扩张型心肌病患者恶性室性心律失常与心率变异性关系分析

目的:了解扩张型心肌病患者恶性心律失常(MVA)与心率变异性(heart rate variability,HRV)的关系,探讨扩心病患者体内自主神经变化的临床意义。方法:选择扩心病患者48例作为研究对象,同时按照年龄配对,取48例正常者作为对照组,对其行24小时动态心电图检查,依据其是否出现恶性心律失常分为恶性室性心律失常(MVA+)组及单纯扩张型心肌病(MVA-)组,分析组间HRV的差异。结果:与对照组比较,单纯扩张型心肌病(MVA-)组HRV时域指标(SDNN、SDANN、RMSSD)均有降低(P<0.05)L与(MVA-)组相比,恶性室性心律失常(MVA+)组HRV相关指标进一步降低(P<0.05)。结论:自主神经功能异常是扩张型心肌病患者恶性心律失常的重要危险因子,可能可以用HRV预测其发生恶性心律失常危险性。     

Danon disease: a focus on processing of the novel LAMP2 mutation and comments on the beneficial use of peripheral white blood cells in the diagnosis of LAMP2 deficiency

Danon disease (DD) is a monogenic X-linked disorder characterized by cardiomyopathy, skeletal myopathy and variable degrees of intellectual disability. DD develops due to mutations in the gene encoding lysosomal-associated membrane protein 2 (LAMP2). We report on a family exhibiting the clinical phenotype comprising of hypertrophic cardiomyopathy and ventricular pre-excitation, myopia and mild myopathy in two male patients and cardiomyopathy and myopia in a female patient. The diagnosis of DD in this family was based on the assessment of the clinical phenotypes and the absence of LAMP2 in skeletal and/or cardiac muscle biopsy specimens. Sequence analysis of the LAMP2 gene and its mRNA revealed a novel LAMP2 mutation (c.940delG) in all three patients. Approximately 25% of the female patient's cardiomyocytes were LAMP2 positive apparently due to the unfavorable skewing of X chromosome inactivation. We further performed qualitative LAMP2 immunohistochemistry on peripheral white blood cells using the smear technique and revealed the absence of LAMP2 in the male patients. LAMP2 expression was further assessed in granulocytes, CD4+ and CD8+ T lymphocytes, CD20+ B lymphocytes, CD14+ monocytes and CD56+ natural killer cells by quantitative polychromatic flow cytometry. Whereas the male DD patients lacked LAMP2 in all WBC populations, the female patient expressed LAMP2 in 15.1% and 12.8% of monocytes and granulocytes, respectively. LAMP2 expression ratiometrics of highly vs. weakly expressing WBC populations discriminated the DD patients from the healthy controls. WBCs are thus suitable for initial LAMP2 expression testing when DD is a differential diagnostic option. Moreover, flow cytometry represents a quantitative method to assess the skewing of LAMP2 expression in female heterozygotes. Because LAMP2 is a major protein constituent of the membranes of a number of lysosome-related organelles, we also tested the exocytic capacity of the lytic granules from CD8+ T lymphocytes in the patient samples. The degranulation triggered by a specific stimulus (anti-CD3 antibody) was normal. Therefore, this process can be considered LAMP2 independent in human T cells. The c.940delG mutation results in a putatively truncated protein (p.A314QfsX32), which lacks the transmembrane domain and the cytosolic tail of the wild-type LAMP2. We tested whether this variant becomes exocytosed because of a failure in targeting to late endosomes/lysosomes. Western blotting of cardiac muscle, WBCs and cultured skin fibroblasts (and their culture media) showed no intra- or extracellular truncated LAMP2. By comparing the expression pattern and intracellular targeting in cultured skin fibroblasts of normal LAMP2 isoforms (A, B and C) tagged with green fluorescent protein (GFP) and the A314Qfs32-GFP fusion, we found that the A314Qfs32-GFP protein is not even expressed. These observations suggest that the truncated protein is unstable and is co-translationally or early post-translationally degraded.     

Diacylglycerol acyl transferase 1 overexpression detoxifies cardiac lipids in PPARγ transgenic mice

Accumulation of excess lipids is associated with heart failure. The effects of transgenic expression of diacylglycerol acyl transferase 1 (DGAT1) in cardiomyocytes is controversial. We explored whether mice expressing DGAT1 via the myosin heavy chain (MHC) promoter develop heart dysfunction with aging or after crossing with mice over expressing peroxisome proliferator-activated receptor γ (PPARγ) in the heart. MHC-DGAT1 transgenic mice had increased heart triglyceride but no evidence of heart dysfunction, even up to age 12 months. The MHC-DGAT1 transgene improved heart dysfunction and survival of MHC-PPARγ-expressing transgenic mice. Both diacylglycerol and ceramide levels in the heart were reduced by this cross, as were the levels of several mRNAs of genes involved in lipid metabolism. There were fewer large lipid droplets in MHC-DGAT1×MHC-PPARγ mice compared with MHC-PPARγ, but total lipid content was not changed. Therefore, overexpression of DGAT1 is not toxic to the heart but reduces levels of toxic lipids and improves lipotoxic cardiomyopathy. Moreover, the beneficial effects of DGAT1 illustrate the interrelationship of several lipid metabolic pathways and the difficulty of assigning benefit to an isolated change in one potentially toxic lipid species.     

地高辛联合左西孟旦治疗扩张型心肌病的疗效及对血清HMGB1、sICAM-1、sST2水平的影响

目的:探讨地高辛联合左西孟旦治疗扩张型心肌病的疗效及对血清高迁移率族蛋白B1(HMGB1)、溶性细胞间粘附分子-1(s ICAM-1)、可溶性ST2蛋白(s ST2)水平的影响。方法：选择2016年2月至2018年2月我院接诊的90例扩张型心肌病患者作为本研究对象,依照随机数表法分为观察组44例和对照组46例,对照组在常规治疗基础上给予左西孟旦治疗,观察组在对照组基础上联合地高辛治疗,两组均连续治疗4周。比较两组的临床疗效、左室射血分数(LVEF)、左室舒张末内径(LVEDD)、血清氨基末端B型钠尿肽前体(NT-proBNP)、HMGB1、s ICAM-1、s ST2水平的变化及不良反应的发生情况。结果:治疗后,观察组临床疗效总有效率为93.18%(41/44),明显高于对照组(76.09%,P0.05);观察组LVEF明显高于对照组,LVEDD、血清NT-proBNP水平均明显低于对照组[(46.50±5.21)%vs.(41.20±4.12)%,(54.94±2.29)mm vs.(59.30±2.38)mm,(494.31±75.95)ng/L vs.(589.56±89.40)ng/L](P0.05);观察组血清HMGB1、s ICAM-1、s ST2水平均明显低于对照组低[(8.42±1.23)pg/mL vs.(13.76±1.70)pg/mL,(122.93±11.03)μg/L vs.(141.58±13.04)μg/L,(0.08±0.02)ng/mL vs.(0.15±0.03)ng/mL](P0.05)。治疗期间,两组不良反应发生率分别为9.09%和6.52%,组间差异无统计学意义(P0.05)。结论:地高辛联合左西孟旦治疗扩张型心肌病患者的效果显著优于单用左西孟旦治疗,其可更有效降低患者血清HMGB1、s ICAM-1、s ST2的表达,改善患者心功能,且安全性高。     

解酒护肝饮解酒及对急慢酒精性肝损伤的保护作用

为了评价解酒护肝饮解酒效果及其对急、慢性酒精性肝损伤保护作用机制,本研究通过建立醉酒模型,确定致醉剂量;通过醉酒睡眠实验比较解酒护肝饮解酒特性;通过测定醉酒小鼠血乙醇含量的变化,研究解酒护肝饮对乙醇代谢的影响;通过建立急慢性酒精性肝损伤模型,测定AST、ALT、SOD活性,GSH、MDA水平,HE染色切片观察肝组织形态学的变化。研究发现小鼠最佳致醉剂量为11 m L/kg;与模型组比较,解酒护肝饮高(HD)、中剂量组(MD)均可延长醉酒时间、缩短醒酒时间(p<0.05),高、中剂量组可降低酒精灌胃后2 h、3 h时间点血乙醇含量(p<0.05);与模型组比较,急慢性酒精肝损伤模型各剂量组均能显著降低血清AST、ALT活性(p<0.05),急性酒精性肝损伤模型中,各剂量组肝组织SOD、GSH水平上升(p<0.05),MDA水平下降(p<0.05),而在慢性酒精性肝损伤模型肝组织中,低剂量组(LD)的SOD、GSH及MDA水平没有统计学差异;病理切片观察可见,急慢性酒精肝损伤模型高、中、低剂量组均能显著改善肝组织因乙醇而导致的肝损伤,并且高、中剂量组效果较好。本研究表明解酒护肝饮可显著延长醉酒时间,缩短醒酒时间,降低血乙醇的含量,对酒精诱导的肝损伤有较好的保护作用。     

Eliciting α7‐nAChR exerts cardioprotective effects on ischemic cardiomyopathy via activation of AMPK signalling

Our previous studies have reported that agonist of α7 nicotinic acetylcholine receptors prevented electrophysiological dysfunction of rats with ischaemic cardiomyopathy (ICM) by eliciting the cholinergic anti‐inflammatory pathway (CAP). Adenosine monophosphate‐activated protein kinase (AMPK) signalling is widely recognized exerting cardioprotective effect in various cardiomyopathy. Here, we aimed to investigate whether the protective effects of the CAP are associated with AMPK signalling in ICM. In vivo, coronary artery of rats was ligated for 4 weeks to induce the ICM and then treated with PNU‐282987 (CAP agonist) and BML‐275 dihydrochloride (AMPK antagonist) for 4 weeks. In vitro, primary macrophages harvested from rats were induced inflammation by Lipopolysaccharide (LPS) treatment and then treated with PNU‐282987 and BML‐275 dihydrochloride. In vivo, exciting CAP by PUN‐282987 elicited an activation of AMPK signalling, alleviated ventricular remodeling, modified the cardiac electrophysiological function, reduced the cardiac expression of collagens and inflammatory cytokines and maintained the integrity of ultrastructure in the ischemic heart. However, the benefits of CAP excitation were blunted by AMPK signaling antagonization. In vitro, excitation of the CAP was observed inhibiting the nuclear transfer of NF‐κB p65 of macrophages and promoting the transformation of Ly‐6C^high macrophages into Ly‐6C^low macrophages. However, inhibiting AMPK signalling by BML‐275 dihydrochloride reversed the CAP effect on LPS‐treated macrophages. Finally, our findings suggest that eliciting the CAP modulates the inflammatory response in ICM through regulating AMPK signalling.     

Protein kinase C in the human heart: differential regulation of the isoforms in aortic stenosis or dilated cardiomyopathy

Objectives Protein kinase C (PKC) is a central enzyme in the regulation of growth and hypertrophy. Little was known on PKC isoform regulation in human heart. Goal of this study was to characterize the isoforms of protein kinase C in human heart, their changes during ontogenesis, and their regulation in myocardial hypertrophy and heart failure. Methods In left ventricular and atrial samples from adults with end-stage dilated cardiomyopathy (DCM), from adults with severe aortic stenosis (AS), from small infants undergoing repair of ventricular septal defects, and from healthy organ donors (CO), activity of protein kinase C and the expression of its isozymes were examined. Results In the adult human heart, the isoforms PKC-α, PCK-β, PKC-δ, PKC-ε, PKC-λ/-ι, and PKC-ζ were detected both on protein and on mRNA level. All isozymes are subjected to downregulation during ontogenesis. No evidence, however, exists for an isoform shift from infancy to adulthood. DCM leads to a pronounced upregulation of PKC-β. Severe left ventricular hypertrophy in AS, however, recruits a distinct isoform pattern, i.e., isoforms PKC-α, PKC-δ, PKC-ε, PKC-λ/-ι, and PKC-ζ are upregulated, whereas PKC-β is not changed under this condition. Conclusion This work gives evidence for a differential recruitment of human PKC isoforms in various forms of myocardial hypertrophy and heart failure. Gregor Simonis and Steffen K. Briem contributed equally to this work.     

伊伐布雷定联合美托洛尔对缺血性心肌病PCI术患者心功能、预后及血清炎性因子的影响

目的:探讨伊伐布雷定联合美托洛尔对缺血性心肌病(ICM)行经皮冠状动脉介入术(PCI)术后患者心功能、预后及血清炎性因子的影响。方法:选取2018年7月-2019年4月我院收治的行PCI术的ICM患者98例,将随机数字表法分为实验组与对照组,各49例,对照组给予美托洛尔治疗,实验组给予伊伐布雷定联合美托洛尔治疗,对比两组优良率、心功能、预后、不良反应及血清炎性因子。结果:实验组治疗8周后的优良率为93.75%(45/48),高于对照组的70.83%(34/48)(P<0.05)。实验组治疗8周后左心室射血分数(LVEF)、6 min步行试验(6MWT)距离高于对照组,N末端B型钠尿肽原(NT-proBNP)、24 h心率(24hHR)低于对照组(P<0.05)。实验组治疗8周后肿瘤坏死因子-α(TNF-α)、超敏C反应蛋白(hs-CRP)、白介素-6(IL-6)低于对照组(P<0.05)。实验组3个月内再住院率、3个月内病死率低于对照组(P<0.05)。两组不良反应发生率比较无差异(P>0.05)。结论:相较于单用美托洛尔,联合伊伐布雷定治疗PCI术后的ICM患者,可更好地改善患者心功能,降低炎性因子水平,且安全性较好,改善患者预后的效果更佳。     

Endothelin 1 gene as a modifier in dilated cardiomyopathy

Dilated cardiomyopathy (DCM) is a myocardial disease of unknown etiology with left ventricular dilatation and impaired myocardial contractility leading to heart failure. It is considered to be a multifactorial disorder with the interplay of both genetic and environmental factors. One of the possible genes implicated in DCM is endothelin 1 (EDN1). The genetic variants of EDN1 may be involved in the pathophysiology of DCM hence the entire EDN1 gene was screened to examine for the possible genotypic associations with DCM. A total of 115 DCM patients and 250 control subjects were included in the present study. PCR based SSCP analysis was carried out followed by commercial sequencing. Screening of EDN1 revealed two common and two rare polymorphisms. Allelic and genotypic frequencies were estimated in patient and control groups by appropriate statistical tests. The heterozygotes of insertion variation (+ 138A) were found to exhibit four-fold increase risk to DCM (OR = 4.12, 95% CI 2.10–8.08; p = 0.0001). The two rare variants (G>A transition (rs150035515) at c.90 and C>T transition (rs149399492) at c.119) observed in the present study were found to be unique in DCM. The secondary mRNA structures of these variations were found to have less free energy than wild type. The haplotype analysis revealed 4A–T to be risk haplotype for DCM (OR 5.90, 95% CI 2.29–15.25, p = 0.0001). In conclusion, EDN1 polymorphisms (+ 138A, A30A, T40I) appear to play a significant role in the pathogenesis of DCM, as they influence the stability of protein. The increased EDN1 production may lead to constriction of coronary arteries, reducing coronary blood flow which may in turn increase the load on left ventricle, impairing contractility of the heart resulting in a DCM phenotype, an end stage of heart failure.