间充质干细胞过表达IL-10对心肌梗死模型大鼠心功能的影响

Effect of mesenchymal stem cells overexpressing il10 transplanted in rats with myocardial infarction

目的探究间充质干细胞(MSC)转运白介素-10(IL-10)后对其心功能的影响及影响机制。方法用SD大鼠制造心肌梗死(MI)模型,构建真核质粒载体后随机分为C组(注射MSC与PBS混合悬液)、P组(注射pc DNA3-IL-10的MSC悬液);K组(注射pc DNA3的MSC悬液)。通过心脏超声检查、血流动力学检查和组织学染色方法分别检测大鼠左心室射血分数、短轴缩短率、左心室收缩末压力、左心室舒张末压力、左心室压最大升降速度及梗死面积;免疫荧光检测各组大鼠心肌细胞Caspase-3蛋白含量,同时,western blot检测炎性因子TNF-α、IL-1β含量。结果各组在不同时间点下,三组射血分数、左室短轴缩短率随时间变化并无明显差异(P>0.05);C、K两组各点数值相近,无统计学差异,P组各时间点下射血分数、左室短轴缩短率远大于前两组(P<0.05);组间对比,P组射血分数、左室短轴缩短率均显著高于K组、C组(P<0.05);各组射血分数(F_(交互)=2.564,P_(交互)=0.015)、左室短轴缩短率(F_(交互)=2.233,P_(交互)=0.022)均存在交互作用。术后4周,P组LVSP、+dp/dtmax、-dp/dtmax显著高于C组、K组,而LVEDP则明显低于前两组(P<0.01),其梗死区域最低;免疫荧光及western blot结果显示P组Caspase-3、TNF-α、IL-1β蛋白含量较C、K两组明显降低,差异有统计学意义(P<0.05)。结论MSC转导IL-10可促进MI发生后心功能的恢复,这可能与IL-10过表达抑制Caspase-3凋亡基因及TNF-α、IL-1β炎性因子的表达相关。

Objective To investigate the modulation effects of mesenchymal stem ceils （MSCs） overex- pressing IL-10 transplanted in a rat model of myocardial infarction and its possible mechanism. Methods The MI rats were established by left anterior descending coronary artery ligation and the rats were then randomly divided into three groups： group C （MSC ＋ PBS）, group P （pcDNA3-IL-10＋MSC）, group K （peDNA3 ＋ MSC）. Eehocardiography and hemodynamic examinations were used to evaluate the cardiac function. Myocardial infarction size were evaluate were evaluate by Immumohistochemical stainingmyocardial. At the same time, Immunofluorescence and western blot was applied to show the expression of Caspase-3, TNF-α and 1L- 1β, respectively. Results The left ventricular ejection fraction and fractional shortening in three groups showed no significant difference （P 〉 0.05 ） at different time; There were no statistically significant differences between the groups K and group C and the left ventricular ejection fraction, fractional shortening in group P were highest （P 〈 0.05） ; The left ventricular ejection fraction （Finteractive = 2.564, Pinteractive = 0.015） and fractional shortening （Finteractive = 2.233, Pinteractive= 0.022） have interactive effect in three groups. After 4 weeks, LVSP, ＋dp/dtmax and -dp/dtmax in group P were significantly higher than that of C group and K group, while the LVEDP was lower （P 〈 0.01） ; immunofluorescence and Western blot showed that Caspase-3, TNF-α and IL-1β in group P were significantly lower than that of C group and K group, and the difference was statistically significant （P 〈 0.05）. Conclusion MSC overexpressing IL-10 can promote the recovery of cardiac function after MI, which may be related to inhibition of Caspase-3 apoptosis gene and TNF-α and IL- 1β inflammatory factors.