒张期末容积(LVEDV),并进行比较、分析。结果 AMI组血清sST2、PⅢNP、NTpro BNP水平及LVESV、LVEDV值均高于对照组,LVEF值明显低于对照组(P0.05或P0.01)。AMI组中,LVEF50%亚组血清sST2水平高于LVEF≥50%亚组(P=0.031)。AMI组中血清sST2与PⅢNP呈正相关(r=0.181,P=0.02),与LVEF呈负相关(r=-0.179,P=0.021)。AMI组中血清sST2、NT-pro BNP及sST2+NT-pro BNP诊断AMI后心力衰竭的ROC曲线下面积分别为0.608、0.683和0.732。结论血清sST2参与AMI后心肌纤维化过程,并与左心室收缩功能有关。血清sST2水平对AMI后心力衰竭具有诊断价值。     

miR-29 b/MMP2轴参与慢性阻塞性肺疾病的作用机制研究

目的 探究miR-29b/基质金属蛋白酶2(MMP2)轴,在慢性阻塞性肺疾病(COPD)进展中的作用.方法 采用LPS气道滴入联合烟雾法复制COPD大鼠模型,检测大鼠肺功能、血浆MMP2含量,HE观察肺组织形态学变化,qRT-PCR检测肺组织miR-29b水平;体外培养人支气管上皮样细胞16HBE,采用miRNA转染1...     

B淋巴细胞对急性心肌梗死后纤维化的作用及机制的研究进展

急性心肌梗死(acute myocardial infarction,AMI)是冠状动脉粥样斑块破裂,形成血栓,导致血管完全闭塞,造成部分心肌缺血坏死的一种严重心血管疾病[1]。虽然AMI患者经药物溶栓或急诊经皮介入治疗及时开通冠状动脉,提高AMI救治率,但仍有一部分患者发生心肌纤维化。心肌梗死后纤维化一旦进展为病理性重构,可引起严重不良心脏事件的发生。     

新活素对老年急性心肌梗死伴心力衰竭后心脏纤维化的影响及预后分析

急性心肌梗死(acute myocardial infarction,AMI)是冠状动脉粥样硬化性疾病最严重的类型,是致残致死的重要原因[1].AMI后最常见的并发症为心力衰竭(心衰),在老年人中更为常见,其也是影响老年病人预后最重要的因素[2].新活素是一种重组人脑钠肽(rhBNP),研究表明,rhBNP具有利钠、利...     

Dysregulation of microRNAs after myocardial infarction reveals a role of miR-29 in cardiac fibrosis

Acute myocardial infarction (MI) due to coronary artery occlusion is accompanied by a pathological remodeling response that includes hypertrophic cardiac growth and fibrosis, which impair cardiac contractility. Previously, we showed that cardiac hypertrophy and heart failure are accompanied by characteristic changes in the expression of a collection of specific microRNAs (miRNAs), which act as negative regulators of gene expression. Here, we show that MI in mice and humans also results in the dysregulation of specific miRNAs, which are similar to but distinct from those involved in hypertrophy and heart failure. Among the MI-regulated miRNAs are members of the miR-29 family, which are down-regulated in the region of the heart adjacent to the infarct. The miR-29 family targets a cadre of mRNAs that encode proteins involved in fibrosis, including multiple collagens, fibrillins, and elastin. Thus, down-regulation of miR-29 would be predicted to derepress the expression of these mRNAs and enhance the fibrotic response. Indeed, down-regulation of miR-29 with anti-miRs in vitro and in vivo induces the expression of collagens, whereas over-expression of miR-29 in fibroblasts reduces collagen expression. We conclude that miR-29 acts as a regulator of cardiac fibrosis and represents a potential therapeutic target for tissue fibrosis in general.     

Role of oxidative stress in cardiac remodelling after myocardial infarction

Recovery from myocardial infarction is associated with a series of alterations in heart structure and function, collectively known as cardiac remodelling, which play a major role in the subsequent development of heart failure. Early remodelling involves infarct scar formation in the ischaemic zone whereas subsequent ventricular remodelling affects mainly the viable non-infarcted myocardium with especially profound alterations in the extracellular matrix. There is growing evidence for a role of oxidative stress and redox signalling in the processes underlying cardiac remodelling. Reactive oxygen species are a group of highly reactive molecules which have the potential to modulate several biological processes as well as cause tissue damage and dysfunction. Their effects can be beneficial or deleterious, depending on the concentrations produced, the site of production, and the overall redox status of the cell. Reactive oxygen species can be generated by all cardiovascular cell types. Under pathophysiological conditions, major enzymatic sources appear to be mitochondria, xanthine oxidase and the non-phagocytic NADPH oxidases. In this review, we outline the mechanisms underlying the progression of early and late cardiac remodelling with particular focus on the role of oxidative stress and the potential sources of reactive oxygen species which may be involved.     

Molecular and cellular mechanisms involved in cardiac remodeling after acute myocardial infarction

The extent of cardiac remodeling determines survival after acute MI. However, the mechanisms driving cardiac remodeling remain unknown. We examined the effect of ischemia and reperfusion (R) on myocardial changes up to 6 days post-MI. Pigs underwent 1.5 h or 4 h mid-LAD balloon occlusion and sacrificed or 1.5 h occlusion followed by R and sacrificed at 2.5 h, 1 day, 3 days, and 6 days. Ischemic- (IM) and non-ischemic myocardium (NIM) was obtained for molecular analysis of: 1) apoptosis (P-Bcl2, Bax, P-p53, active-caspase-3); 2) the TLR-4-MyD88-dependent and independent pathways; 3) Akt/mTOR/P70^S6K axis activation; and, 4) fibrosis (TGF-β, collagen1-A1/A3). Histopathology for inflammation, collagen, and fibroblast content, TUNEL staining, and metalloproteinase activity was performed. Apoptosis is only detected upon R in IM cardiomyocytes and progresses up to 6 days post-R mainly associated with infiltrated macrophages. The Akt/mTOR/P70^s6K pathway is also activated upon R (IM) and remains elevated up to 6 days-R (P < 0.05). Ischemia activates the TLR-4-MyD88-dependent (cytokines/chemokines) and -independent (IRF-3) pathways in IM and NIM and remains high up to 6 days post-R (P < 0.05). Accordingly, leukocytes and macrophages are progressively recruited to the IM (P < 0.05). Ischemia up-regulates pro-fibrotic TGF-β that gradually rises collagen1-A1/-A3 mRNA with subsequent increase in total collagen fibrils and fibroblasts from 3 days-R onwards (P < 0.005). MMP-2 activity increases from ischemia to 3 days post-R (P < 0.05). We report that there is a timely coordinated cellular and molecular response to myocardial ischemia and R within the first 6 days after MI. In-depth understanding of the mechanisms involved in tissue repair is warranted to timely intervene and better define novel cardioprotective strategies.     

NLRP3炎性体在心肌梗死后心室重塑中的作用

NLRP3炎性体是一种细胞内多蛋白复合物,是无菌性炎症反应的关键介质,在心肌梗死（myocardial Infarction,MI）的病理生理机制中发挥重要作用。NLRP3炎性体可调节半胱氨酸蛋白酶（caspase-1）的激活,促进IL-1β等炎症因子产生,参与细胞程序性死亡。大量研究表明,抑制NLRP3炎性体可有效减轻MI后炎症反应,从而改善MI后心功能不全及心室重塑。因此,NLRP3炎性体可能是减少MI后心血管事件的发生及改善预后新的治疗靶点,本文就NLRP3炎性体在MI后心室重塑中的作用进行综述。     

Chymase inhibition prevents cardiac fibrosis and dysfunction after myocardial infarction in rats.

Human chymase activates not only angiotensin II but also transforming growth factor-beta, a major stimulator of myocardial fibrosis, while rat chymase activates transforming growth factor-beta, but not angiotensin II. To clarify the role of chymase-dependent transforming growth factor-beta activation, we evaluated whether chymase inhibition prevents cardiac fibrosis and cardiac dysfunction after myocardial infarction in rats. Myocardial infarction was induced by ligation of the left anterior descending coronary artery. One day after the ligation, rats were randomized into 2 groups: 1) a chymase-treated group that received 10 mg/kg per day of the chymase inhibitor NK3201 orally for 4 weeks; and 2) a vehicle group of non-treated rats with myocardial infarction. We also included a control group who underwent sham-operation and no treatment. Four weeks after ligation, echocardiography revealed that chymase inhibitor treatment reduced the akinetic area and increased fractional area change but did not significantly change left ventricular end-diastolic area. Chymase inhibition significantly reduced left ventricular end-diastolic pressure, increased the maximal end-systolic pressure-volume relationship and decreased the time constant of left ventricular relaxation. Chymase activity in the non-infarcted myocardium was significantly increased in the vehicle group, but it was significantly reduced by chymase inhibitor treatment. The fibrotic area in the cardiac tissues and the mRNA levels of collagen I and collagen III were also significantly lower in the chymase inhibitor-treated group than in the vehicle group. Therefore, the pathway forming chymase-dependent transforming growth factor-beta may play an important role in myocardial fibrosis and cardiac dysfunction rather than left ventricular dilatation after myocardial infarction.     

曲尼司特对心肌梗死后心肌间质纤维化的影响

目的探讨曲尼司特对兔心肌梗死后心肌间质纤维化干预作用。方法结扎左前降支制作兔心肌梗死模型,分实验组和对照组。3周后经胃管分别给予曲尼司特及安慰剂1月,心脏彩超评价心功能并检测血清转化生长因子（transform ing growth factor,TGF-β1）,I、III型胶原浓度及组织羟脯胺酸含量。结果实验组治疗前后心功能、心腔内径、室壁厚度明显改善,血清TGF-β1,I、III型胶原浓度及羟脯胺酸含量较对照组明显下降。结论曲尼司特可有效拮抗心肌梗死后心肌间质纤维化,预防心室重构。     

Mechanisms involved in cardiac enlargement and congestive heart failure development after acute myocardial infarction.

For 3 months, we followed up 40 patients with acute myocardial infarction, 20 were randomly assigned to treatment with captopril and 20 to placebo, to elucidate mechanisms inducing left ventricular volume enlargement and development of congestive heart failure. Echocardiographic follow-up could be obtained in 28 patients, 11 of whom showed more than a 10% increase in left ventricular systolic and/or diastolic volumes (captopril n = 3/15, placebo n = 8/13, p = 0.05). Volume increase was significantly associated with an impairment in exercise capacity (VO2 max in patients with vs. without volume enlargement 24.7 +/- 1.7 vs. 29.5 +/- 1.9 ml O2/kg/min; p < 0.05). Plasma renin activity, angiotensin II and catecholamines were normal in the acute and chronic postinfarction phase in patients on placebo as well as in patients 12-24 h after captopril intake. Plasma atrial natriuretic peptide concentration (ANP) was increased immediately after myocardial infarction, but ANP levels almost normalized in patients with captopril treatment, while they continued to be elevated in patients on placebo. The only technical parameter able to predict left ventricular volume increases was the sphericity index (28.7 vs. 35.7; p = 0.07). We concluded that morphologic deformation and filling pressures as estimated from elevated ANP levels are major factors promoting remodelling following myocardial infarction. ACE inhibitors might exert their favorable effect predominantly by reducing filling pressure.     

Role of p38alpha MAPK in cardiac apoptosis and remodeling after myocardial infarction

Acute coronary occlusion results in ischemia-mediated death of cardiomyocytes. In the days and weeks following myocardial infarction (MI), left ventricular remodeling occurs that is characterized by persistent cardiomyocyte apoptosis, thinning and fibrosis at the site of infarction, ventricular chamber dilatation, and growth of remaining viable cardiomyocytes. The p38 mitogen-activated protein kinase (MAPK) signaling cascade has been implicated in the remodeling process. In this work, mice with cardiac-specific expression of a dominant negative mutant form of p38 MAPK (DN-p38alpha) were subjected to MI by occlusion of the left coronary artery. Acute ischemia area was determined by transthoracic echocardiography 2 h after MI surgery, and was found to be nearly identical in DN-p38 mice and their wild-type littermates. Seven days after MI, mice were subjected to repeat echocardiography and histological examination of infarct size. DN-p38 mice had markedly reduced infarct size and increased ventricular systolic function 7 days after MI when compared to wild-type littermates. In addition, DN-p38 mice had less cardiomyocyte apoptosis than wild-type mice in the infarct border zone. Recently, it was discovered that Bcl-X(L) deamidation occurs in vivo, and this results in Bcl-X(L) degradation that sensitizes cells to apoptosis by enhancing BAX activity. Bcl-X(L) deamidation was found to occur in the cardiac tissue of wild-type mice after MI, but was reduced in DN-p38 mice. These results establish that p38 MAPK activity is required for pathological remodeling after MI and suggest that p38 MAPK may promote cardiomyocyte apoptosis through Bcl-X(L) deamidation.     

低声压级水平次声对心肌梗死大鼠心肌纤维化的影响及机制研究

目的:观察次声对心肌梗死后大鼠心肌纤维化的影响及相关机制研究。方法:将30只SD大鼠随机分为3组,即假手术组、心肌梗死组及次声治疗组,每组10只。采用低声压级水平次声对心肌梗死大鼠治疗1周(2次/d,0.5 h/次),观察心肌胶原和血浆NO和血管紧张素-Ⅱ(AngⅡ)及大鼠心功能的变化。结果:与心肌梗死组大鼠比较(未治疗组),次声治疗组大鼠心肌胶原表达明显减少,血浆NO含量显著增加(P0.01),血浆AngⅡ含量显著降低(P0.01),心功能明显改善(P0.01)。结论:低声压级水平次声治疗能够明显减轻心肌梗死后大鼠心肌纤维化,其作用可能与血浆NO及AngⅡ含量变化有关。