早期体外心脏震波通过抑制TGF-β1-Smad2/3-MMP-9信号通路改善心肌纤维化
Early extracorporeal cardiac shock wave inhibits myocardial fibrosis by inhibiting TGF-β1-Smad2/3-MMP-9 signaling pathway
李纯 1王露 1叶雨佳 1杨妍 1宋文娟 1马雪娟 1刘利萍 1谷颍 1赵月 1孙钺 1王钰1
作者信息
- 1. 650032 昆明医科大学第一附属医院心内科
- 折叠
摘要
目的 探究急性心肌梗死(AMI)大鼠早期经体外心脏震波治疗(CSWT)后心功能和心肌纤维化改善与转化生长因子β1-细胞信号转导分子2/3-基质金属蛋白酶9(TGF-β1-Smad2/3-MMP-9)信号通路的相关性.方法 采用随机数字表法将30只SD大鼠分为sham、AMI、AMI+CSWT、AMI+SB431542和AMI+SB431542+CSWT组,每组各6只.sham组仅开胸,其他4组构建AMI模型.AMI+SB431542 组和 AMI+SB431542+CSWT 组在术后 4 d 行 SB431542 处理,持续 3 d.AMI+CSWT 组和AMI+SB431542+CSWT组均行9次CSWT[于术后7 d开始行CSWT,两次之间间隔1 d,能量:0.09 mJ/mm2,一个缺血区域共1 800震,频率:3 Hz(3震/s)].各组均在术后4 d和62 d行超声心动图检测左心室射血分数(LVEF)、左心室短轴缩短率(LVFS)、左心室舒张末期容积(LVEDV)和左心室收缩末期容积(LVESV).术后62 d行心脏组织取材,HE染色、Masson染色评估心肌组织病理形态学改变及心肌纤维化程度;于心肌梗死边缘区检测TGF-β1-Smad2/3-MMP-9信号通路相关蛋白表达水平及COL-Ⅰ、COL-Ⅲ蛋白表达水平.结果 (1)术后4 d,与sham组相比,AMI、AMI+CSWT、AMI+SB431542 和 AMI+SB431542+CSWT 组的 LVEF、LVFS 明显降低,LVESV 明显扩大(均为 P＜0.05)a(2)术后62 d,与sham组相比,AMI组的LVEF、LVFS明显降低,LVEDV、LVESV明显扩大(均为P＜0.05);与AMI组相比,AMI+CSWT组的LVEF、LVFS明显改善,LVEDV、LVESV明显回缩(均为P＜0.05);与 AMI+CSWT 组相比,AMI+SB431542+CSWT 组的 LVEF、LVFS、LVEDV 和 LVESV 无统计学差异(均为P＞0.05).(3)HE染色显示,AMI组较sham组红色区域排列紊乱、蓝色区域增多;AMI+CSWT组较AMI组红色区域排列整齐、蓝色区域减少,但较sham组蓝染区域增多.(4)Masson染色显示,与sham组相比,AMI组胶原容积分数(CVF)明显升高(P＜0.001);与AMI组相比,AMI+CSWT和 AMI+SB431542 组的 CVF 明显降低(均为 P＜0.05);与 AMI+CSWT 组相比,AMI+SB431542+CSWT组CVF较高,但无统计学差异(P=0.65).(5)Western blot结果显示,与sham组相比,AMI组的TGF-β1、Smad3、MMP-9、COL-Ⅰ和COL-Ⅲ表达水平明显升高(均为P＜0.05);Smad2水平有升高趋势,但无统计学差异(P=0.40);与 sham 组和 AMI 组相比,AMI+CSWT 组的 TGF-β1、Smad2、Smad3、MMP-9、COL-Ⅰ 和 COL-Ⅲ表达水平明显降低(均为 P＜0.05);与 AMI+CSWT 组相比,AMI+SB431542+CSWT组的TGF-β1、Smad2、Smad3和MMP-9表达水平明显降低,COL-Ⅰ和COL-Ⅲ表达水平明显增加(均为P＜0.05).结论 早期CSWT可改善心功能、抑制心肌纤维化,且与抑制TGF-β1-Smad2/3-MMP-9信号通路相关.
Abstract
Objective To investigate the correlation between the improvement of cardiac function and myocardial fibrosis and the transforming growth factor β1-cellular signaling molecule 2/3-matrix metalloproteinase 9(TGF-β1-Smad2/3-MMP-9)signaling pathway in acute myocardial infarction(AMI)rats after early extracorporeal cardiac shock wave therapy(CSWT).Methods Thirty SD rats were divided into sham group,AMI group,AMI+CSWT group,AMI+SB431542 group or AMI+SB431542+CSWT group by using randomized numerical table method with 6 rats in each group.Rats in sham group only received open thorax surgery,and the other 4 groups were constructed with AMI model.Rats in AMI+SB431542 group and the AMI+SB431542+CSWT group were treated with SB431542 for 3 days.The AMI+CSWT group and the AMI+SB431542+CSWT group were treated with 9 times of CSWT(CSWT was performed on the 7th postoperative day with the energy of 0.09 Mj/mm2,the number of times of CSWT:1 800 times,and the frequency of CSWT:3 Hz(3 times/s)).Echocardiography was performed at 4 and 62 d after surgery to detect left ventricular ejection fraction(LVEF),left ventricular short-axis shortening(LVFS),left ventricular end-diastolic volume(LVEDV),and left ventricular end-systolic volume(LVESV)in each group.At 62 d after surgery,cardiac tissue was sampled,and HE staining and Masson staining were used to evaluate the pathological changes in myocardial tissue and the degree of myocardial fibrosis;TGF-β1-Smad2/3-MMP-9 signaling pathway-related protein expression levels and COL-Ⅰ and COL-Ⅲ protein expression levels were detected in the infarcted border area.Results(1)At 4 d postoperatively,compared with the sham group,LVEF and LVFS were significantly lower and LVESV was significantly enlarged in the AMI group,AMI+CSWT group,AMI+SB431542 group,and AMI+SB431542+CSWT group(all P＜0.05).(2)At 62 d postoperatively,LVEF and LVFS were significantly lower and LVEDV and LVESV were significantly enlarged in the AMI group compared with the Sham group(all P＜0.05);LVEF and LVFS were significantly improved and LVEDV and LVESV were significantly retracted in the AMI+CSWT group compared with the AMI group(all P＜0.05);there were no statistically significant differences in LVEF,LVFS,LVEDV,and LVESV in the AMI+SB431542+CSWT group compared with the AMI+SB431542+CSWT group(all P＞0.05).(3)HE staining showed that the red areas were disorganized and the blue areas increased in the AMI group compared with the sham group;the red areas were neatly arranged and the blue areas decreased in the AMI+CSWT group compared with the AMI group,but the blue-stained areas increased in the sham group.(4)Masson staining showed that the collagen volume fraction(CVF)was significantly higher in the AMI group compared with the sham group(P＜0.001);the CVF was significantly lower in the AMI+CSWT group and AMI+SB431542 group compared with the AMI group(both P＜0.05);and the CVF was higher in the AMI+SB431542+CSWT group compared with the AMI+CSWT group,but did not show statistical difference(P=0.65).(5)Western blot showed that compared with the sham group,the expression levels of TGF-β1,Smad3,MMP-9,COL-Ⅰ and COL-Ⅲ were significantly higher in the AMI group(all P＜0.05);there was a mild elevation of Smad2 without significant difference(P=0.40);compared with the sham group and the AMI group,the expression levels of TGF-β1,Smad2,Smad3,MMP-9,COL-Ⅰ and COL-Ⅲ were significantly lower in the AMI+CSWT group(all P＜0.05);compared with AMI+CSWT group,the expression levels of TGF-β1,Smad2,Smad3 and MMP-9 were significantly lower and COL-Ⅰ and COL-Ⅲ were significantly increased in the AMI+SB431542+CSWT group(all P＜0.05).Conclusions Early CSWT can improve cardiac function and inhibit myocardial fibrosis,and may be associated with the TGF-β1-Smad2/3-MMP-9 signaling pathway.
关键词
体外心脏震波治疗/大鼠/急性心肌梗死/心脏功能/心肌纤维化Key words
Extracorporeal cardiac shock wave therapy/Rats/Acute myocardial infarction/Cardiac function/Myocardial fibrosis引用本文复制引用
出版年
2024
NETL
NSTL
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