ADAMTS-4在心力衰竭患者心肌中的表达及意义

Expression and significance of ADAMTS-4 in myocardium in patients with heart failure

目的探讨含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶-4(ADAMTS-4)在心力衰竭患者心肌中的表达及作用。方法收集2012年7月~2017年3月武汉大学人民医院终末期心力衰竭患者心脏移植后的病变心脏及配型不成功的正常供体心脏标本,心力衰竭患者17例,纳入心力衰竭组,男性12例,女性5例,年龄(46.5±9.7)岁;配型不成功的正常供体心脏8例,纳入对照组,均为男性,年龄(37.6±6.9)岁。心脏离体后立即分离样本,心肌组织采用HE和天狼星红染色,采用Real-time PCR及Western-blot技术检测ADAMTS-4在心肌组织中的表达水平。结果光镜下,对照组心肌细胞形态完整,胞浆均匀致密,细胞排列整齐,可见清晰的细胞核;心力衰竭组患者心肌细胞肿大,形态不规整,肌丝断裂,排列紊乱,可见畸形、增大的细胞核。天狼星红染色显示,心力衰竭组患者心肌组织胶原纤维含量较对照组增多,胶原纤维增粗、排列紊乱,胶原容积分数较对照组升高,(10.18±0.63)%vs.(2.56±0.25)%,差异具有统计学意义(P<0.01)。与对照组相比,心力衰竭组患者心肌组织ADAMTS-4m RNA相对表达水平明显增高,(1.00±0.03)vs.(1.50±0.03),差异具有统计学意义(P<0.01)。与对照组相比,心力衰竭组患者心肌组织ADAMTS-4蛋白表达水平明显升高,(1.00±0.06)vs.(2.01±0.09),差异具有统计学意义(P<0.01)。结论含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶-4可能通过参与心肌组织重构影响心力衰竭的发生。

Objective To investigate the expression and effect of a disintegrin and metalloprotease with thrombospondin motifs-4 （ADAMTS-4） in myocardium in patients with heart failure （HF）. Methods The heart specimens （disease heart and normal heart failed in matching） were collected from HF patients （n=17, male 12, female 5 and average age=46.5±9.7, HF group）, and male donators （n=8 and average age=37.6±6.9, control group） from People’s Hospital of Wuhan University from July 2012 to Mar. 2017. The samples of myocardial tissue were given HE staining and picrosirius staining, and expression of ADAMTS-4 was detected by using real-time fluorescence quantitative polymerase chain reaction （RT-PCR） and Western-blot method. Results In the control group, integral morphology, uniform and compact plasma and alignment of myocardial cells with clear cell nucleus were observed under light microscope. In HF group, edema, irregular morphology, broken myofilament, disordered arrangement, and malformed and enlarged cell nucleus were observed in myocardial cells. The results of picrosirius staining showed that content of collagen fibers increased with thickened and irregular collagen fibers, and collagen volume fraction （CVF） increased [（10.18±0.63）% vs. （2.56±0.25）%] in HF group compared with control group （P〈0.01）. The relative expression of ADAMTS-4 mRNA increased significantly in HF group compared with control group [（1.00±0.03） vs. （1.50±0.03）, P〈0.01]. The expression of ADAMTS-4 protein increased significantly in HF group compared with control group [（1.00±0.06） vs. （2.01±0.09）, P〈0.01]. Conclusion ADAMTS-4 may influence HF occurrence through participating myocardial remodeling.