促红细胞生成素与慢性心衰心功能的关系

探讨慢性心衰患者不同心功能阶段内源性促红细胞生成素（EPO）水平的变化及EPO与心功能、EPO与BNP、hs-CRP、TNF-α之间的关系,从而寻找慢性心衰早期诊断的新生物标志物。选取2013年4—10月入住我院心内科及干疗科的临床诊断为慢性心衰的患者74例为观察组,以同期住院的30例无心血管病的人群为对照组,测定相关指标。观察组EPO水平明显升高 (P＜0.01);观察组EPO水平随心功能分级（NYHA）增加而显著升高,各心功能分级间有显著性差异(P＜0.01);经Pearson相关性分析观察组EPO与BNP、hs-CRP、TNF-α呈正相关关系,与LEVF呈负相关关系;经Pearson相关性分析观察组BNP与TNF-α呈正相关,与hs-CRP呈正相关。血清EPO水平在慢性心衰患者中明显升高,且随心衰的加重而明显增加;慢性心衰患者EPO随BNP、TNF-α、hs-CRP水平升高以及LVEF的降低而升高;血清EPO水平与心衰严重程度密切相关,可作为心衰病情评估的一个指标,检测慢性心衰患者EPO水平对心衰患者的诊治具有重要意义。     

Erythropoietin inhibits angiotensin Ⅱ induced cardiomyocyte hypertrophy in vitro via activating PI3K/Akt-eNOS pathway

Objective To explore the effect of erythropoietin (EPO) on angiotensin Ⅱ (Ang Ⅱ) induced neonatal rat cardiomyocyte hypertrophy and the association with PI3K/Akt-eNOS signaling pathway. Methods Cardiomyocytes were isolated from new-born Sprague-Dawley rats and stimulated by Ang Ⅱ in vitro. The cell surface area and mRNA expression of atrial natriuretic factor (ANF) of cardiomyocytes were determined in the presence and absence of various concentrations of EPO, phosphatidylinositol 3'-kinase (PI3K) inhibitor LY294002 and nitric oxide synthase (NOS) inhibitor L-NAME. Intracellular signal molecules, such as Akt, phosphorylated Akt, eNOS and phosphorylated eNOS protein expressions were detected by western blot. Nitric oxide (NO) level in the supernatant of cultured cardiomyocytes was assayed by NO assay kit. Results EPO (20 U/ml) significantly inhibited Ang Ⅱ induced cardiomyocyte hypertrophy as shown by decreased cell surface area and ANF mRNA expression (all P ＜0.05). EPO also activated Akt and enhanced the expression of eNOS and its phosphorylation (all P ＜ 0.05), increased the NO production (P ＜0.01). These effects could be partially abolished by cotreatment with LY294002 or L-NAME (all P ＜ 0.05). Conclusion EPO attenuates Ang Ⅱ induced cardiomyocytes hypertrophy via activating PI3K-Akt-eNOS pathway and promoting NO production.     

Erythropoietin inhibits angiotensin Ⅱ induced cardiomyocyte hypertrophy in vitro via activating PI3K/Akt-eNOS pathway

Objective To explore the effect of erythropoietin (EPO) on angiotensin Ⅱ (Ang Ⅱ) induced neonatal rat cardiomyocyte hypertrophy and the association with PI3K/Akt-eNOS signaling pathway. Methods Cardiomyocytes were isolated from new-born Sprague-Dawley rats and stimulated by Ang Ⅱ in vitro. The cell surface area and mRNA expression of atrial natriuretic factor (ANF) of cardiomyocytes were determined in the presence and absence of various concentrations of EPO, phosphatidylinositol 3'-kinase (PI3K) inhibitor LY294002 and nitric oxide synthase (NOS) inhibitor L-NAME. Intracellular signal molecules, such as Akt, phosphorylated Akt, eNOS and phosphorylated eNOS protein expressions were detected by western blot. Nitric oxide (NO) level in the supernatant of cultured cardiomyocytes was assayed by NO assay kit. Results EPO (20 U/ml) significantly inhibited Ang Ⅱ induced cardiomyocyte hypertrophy as shown by decreased cell surface area and ANF mRNA expression (all P ＜0.05). EPO also activated Akt and enhanced the expression of eNOS and its phosphorylation (all P ＜ 0.05), increased the NO production (P ＜0.01). These effects could be partially abolished by cotreatment with LY294002 or L-NAME (all P ＜ 0.05). Conclusion EPO attenuates Ang Ⅱ induced cardiomyocytes hypertrophy via activating PI3K-Akt-eNOS pathway and promoting NO production.     

心房颤动导管消融围手术期抗凝诱发再次心包压塞一例

患者男,65岁。心房颤动导管消融术中发生心包压塞成功救治后,围手术期抗凝治疗诱发再次心包压塞。提示消融能量致心房壁爆裂的患者,围手术期早期抗凝治疗应慎重;抗凝强度应较正常偏低。同时延长心包引流管留置时间;密切观察患者局部及全身情况,及时做出相应处理。     

冠状动脉瘤样扩张病因与治疗的认识

冠状动脉瘤样扩张（coronary artery ectasia,CAE)是冠状动脉粥样硬化的一种疾病,最早报道于1983年[1-2];有相关文献报道冠状动脉瘤样扩张是冠状动脉梗阻型疾病的变异性表现,而不是特有疾病。冠状动脉瘤样扩张是心血管疾病危险因素之一,因扩张段产生缓慢血流量,能诱发心绞痛及心肌梗死。随着冠状动脉造影、冠状动脉CT、磁共振血管造影及冠脉介入治疗的普及,冠状动脉瘤样扩张的有关报道及研究越来越多;但是确切病因仍不清楚,本文对CAE的病因、治疗及预后作一综述。     

Erythropoietin inhibits angiotensin Ⅱ induced cardiomyocyte hypertrophy in vitro via activating PI3K/Akt-eNOS pathway

Objective To explore the effect of erythropoietin (EPO) on angiotensin Ⅱ (Ang Ⅱ) induced neonatal rat cardiomyocyte hypertrophy and the association with PI3K/Akt-eNOS signaling pathway. Methods Cardiomyocytes were isolated from new-born Sprague-Dawley rats and stimulated by Ang Ⅱ in vitro. The cell surface area and mRNA expression of atrial natriuretic factor (ANF) of cardiomyocytes were determined in the presence and absence of various concentrations of EPO, phosphatidylinositol 3'-kinase (PI3K) inhibitor LY294002 and nitric oxide synthase (NOS) inhibitor L-NAME. Intracellular signal molecules, such as Akt, phosphorylated Akt, eNOS and phosphorylated eNOS protein expressions were detected by western blot. Nitric oxide (NO) level in the supernatant of cultured cardiomyocytes was assayed by NO assay kit. Results EPO (20 U/ml) significantly inhibited Ang Ⅱ induced cardiomyocyte hypertrophy as shown by decreased cell surface area and ANF mRNA expression (all P ＜0.05). EPO also activated Akt and enhanced the expression of eNOS and its phosphorylation (all P ＜ 0.05), increased the NO production (P ＜0.01). These effects could be partially abolished by cotreatment with LY294002 or L-NAME (all P ＜ 0.05). Conclusion EPO attenuates Ang Ⅱ induced cardiomyocytes hypertrophy via activating PI3K-Akt-eNOS pathway and promoting NO production.     

体外培养猪骨髓间充质干细胞hHCN2基因的转染及表达

背景：通过超极化活化的阳离子电流调制心脏自律性成为该领域研究的焦点。 目的：将起搏基因质粒CMV-hHCN2-3xha-IRES-EGFP转染到猪骨髓间充质干细胞,检测其在骨髓间充质干细胞中核酸和蛋白水平是否表达。 方法：采用单纯直接贴壁法或密度梯度离心结合直接贴壁法分离、纯化、增殖获得骨髓间充质干细胞,脂质体2000转染质粒CMV-hHCN2-3xha-IRES-EGFP至骨髓间充质干细胞。 结果与结论：单纯直接贴壁法收集到的细胞增殖速度慢,而密度梯度离心结合直接贴壁法细胞增殖速度较快,原代细胞培养第3代后转染48 h荧光显微镜下可见骨髓间充质干细胞发出荧光,对照组无荧光。细胞转染率达15%-20%。RT-PCR检测及Western blot 印迹检测证明了hHCN2基因在骨髓间充质干细胞有表达。密度梯度离心结合直接贴壁法较单纯直接贴壁法培养骨髓间充质干细胞的培养效率更高。转染目的基因hHCN2的骨髓间充质干细胞蛋白有表达。     

促红细胞生成素抑制急性梗死心肌炎症因子的表达

背景：研究表明,炎症细胞因子可影响心肌梗死后的预后,在心脏重构的进程中起重要作用。同时促红细胞生成素的促红细胞生成之外的非造血效应也被广泛证实：促红细胞生成素可通过与靶细胞膜表面的促红细胞生成素受体结合而减少炎症反应,从而减少心肌缺血后的再灌注损伤。 目的：观察重组人促红细胞生成素对大鼠急性心肌梗死后心脏重构中炎症因子表达的影响。 方法：通过结扎左冠状动脉前降支建立SD大鼠急性心肌梗死模型,分为5组,假手术组注射生理盐水,手术对照组造模后注射生理盐水,SB203580组造模后注射p38 MAPK的高选择性阻断剂SB203580,促红细胞生成素组造模后注射促红细胞生成素注射液,促红细胞生成素+SB203580组造模后注射促红细胞生成素+ SB203580混合液,分别在造模前、造模后1 d、1周、2周及4周进行尾静脉采血,酶联免疫吸附法检测血清中白细胞介素1β、白细胞介素6、肿瘤坏死因子α水平变化。 结果与结论：造模前各组大鼠血清白细胞介素1β、白细胞介素6、肿瘤坏死因子α检测值差异均无显著性意义(P > 0.05)。假手术组各时段白细胞介素1β、白细胞介素6、肿瘤坏死因子α检测值差异无显著性意义(P > 0.05),其余4组不同时段各因子检测值呈现造模后1 d最高,造模后4周回降的趋势(P < 0.05)。造模后手术对照组血清各因子检测值较其他组升高明显,而假手术组血清各因子检测值均低于其他4组(P < 0.05);使用药物进行干预的3组中,促红细胞生成素+SB203580组各因子检测值较低(P < 0.05),而促红细胞生成素组与SB203580组各因子检测值差异无显著性意义(P > 0.05)。提示重组人促红细胞生成素抑制了大鼠急性心肌梗死后心脏重构中炎症因子白细胞介素1β、白细胞介素6、肿瘤坏死因子α的表达,重组人促红细胞生成素抑制炎症因子表达的机制可能与转化生长因子β1-TAK1-p38 MAPK信号路径相关。     

超极化激活环化核苷酸门控通道2基因转染后293T细胞中目的基因的表达

背景：研究证实超极化激活环化核苷酸门控通道电流在调控心脏的自发搏动中起着非常重要的作用。 目的：观察超极化激活环化核苷酸门控通道2基因重组质粒转染后293T细胞中目的基因在核酸和蛋白质水平的表达。 方法：采用脂质体转染试剂Lipofectamine2000将含全长超极化激活环化核苷酸门控通道2基因的质粒CMV-hHCN2-3xHA-IRES-EGFP转染至293T细胞中。 结果与结论：用脂质体转染试剂Lipofectamine2000成功地将质粒CMV-hHCN2-3xHA-IRES-EGFP转染至293T细胞中。反转录PCR定性地检测到在100-250 bp处可见高度特异DNA的条带,与携带超极化激活环化核苷酸门控通道2基因的质粒扩增出的片段位置相同。经过RT-PCR、Western-blot方法检测提示超极化激活环化核苷酸门控通道2基因在mRNA和蛋白水平都有高表达。表明质粒CMV-hHCN2-3xHA-IRES-EGFP成功转染后的293T细胞中超极化激活环化核苷酸门控通道2基因可以在核酸和蛋白水平表达。     

促红细胞生成素对新生大鼠心脏成纤维细胞氧化应激时一氧化氮合酶系统的影响

目的:探讨促红细胞生成素(EPO)对心脏成纤维细胞(CFs)氧化应激时一氧化氮合酶(NOS)系统的影响,以及PI3-K/Akt信号途径在其中的作用。方法:应用胰酶和胶原酶双酶法分离培养新生大鼠CFs,EPO、过氧化氢(H2O2)和PI3-K抑制剂LY294002不同因素干预。化学酶法检测CFs培养液中的NO浓度以及总NOS和其亚型的活性。Western blot检测Akt、p-Akt、eNOS、iNOS蛋白的表达。结果:氧化应激使CFs中iNOS的表达显著上调,活性增强;eNOS蛋白的表达明显下降。同时CFs培养液中NO的合成增加显著,达到(25.94±2.57)μmol/L,是对照组的4倍多(P<0.01)。EPO显著抑制CFs氧化应激时iNOS蛋白的表达,同时上调eNOS蛋白,总的NO浓度也显著下降。Akt磷酸化形式p-Akt的表达水平也明显提高。PI3-K抑制剂LY294002能阻断EPO促进eNOS和p-Akt蛋白表达的作用,但EPO抑制iNOS的作用不能完全被阻断。结论:EPO能促进新生大鼠CFs氧化应激时eNOS蛋白的合成,抑制iNOS蛋白的表达。其促进eNOS的表达是通过激活PI3-K/Akt细胞信号途径来实现,抑制iNOS的机制有待进一步探讨。