改良法分离培养SD新生乳鼠原代心肌细胞

目的:建立一种稳定、快速的SD新生乳鼠原代心肌细胞分离培养改良方法。方法:取SD新生乳鼠心室,0.12%Ⅱ型胶原酶消化,Percoll密度梯度离心结合5-溴脱氧尿嘧啶(5-BrdU)化学抑制法纯化心肌细胞,体外培养于含5%马血清的改良DMEM/F12中,次日更换为普通含10%胎牛血清的高糖DMEM继续培养,并比较此改良方法与传统差速贴壁法的差异。结果:改良法获得的心肌细胞生长良好,接种24 h后几乎全部贴壁生长,细胞呈三角形、梭形或不规则形,个别细胞出现自主搏动,频率为10~30 beats/min不等。48 h后心肌细胞变长伸出伪足,部分细胞呈现同步搏动,频率接近50~80 beats/min。72 h后心肌细胞成菊花样交织成网,自发搏动趋于同步,频率加快至80~100 beats/min;96 h后细胞聚集成簇,呈岛屿样,同步搏动频率在100~120 beats/min左右,一周内细胞状态良好。改良法纯化原代心肌细胞的得率((1.17±0.15)×10^(6) vs(1.21±0.22)×10^(6),P>0.05)和存活率与传统差速贴壁法相当(93.3%±1.4%vs 92.2%±0.7%,P>0.05),但是改良方法获得的原代心肌细胞纯度更高(94.7%±2.1%vs 89.5%±1.3%,P<0.05),且用时较短((3.1±0.4)h vs(4.3±0.3)h,P<0.01)。结论:改良法获得心肌细胞耗时短、纯度高、结构功能保存完整,且实验重复和稳定性好,是一种理想且简单易行的的原代心肌细胞分离培养方法。

择期冠状动脉介入治疗后CTnI与GLS、QTd的关系

目的探讨老年患者行择期冠状动脉介入治疗(PCI)后肌钙蛋白I(CTnI)与QT离散度(QTd)、左室长轴应变(GLS)的关系。方法选取2018年3月至2019年7月入住宁波市某公立三甲综合医院心血管内科且年龄≥60岁的冠心病患者为研究对象,检测PCI前后患者肌钙蛋白I(术前1 d及术后24 h)、QTd及GLS(均为术前及术后4个周)。根据PCI术后CTnI数值,分为两组:CTnI(+)组(n=26)及CTnI(-)组(n=104)。同时测定PCI前、后两组的QTd、GLS,分析两组PCI前后QTd、GLS的变化。结果CTnI(+)组手术前后QTd分别为(52.3±16.2)ms和(46.1±12.5)ms,组内比较,差异有统计学意义(P<0.01);CTnI(-)组手术前后的QTd分别为(53.5±15.9)ms和(41.3±12.9)ms,组内比较,差异有统计学意义(P<0.001),组间比较,差异有统计学意义(P<0.05)。CTnI(+)组PCI术前、后GLS分别为(-18.42±2.21)%和(-20.37±2.67)%,CTnI(-)组PCI术前、后GLS分别为(-19.38±2.88)%和(-22.21±2.63)%,PCI后组间比较,差异有统计学意义(P<0.05)。结论老年患者择期冠状动脉介入治疗(PCI)后出现CTnI阳性者,其QTd缩短幅度变小,GLS改善幅度较小,表明其存在亚临床心功能受损,提示可能增加患者择期冠状动脉介入治疗术后主要心脏不良事件发生。

The effect of spironolactone on the levels of GATA4 protein and hypoxia inducible factor-1α in patients receiving coronary stent implantation

In the present study,we aimed to explore the protective effect of spironolactone on cardiac function in patients undergoing coronary stent implantation by determining the serum levels of GATA4 and hypoxia-inducible factor-1α(HIF-1α)proteins before and after the coronary stent implantation.A total of 134 patients undergoing coronary stent implantation in our hospital from March 2019 to March 2020 were retrospectively selected using the propensity score matching(PSM)method.Of the 134 patients,67 patients taking spironolactone were used as a test group,and the other 67 patients without taking spironolactone were used as a control group.In all patients,the levels of serum GATA4,HIF-1α,and troponin I proteins were determined before as well as 24 h and 6 months after the coronary stent implantation.Left ventricular ejection fraction(LVEF)and left ventricular systolic global longitudinal strain(GLS)were determined before and 6 months after the coronary stent implantation.There were no significant differences in the HIF-1αlevel between the two groups before and 6 months after the operation,while the HIF-1αlevel was significantly lower in the test group compared with the control group at 24 h after the operation(P<0.01).There were no significant differences in the GATA4 protein level between the two groups before and 24 h after the operation,while the GATA4 protein level was significantly lower in the test group compared with the control group at 6 months after the operation(P<0.01).There was no significant difference in LVEF between the two groups before and 6 months after the operation.GLS was significantly improved at 6 months after the operation compared with that before the operation in both groups,while GLS was significantly better in the test group compared with the control group at 6 months after the operation(P<0.01).Collectively,spironolactone could protect cardiac function probably by improving myocardial hypoxia and inhibiting myocardial remodeling.