目的 讨论研究靶向磷酸二酯酶5(PDE5)的短发夹RNA(shRNA)导入后的间充质干细胞(MSCs)对心肌梗死后心功能影响价值.方法 选择2021年3月-7月实验室的20只体重为280-300 g的雄性SD大鼠,运用随机分配的方式设置为假手术组、对照组、MSCs组、shPDE5组、MSCs shPDE5组,对假手术组进行穿线但是不结扎左冠状动脉前降支操作;对对照组进行穿线并结扎前降支操作;对MSCs组进行MSCs细胞注射操作;对shPDE5组进行shPDE5细胞注射操作;对MSCs shPDE5组进行MSCs shPDE5细胞注射操作.对比五组大鼠经过实验后的心肌梗死后心功能变化情况、治疗有效率;对比MSCs组、shPDE5组、MSCs shPDE5组的移植细胞存活率.结果 对照组的左室射血分数明显低于假手术组、MSCs组、shPDE5组、MSCs shPDE5组(P<0.05),但MSCs shPDE5组与假手术组无显著差异(P>0.05),分别高于MSCs组与shPDE5组(P<0.05).MSCs shPDE5组的移植细胞存活率分别高于MSCs组、shPDE5组(P<0.05).MSCs shPDE5组细胞移植方式各组整体的治疗效果水平明显高于手术方式各组(P<0.05).结论 将MSCs shPDE5细胞移植方式运用到心肌梗死治疗过程中,可以有效改善大鼠心功能,而且心功能改善效果水平明显超过shPDE5细胞移植、MSCs细胞移植、手术等治疗方式的改善效果水平,而且运用细胞移植方式可以有效提升整体的治疗效果水平.

Dysregulation of long non-coding RNA (IncRNA) antisense non-coding RNA in the INK4 locus (ANRIL) is associated with the risk of myocardial infarction (MI). Therefore, the present study aimed to determine the mechanisms underlying this association, which is currently poorly understood, to the best of our knowledge. The current study used an in vitro myocardial ischemia and reperfusion (MI/R) model, in which H9c2 cardiomyocytes were exposed to hypoxia/reoxygenation (H/R), which demonstrated that ANRIL expression was downregulated and that ANRIL positively regulated sirtuin 1 (SIRT1) expression following H/R injury. Subsequently, it was demonstrated that ANRIL upregulated SIRT1 expression by sponging microRNA-181a (miR-181a). In addition, ANRIL overexpression reduced lactate dehydrogenase release and apoptosis of H9c2 cardiomyocytes exposed to H/R, indicating that ANRIL prevented H/R-induced cardiomyocyte injury. Moreover, both miR-181a overexpression and SIRT1 knockdown significantly decreased the protective effects of ANRIL on H/R-induced cardiomyocyte injury, thus demonstrating that SIRT1 upregulation via sponging miR-181a is a critical mechanism that mediates the function of ANRIL. These results provided a novel mechanistic insight into the role of ANRIL in H/R-injured cardiomyocytes and suggested that the ANRIL/miR-181a/SIRT1 axis may be a therapeutic target for reducing MI/R injury.

目的 探讨磷酸二酯酶5(PDE5)短发夹RNA(shRNA)重组腺病毒载体[PDE5shRNA]导入后的间充质干细胞(MSCs)对心肌梗死(MI)后心室重构的影响.方法 构建PDE5shRNA转染的MSCs,结扎大鼠冠状动脉左前降支(LAD)建立MI模型,随机分成假手术组(n =4,只穿线不结扎LAD);模型组(n =4,穿线并结扎LAD建立MI模型);MSCs-Ad-Null组(n =4,MI模型基础上注射经空白腺病毒载体转染的MSCs);MSCs-shRNA-PDE5组(n =4,MI模型基础上注射经PDE5shRNA转染的MSCs).建模后,每组各取1只大鼠取心脏进行病理检验,用于判断造模成功,余大鼠入组观察相关指标.4周后心脏超声检测各组大鼠心功能指标[左室收缩末期内径(LVESd)、左室舒张末期内径(LVEDd)、左室射血分数(LVEF)和左室短轴缩短率(FS)].HE染色观察心肌病理改变,Masson染色观察心肌纤维化情况,qRT-PCR方法检测大鼠心肌中PDE5、环磷酸鸟苷(cGMP)和蛋白激酶(PKG)mRNA的相对表达量.结果 实验4周后,(1)心脏超声示:MI各组大鼠心室腔明显扩张,左心室心肌收缩功能明显降低.与假手术组相比,MI各组大鼠的LVESd、LVEDd增大,LVEF、FS降低(P均＜0.05);与模型组相比,MSCs-Ad-Null组、MSCs-shRNA-PDE5组的 LVESd、LVEDd 降低,MSCs-shRNA-PDE5组的 LVEF、FS 升高(P均＜0.05).(2)心肌病理结果示:与模型组比较,MSCs-Ad-Null组、MSCs-shRNA-PDE5组的心肌病理改变逐渐减轻.(3)qRT-PCR法结果示:与假手术组相比,模型组、MSCs-Ad-Null 组PDE5 mRNA表达升高、cGMP和PKG mRNA表达降低(P均＜0.05);按模型组→MSCs-Ad-Null 组→MMSCs-shRNA-PDE5 组之序,PDE5 mRNA 表达递降,cGMP、PKG mRNA 表达递升(P＜0.05,P＜0.01).结论 PDE5shRNA导入后的MSCs可改善MI后心功能,减轻心肌病理改变及心肌纤维化,同时通过调节PDE5、cGMP、PKG的表达,发挥抑制心室重构的作用.

目的 通过短发夹RNA(shRNA)延长对磷酸二酯酶5(PDE5)的抑制作用,探索shRNA-PDE5对细胞存活率的影响以及对心肌梗死后心功能及心室重构的作用及其机制.方法 (1)细胞实验:分离SD大鼠骨髓间充质干细胞(MSCs),将Ad-shRNA-PDE5和Ad-Null分别感染体外培养的MSCs,嘌呤霉素筛选稳定表达它们的细胞株,分为shPDE5组和空白对照载体(Ad-Null)组,另设正常组.缺口末端标记法染色检测各组缺氧缺糖(OGD)诱导的稳转MSCs细胞凋亡情况.(2)动物实验:通过结扎左冠状动脉前降支方法建立雄性SD大鼠心肌梗死模型.按照体重将大鼠随机分成6组:假手术组、模型组、对照组、第1转染组、第2转染组和第3转染组,在结扎前降支后,立即在心肌梗死区及周边区4个点各注射经不同处理的MSCs细胞.用蛋白质印迹法检测大鼠心肌中转化生长因子-β(TGF-β)和成纤维细胞生长因子2(FGF2)的蛋白表达水平(光密度值);Masson染色检测心肌梗死区及周边区组织的纤维化情况.结果 (1)shPDE5组、Ad-Null组和正常组的凋亡细胞数分别为(6.00±1.00),(14.33±1.52)和(5.33±1.52)个,shPDE5组与Ad-Null组比较,凋亡细胞数明显减少,差异有统计学意义(P<0.05).(2)假手术组、模型组、对照组、第1转染组、第2转染组和第3转染组的FGF2蛋白表达水平分别为0.11±0.02,0.22±0.05,0.27±0.08,0.28±0.04,0.33±0.05和0.50±0.03;这6组的TGF-β蛋白表达水平分别为0.18±0.02,0.91±0.03,0.72±0.01,0.72±0.03,0.48±0.02和0.27±0.09.第2转染组、第3转染组与假手术组、模型组、对照组和第1转染组比较,上述指标的差异均有统计学意义(均P<0.05).对照组、第1转染组、第2转染组和第3转染组与模型组比较,大鼠梗死周边区心肌组织细胞纤维化程度依次减弱.结论 shRNA -PDE5 可通过减少细胞凋亡,调节相关因子的表达,抑制心肌细胞纤维化,从而改善心功能.

目的 本文旨在研究对ST段抬高型心肌梗死(ST segment elevation myocardial infarction,STEMI)患者经皮冠状动脉介入治疗(percutaneous coronary intervention,PCI)术中发生无复流情况后运用尼可地尔相对常规硝酸甘油注射的效果比较.方法 回顾性选取2019年1—12月于我院心血管内科住院治疗的冠心病需行PCI术的患者共计84例.根据PCI术中无复流现象处理方式不同分为观察组41例及对照组43例.观察组无复流后注射尼可地尔,对照组注射硝酸甘油.分别对两组患者的用药后无复流发生率、治疗后ST段回落率及3周后左室射血分数进行比较.结果 用药后观察组患者的无复流发生率17.07％(7/41)低于对照组41.86％(18/43);观察组患者ST段回落率90.24％(37/41)高于对照组67.44％(29/43);观察组1周后左室射血分数(75.321±11.782)％高于对照组(62.371±9.383)％,差异具有统计学意义(P<0.05).结论 STEMI患者PCI术中发生无复流情况后运用尼可地尔相对常规硝酸甘油注射的效果更好,更有利于患者恢复.

目的 探究血尿酸(SUA)、同型半胱氨酸(Hcy)、脑钠肽(BNP)、肌钙蛋白Ⅰ(cTnⅠ)与急性冠脉综合征(ACS)患者病变程度的相关性.方法 选取2017年1月～2018年1月我院收治的ACS患者120例,按疾病类型不同分为不稳定型心绞痛(UAP)组、非ST段抬高心肌梗死(NSTEMI)组和ST段抬高心肌梗死(STEMI)组,各40例,另选择40名同期健康体检者作为对照组;检测并比较四组血清SUA、Hcy、BNP和cTnⅠ水平,分析不同血管病变支数患者SUA、Hcy、BNP和cTnⅠ水平.结果 STEMI组SUA、Hcy、BNP和cTnⅠ水平均高于UAP和NSTEMI组,NSTEMI组SUA、Hcy、BNP和cTnⅠ水平均高于UAP组,STEMI组、UAP组和NSTEMI组SUA、Hcy、BNP和cTnⅠ水平均高于对照组,差异有统计学意义(P<0.05);3支病变患者SUA、Hcy、BNP和cTnⅠ水平高于双支病变和单支病变,且双支病变患者的SUA、Hcy、BNP和cTnⅠ水平高于单支病变,差异有统计学意义(P<0.05).结论 机体SUA、Hcy、BNP和cTnⅠ水平与ACS病情密切相关,且同时检测四种指标水平有助于进一步了解患者病情,值得临床应用.

目的 研究前列地尔对行急诊PCI急性心肌梗死患者术后心功能的影响.方法 选取我院心内科2017年2月～2018年2月行急诊PCI术的的急性心梗患者120例,将所有患者随机采分为观察组和对照组,每组各60例.对照组给予常规水化治疗;观察组在常规组的基础上术前30分钟给予0.9％氯化钠溶液100 ml+前列地尔10μg静脉滴注,1次/d,连续7 d.观察两组患者术后6 h、12 h及24 h测定血清肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)水平,术后24 h及7 d测定N-末端脑钠肽前体(NT-proBNP)、左心室舒张末期内径(LVEDD)、左心室射血分数(LVEF),PCI术后即刻造影结果:校正的TIMI帧数(CTFC).结果 术后6 h、12 h以及24 h,观察组CK和CK-MB水平均低于对照组,差异有统计学意义(P<0.05).术后24 h,两组NT-proBNP、LVEDD、LVEF比较,差异均无统计学意义(P>0.05).术后7 d,观察组NT-proBNP、LVEDD均低于对照组,LVEF高于对照组,差异均有统计学意义(P<0.05).PCI术后即刻观察组患者CTFC为(26.81±4.12)帧,低于对照组的(35.92±4.91)帧,差异有统计学意义(P<0.05).结论 前列地尔可明显改善急性心梗患者PCI术后的心功能.

目的 探讨和分析肝动脉化疗栓塞术(TACE)治疗原发性肝癌预后的相关因素.方法 回顾性分析我院收治的原发性肝癌患者200例的临床资料为研究对象,对其观察数据进行随访,采用Logrank检验预后单因素分析,多因素分析采用Cox回归分析,筛选出与其生存的显著相关因素.结果 经TACE治疗后患者1年、3年和5年生存率分别为55.0%、31.5%和13.5%.单因素分析结果中肝功能分级、肿瘤分型、肿瘤数目、肿瘤分期、TA C E治疗次数、有无乙肝肝硬化、有无门脉癌栓、有无动静脉瘘是影响预后的独立因素(P<0.05或P<0.01),年龄、性别、AFP值等与肝癌术后生存无相关性(P>0.05).Cox多因素回归分析显示乙肝肝硬化病史、是否有门脉癌栓、肿瘤分期和TA C E治疗次数对患者的预后具有显著影响(P<0.01).结论 肝硬化病史、肿瘤分期、门脉癌栓对肝癌的预后具有显著影响,是独立危险因素,TACE治疗次数对影响预后具有保护效果.

Objective To study the protective effects of phosphodies terase 5 shRNA [PDE5shRNA] on doxorubicin (DOX)-induced cardiomyopathy in mice.Methods A single intraperitoneal injection of doxorubicin 15 mg · kg-1 to induce cardiomyopathy to establish model.Male 10-week-old C57BL/6J mice were randomly assigned to three groups:model group (n =16);experimental group(DOX + PDE5shRNA,n =16):a single intraperitoneal injection of same dose of doxorubicin and being simultaneously treated with a single myocardial injection of PDE5shRNA 1 x 1010 particles;Normal group (n =10):a single intraperitoneal injection of same volume of saline.After two weeks of administration,left ventricular end-systolic diameter (LVESD),left ventricular end-distolic diameter (LVEDD),left ventricular ejection fraction (LVEF) and percent fractional shortening (％ FS) were evaluated by echocardiography.Cardiac specimens were then subjected for HE (Hematoxyum Eosin),Masson staining,computing for each group of cardiac cell size and fibrosis area.Levels of cyclic guanosine monophosphate (cGMP) and protein kinase G (PKG) in the myocardium were assayed by ELISA.The expression of myosin heavy chain (MHC),Troponin I and Desmin were evaluated by Western blot.Results Two weeks later of administration,indicators in experimental group and model group were as follows:LVESD were(2.13 ±0.10),(2.75 ±0.12)mm;LVEDD were(2.98 ± 0.10),(3.42 ± 0.12) mm;LVEF were (58.74 ± 1.40)％,(48.53 ± 1.50)％;％ FS were (28.52 ± 2.10) ％,(19.59 ± 1.67) ％;the transverse diameter of cardiomyocytes were (14.68 ± 0.42),(13.75 ±0.38)μm;cardiac fibrosis were (2.28 ±0.20)％,(5.10 ±0.35)％;the levels of cGMP expression were (0.23 ± 0.02),(0.06 ± 0.01) pg · mg-1;the levels of PKG were (0.21 ± 0.02),(0.10 ± 0.01) pg · mg-1;the myocardial expression of MHC were 1.55 ± 0.16,1.15 ± 0.22;the expression of troponin I were 1.32 ± 0.08,0.88 ±0.08;the expression of Desmin were 1.327 ± 0.512,1.103 ± 0.038;all the above data were significantly different compared between two groups (all P ＜ 0.05).Conclusion The protective effects of PDE 5shRNA on doxorubicin-induced cardiomyopathy,mitigatting doxorubicin-induced impairment of cardiac function in mice,significantly attenuatting doxorubicin-induced atrophic degeneration of cardiomyocyte and myocardial fibrosis through possible activating cGMP/PKG signaling pathway.

Objective To study the impact of adenoviral short hairpin RNA (shRNA) targeting phosphodiesterase 5 (PDE5 shRNA) on myo-cardial cells apoptosis at early stage post -myocardial infarction ( MI ).Methods MI was induced in mice by left coronary artery ligation.Mice were randomly assigned to test group [ n=10 , adenoviral vectors inserted with shRNA sequence for the inhibition of phosphodiesterase 5 ( PDE5 ) were injected intramyocardially to infarcted and border area ] and control group ( n =10 , injection with adenoviral vectors without therapeutic PDE5 shRNA ).One week post -MI, apoptosis was evaluated by TdT-mediated dUTP nick -end labeling ( TUNEL ) , proteins were extracted from the left ventricular of heart , level of PDE5 expression was detected using Western Bloting , the level of guanosine 3′, 5′-cyclic phosphate ( cGMP) or protein kinase G ( PKG) activity in the left ven-tricular of heart was evaluated by enzyme linked immunosorbent assay (ELISA).Results One week post -MI, compared to control group, test group had reduced number of apoptotic cells in infarct and periinfarct areas, and significantly reduced cardiomyocytes apoptosis in the periinfarct regions ( P<0.05 ).Compared to control group , the level of PDE5 was&nbsp;significantly decreased and the levels of cGMP and PKG were significantly increased in the test group ( P <0.05 ).Conclusion It the PDE5 shRNA has protective effect on acute myocardial infarction and significantly inhibit the apopto -sis of myocardial cell which may be closely related to the increased expression of cGMP and PKG .

OBJECTIVE:To investigate the therapeutic effects of erythropoietin sustained-release gelatin hydrogel microspheres (EPO-GHM) on a murine model of hindlimb ischemia and related mechanisms. METHODS:Fifty two ten weeks old male C57BL/6J mice were assigned to 5 groups: sham-operated group (the right femoral artery suture was passed through the right femoral artery but not tied, n=8); saline group (right femoral artery ligation and intramuscular injection of saline at a dose of 4 ml/kg into the right hind limb, n=12); EPO group(right femoral artery ligation and intramuscular injection of EPO at a dose of 5 000 IU/kg into the right hind limb, n=12), empty GHM group (right femoral artery ligation and intramuscular injection of empty GHM at a dose of 4 ml/kg into the right hind limb, n=8); EPO-GHM group(right femoral artery ligation and intramuscular injection of EPO-GHM at a dose of 5 000 IU/kg into the right hind limb, n=12). The blood flow ratio of ischemic limb (right)/nonischemic limb (left) was measured using a laser Doppler perfusion imager. After 8 weeks, immunohistochemical analysis were used to evaluate the vessel density (vessel density of CD31 positive), arteriole density(vessel density of α-smooth muscle actin(α-SMA) positive) and muscle area(HHF35 positive area). The proliferating index of vessels was evaluated by double immunofluorescent labeling to evaluate effect of EPO-GHM on angiogenesis of ischemia limb. Western blot was used to evaluate the protein expression of EPO receptor, protein kinase B(AKT), p-AKT, endothelial nitric oxide synthase(eNOS), p-eNOS and matrix metalloproteinase 2(MMP-2). RESULTS:(1) Eight weeks later, the blood flow ratio of ischemic limb/nonischemic limb was significantly higher in the EPO-GHM group compared with other groups(0.810±0.080, 0.563±0.051, 0.570±0.056 and 0.561±0.052 respectively, all P<0.05). (2) CD31 antibody positive and α-SAM antibody positive densities were higher in the EPO-GHM group compared with other groups(P<0.01 or 0.05). (3)HHF35 positive area in saline group, EPO group, empty GHM group and EPO-GHM group were smaller than that of sham-operated group(all P<0.05). HHF35 positive area in saline group, EPO group, empty GHM group and EPO-GHM group were similar(all P>0.05). (4)The proliferating index of vessels was higher in the EPO-GHM group compared with other groups(P<0.01 or 0.05). (5) Compared with other groups, the protein levels of EPO receptor, AKT, p-AKT, p-eNOS and MMP-2 were significantly higher in EPO-GHM group(P<0.01 or 0.05) and level of eNOS was similar among five groups(P>0.05). CONCLUSION:RESULTS from present study suggest EPO-GHM could improve blood perfusion of ischemia limb in mice through increasing capillary and arteriolar densities and these beneficial effects are possibly mediated by EPOR up-regulation and AKT/p-eNOS/MMP-2 signaling pathway activation.

Objective: To study the impact of PDE5 shRNA on myocardial cells apoptosis at early stage post-MI. Methods:Myocardial infarction(MI) was induced in mice by left coronary artery ligation. Mice were randomly assigned to test group(n=10), in which adenovirus vector with PDE5 sh RNA particles were injected along the anterior and posterior LV wall, and model group(n=10), in which adenovirus vector without PDE5 shRNA particles were injected in the same manner. One week post-MI, apoptosis was evaluated using the TUNEL method, Proteins were extracted from the left ventricular of heart, level of PDE5 expression was detected using Western Bloting, and level of cGMP or PKG activity in the left ventricular of heart was evaluated using ELISA. Results: One week post-MI, compared to model group, test group had reduced number of apoptotic cells in infarct and periinfarct areas, and significantly reduced cardiomyocytes apoptosis in the peri-infarct regions(P0.05). Compared to model group, the level of PDE5 significantly decreased and the levels of c GMP and PKG significantly increased in the test group(P0.05). Conclusions: The present study suggests the protective effects of PDE5 sh RNA on acute myocardial infarction, significantly inhibiting myocardial cell apoptosis through possible activating cGMP/PKG signaling pathway.

Introduction Treatment with short hairpin RNA (shRNA) interference therapy targeting phosphodiesterase 5a after myocardial infarction (MI) has been shown to mitigate post-MI heart failure. We investigated the mechanisms that underpin the beneficial effects of PDE5a inhibition through shRNA on post-MI heart failure. Methods An adenoviral vector with an shRNA sequence inserted was adopted for the inhibition of phosphodiesterase 5a (Ad-shPDE5a) in vivo and in vitro. Myocardial infarction (MI) was induced in male C57BL/6J mice by left coronary artery ligation, and immediately after that, the Ad-shPDE5a was injected intramyocardially around the MI region and border areas. Results Four weeks post-MI, the Ad-shPDE5a-treated mice showed significant mitigation of the left ventricular (LV) dilatation and dysfunction compared to control mice. Infarction size and fibrosis were also significantly reduced in Ad-shPDE5a-treated mice. Additionally, Ad-shPDE5a treatment decreased the MI-induced inflammatory cytokines interleukin (IL)-1β, IL-6, tumor necrosis factor-α, and transforming growth factor-β1, which was confirmed in vitro in Ad-shPDE5a transfected myofibroblasts cultured under oxygen glucose deprivation. Finally, Ad-shPDE5a treatment was found to activate the myocardial Akt signaling pathway in both in vivo and in vitro experiments. Conclusion These findings indicate that PDE5a inhibition by Ad-shPDE5a via the Akt signal pathway could be of significant value in the design of future therapeutics for post-MI heart failure.

OBJECTIVE:To observe the impact of PDE5shRNA on cardiac remodeling and heart function following myocardial infarction in mice.METHODS:Myocardial infarction (MI) was induced in mice by left coronary artery ligation. Mice were randomly assigned to sham group (n = 6), PDE5shRNA group (n = 12), common adenovirus group (n = 15) and DMEM group (n = 8). Four weeks post-MI, the survival rate was evaluated. Cardiac function was examined by echocardiography. HE staining and Masson staining were used to evaluate the myocardial infarction size and fibrosis. The number of blood vessels was evaluated by immunohistochemistry, PDE5 protein expression in the left ventricular was detected using Western blot, level of cGMP or PKG activity in the left ventricle was evaluated with ELISA.RESULTS:Four weeks post-MI, all mice survived in the sham group, 3(37%) mice died in the DMEM group, 1 (8%) died in the PDE5shRNA group and 5 died in the common adenovirus group (33%). Infarct size was significantly reduced in PDE5shRNA group compared with the common adenovirus group and DMEM group [(25.4 ± 2.9)% vs. (42.0 ± 3.2)% and (43.4 ± 2.6) %, P < 0.05]. Cardiac function was significantly improved in PDE5shRNA group compared to common adenovirus group and DMEM group[LVFS: (21.1 ± 3.7)% vs. (14.2 ± 2.9)% and (14.22 ± 2.91)%, all P < 0.05; LVEF: (48.2 ± 7.1)% vs. (34.6 ± 6.2)% and (38.1 ± 2.8)%, all P < 0.05; LVESD: (3.87 ± 0.45) mm vs.(4.91 ± 0.62) mm and (4.63 ± 0.37) mm, all P < 0.05]. The blood vessel density was also higher in PDE5shRNA group compared with common adenovirus group (infarct area:14.3 ± 2.0 vs. 6.6 ± 1.2, P < 0.05; periinfarct area: 23.6 ± 2.1 vs. 13.7 ± 2.4, P < 0.05). Compared with common adenovirus group, level of PDE5 was significantly downregulated and level of cGMP or PKG was significantly upregulated in PDE5shRNA group (all P < 0.05).CONCLUSIONS:Present study suggests PDE5shRNA improves cardiac function and attenuates cardiac remodeling through reducing infarction size and cardiac fibrosis and these beneficial effects are possibly mediated by activating cGMP/PKG signaling pathway.

目的 观察磷酸二酯酶5(PDE5) shRNA对缺氧缺糖条件下乳鼠心肌细胞的保护作用.方法 培养新生C57BL/6J小鼠心肌细胞,通过缺氧缺糖建立小鼠心肌细胞缺氧缺糖损伤模型.随机分2组:实验组(shPDE5)通过构建PDE5 shRNA,将PDE5 shRNA转染心肌细胞;对照组(Null):将普通腺病毒载体转染心肌细胞.用原位末端标记分析(TUNEL)检测2组细胞凋亡情况;免疫蛋白印迹检测PDE5蛋白的表达;用酶联免疫吸附法测定各组心肌细胞cGMP和PKG的水平.结果 与对照组比较,实验组PDE5表达明显下降,cGMP和PKG的活性明显上调(P<0.05);实验组细胞凋亡数明显减少(P<0.05).结论 PDE5 shRNA可明显拮抗缺氧缺糖诱导的心肌细胞凋亡,可能与PDE5 shRNA持续抑制心肌细胞PDE5基因表达、显著上调cGMP和PKG活性有关.

冠状动脉分叉病变占总的经皮冠状动脉介入治疗（PCI）的15％～20％，探索新的有效的冠状动脉分叉病变治疗技术仍然是一条很漫长的路[1-2]。分叉病变的介入治疗是介入治疗领域的一个难点，其成功率低，主要不良心脏事件［包括围术期心肌梗死（MI）、靶病变血运重建（TLR）、支架内血栓］发生率高[3-5]。因此，分叉病变介入治疗技术的关键是同时保护主支和边支，降低围术期并发症的发生率及减少TLR。

BACKGROUND:It is not clear yet how tadalafil affects nonischemic cardiomyopathy, although its beneficial effects on acute myocardial infarction are well-known. We investigated tadalafil's beneficial effects on nonischemic cardiomyopathy and the specific mechanisms of its effects.METHODS:Cardiomyopathy was induced in mice by a single intraperitoneal injection of doxorubicin (15 mg/kg). In some cases, tadalafil (4 mg/kg/day, p.o., 14 days) was started simultaneously. After two weeks, cardiac function was evaluated by echocardiography and cardiac catheterization, then all of the mice were killed and cardiac specimens were subjected for hemotoxylin and eosin staining, Masson's trichrome staining, terminal deoxynucleotidyltransferase dUTP nick-end labeling assay, enzyme-linked immunosorbent assay, and Western blot.RESULTS:Two weeks later, left ventricular dilatation and dysfunction were apparent in mice given doxorubicin but were significantly attenuated by tadalafil treatment. Tadalafil also protected hearts against doxorubicin-induced cardiomyocyte atrophy/degeneration and myocardial fibrosis. No doxorubicin-induced apoptotic effects were seen between groups. Cardiac cGMP level was lower in the doxorubicin-treated group, however it was significantly increased with tadalafil treatment. Compared to the control group, the myocardial expression of 3 sarcomeric proteins, myosin heavy chain, troponin I, and desmin were significantly decreased in the doxorubicin-treated group, which were restored by the tadalafil treatment.CONCLUSIONS:The present study indicates a protective effect of tadalafil mainly through cGMP signaling pathway against doxorubicin-induced nonischemic cardiomyopathy.

目的 探讨抑制磷酸二酯酶5的短发夹RNA重组腺病毒载体PDE5 shRNA对缺氧缺糖条件下乳鼠心肌细胞的保护作用.方法 培养新生C57BL/6J小鼠心肌细胞,通过缺氧缺糖,建立小鼠心肌细胞缺氧缺糖损伤模型,分实验组(shPDE5)和对照组(Null),实验组:通过构建抑制磷酸二酯酶5的短发夹RNA重组腺病毒载体(PDE5 shRNA),将PDE5 shRNA转染心肌细胞,对照组(Null):将没有插入PDE5 shRNA的普通腺病毒载体转染心肌细胞.利用TUNEL法检测各组细胞凋亡情况.结果 与对照组比较,细胞在缺氧缺糖条件下,实验组细胞凋亡明显减少(P<0.05).结论 PDE5 shRNA可明显拮抗缺氧缺糖诱导的心肌细胞凋亡.

Objective To study the protective effects and its mechanisms of tadalafil on doxorubicin-induced cardiomyopathy in mice.Methods Mice were randomly assigned to three groups: model and test group(both n=16): a single intraperitoneal injection of doxorubicin 15 mg·kg-1 to induce cardiomyopathy;normal group(n=10): a single intraperitoneal injection of same volume of saline.On that basis,test group was treated with tadalafil 4 mg·kg-1,once a day,for 14 days,orally viagauge.After two weeks,echocardiograms were then recorded,histological analysis and the cardiac cGMP level were detected.Results Two weeks later,indicators of test group and model group were as follows: left ventricular end-diastolic dimension were(3.07±0.10),(3.50±0.20)mm;left ventricular end-systolic dimension were(2.17±0.10),(2.82±0.17)mm;left ventricular ejection fraction were(57.85±2.03)%,(46.07±2.96)%;left ventricular percent fractional shortening were(29.20±3.05)%,(19.41±2.59)%;the transverse diameter of cardiomyocytes were(14.24±0.39),(13.78±0.36)μm;cardiac fibrosis were(2.35±0.21)%,(5.16±0.36)%;the levels of cGMP were(0.15±0.01),(0.05±0.01)pg·mg-1.There were significiant compared with model group(P<0.05).The myocardial expression of myosin heavy chain,troponin I and desmin were significantly increased in the test group.No significant difference apoptotic effects were seen between each group.Conclusion The protective effects of tadalafil against doxorubicin-induced cardiomyopathy,mitigatting doxorubicin-induced impairment of cardiac function in mice,significantly attenuatting doxorubicin-induced atrophic degeneration of cardiomyocyte and myocardial fibrosis through possible activating cGMP signaling pathway.

BACKGROUND:Adverse cardiac event rates are higher for percutaneous coronary intervention (PCI) of bifurcation lesions. Currently, provisional stenting or a simple stenting strategy is the standard treatment for bifurcated lesions, but its performance remains limited because of a risk of side-branch (SB) closure and a higher rate of target lesion revascularization (TLR). We report a new provisional side-branch stenting strategy to treat coronary bifurcation lesions using a "balloon-stent kissing" technique (BSKT).METHODS:From January 2011 to December 2012, a total of 60 patients with 60 bifurcation lesions underwent PCI using BSKT. Baseline and postprocedural quantitative coronary angiography (QCA) analyses were performed. Procedural and immediate clinical outcomes were reviewed.RESULTS:The majority of patients presented with acute coronary syndrome (98%) and had true bifurcation lesions (98%). TIMI-3 flow was established in 100% of the main branch and SB lesions. QCA revealed preservation of the bifurcation angle after PCI (pre PCI, 57.0 ± 16.3; post PCI, 60.5 ± 16.1; P=.24). Five patients (8%) had lesions that required rewiring and 2 patients (3%) required provisional stenting of the SB. No SB loss occurred during PCI. No patient had a periprocedural myocardial infarction, nor was the balloon or wire entrapped during any PCI.CONCLUSIONS:As a new coronary bifurcation provisional stenting method, the BSKT is associated with a high procedural success rate, improved SB patency, and a low rate of immediate cardiac events.