摘要
目的探讨色素上皮衍生因子(PEDF)基因修饰骨髓干细胞(MSC)对糖尿病心肌梗死大鼠心肌损伤的作用及其对Wnt/连环蛋白(β-catenin)信号通路相关蛋白表达的影响。方法SPF级SD大鼠40只,随机均为分4组:糖尿病组(DM组)、糖尿病心肌梗死模型组(DMI组)、给予沉默PEDF的MSC糖尿病心肌梗死组(PM组)以及给予沉默PEDF的MSC和Wnt/β-catenin通路抑制剂的糖尿病心肌梗死组(WPM组)。采用高脂喂养+腹腔注射链脲佐菌素(STZ)联合结扎大鼠冠状动脉建立糖尿病心肌梗死模型。PM组于模型成功后移植PEDF-/MSC(50μL)。WPM组于模型成功后移植PEDF-/MSC(50μL),30 min后静脉注射HY-15597(8 mg/kg)。观察大鼠心功能情况;HE染色观察大鼠心脏的病理变化;免疫荧光检测各组大鼠心肌组织中血管生成因子VEGF、IGF-1、PDGF B的表达;Western blotting检测各组大鼠心肌组织中Wnt/β-catenin信号通路相关蛋白的表达。结果术后24 h,PM组CK、CK-MB、BNP水平明显低于DMI组(P<0.05),术后4周CK、CK-MB、BNP水平检测结果显示,与24 h结果比较DMI组和PM组各项指标均下降(P<0.05)。与DMI组比较,超声提示PM组大鼠左室射血分数(LVEF)明显升高(P<0.05);病理学检查PM组可见心肌细胞改善,损伤程度有所好转,心肌细胞形态尚完整,肌纤维排列有序,心肌组织充血、肿胀减轻。免疫荧光提示沉默PEDF的MSC可以促进血管生成因子表达,同时显著下调wnt3a、β-catenin、survivin的表达(P<0.05)。结论沉默PEDF的MSC可通过促进血管生成改善糖尿病大鼠心肌梗死的心功能,对心脏具有保护作用。
Objective To investigate the effect of pigment epithelium-derived factor(PEDF)gene modified marrow stem cells(MSC)on myocardial injury in diabetic rats with myocardial infarction and on the expression of Wnt/beta-catenin signaling pathway-related proteins.Methods Forty SPF grade SD rats were randomly divided into the following four groups:diabetic group(DM group),diabetic myocardial infarction model group(DMI group),MSC diabetic myocardial infarction group(PM group;in which PEDF was silenced),and diabetic myocardial infarction group(WPM group)administered with MSC in which PEDF was silenced and Wnt/beta-catenin pathway inhibitors.Using high fat feeding urea+intraperitoneal injection of chain with streptozotocin(STZ)rats coronary artery ligation of diabetic myocardial infarction model was established.The PM group was successfully transplanted with PEDF-/MSC(50μL).The cardiac function of rats and examination related was observed.Results After 24 h and 4 weeks post-surgery,the levels of CK,CK-MB,and BNP in the PM group were significantly lower than those in the DMI group(P<0.05).Compared with the DMI group,left ventricular ejection fraction(LVEF)in the PM group was significantly high(P<0.05).Immunofluorescence indicated that MSC in which PEDF was silenced,could promote the expression of angiogenic factors(P<0.05),and could significantly downregulate the expression of Wnt3a,β-catenin,and survivin(P<0.05).Conclusion MSC with silenced PEDF can improve heart function in diabetic rats with myocardial infarction by promoting angiogenesis.
作者
刘宇
石云
张旭
高妍
王欢
王斌
LIU Yu;SHI Yun;ZHANG Xu;GAO Yan;WANG Huan;WANG Bin(Department of Cardiovascular Surgery,General Hospital of Northern Theater Command,Shenyang 110016,China)
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2019年第12期1091-1095,1099,共6页
Journal of China Medical University
基金
辽宁省自然科学基金（201601391）
全军医学科技青年培育项目（16QNP056）
关键词
色素上皮衍生因子
骨髓干细胞
糖尿病心肌梗死
大鼠
心肌损伤
信号通路
pigment epithelial derived factor
bone marrow stem cell
diabetic myocardial infarction
rat
myocardial injury
signaling pathway
