浙江中西医结合杂志

血府逐瘀汤通过抑制JAK/STAT信号通路调节线粒体自噬改善心梗后心室重构的机制研究

Mechanistic Study of Xuefu Zhuyu Decoction in Improving Post-Myocardial Infarction Ventricular Remodeling via Regulation of Mitophagy through Inhibition of the JAK/STAT Signaling Pathway

投稿时间：2025-08-27 修订日期：2025-12-17

DOI：

中文关键词: 血府逐瘀汤心室重构 JAK/STAT信号通路线粒体自噬

英文关键词:Xuefu Zhuyu Decoction ventricular remodeling JAK/STAT signaling pathway mitochondrial autophagy

基金项目:浙江省中医药科技计划项目(2024ZL636)

作者	单位	邮编
周佳^*	浙江中医药大学附属杭州市中医院	310007
张勇	浙江中医药大学附属杭州市中医院
陈远园	浙江中医药大学附属杭州市中医院
葛美芳	浙江中医药大学附属杭州市中医院

摘要点击次数: 0

全文下载次数: 0

中文摘要:

目的：体内外实验探究血府逐瘀汤（XFZYD）通过调节线粒体自噬改善心梗（MI）后心室重构的作用机制。方法：将SD大鼠分为三组：假手术组、模型组和XFZYD组。通过结扎冠脉左前降支构建MI大鼠模型，24 h后用XFZYD 治疗。4周后，超声心动图用于评估大鼠心功能。苏木素-伊红（HE）染色和Masson染色评估心肌组织病理学改变。将XFZYD灌胃SD大鼠（连续3天，每天一次），提取大鼠含药血清。分离培养原代新生大鼠心肌成纤维细胞（NRCFs），使用转化生长因子1β（TGF-1β）诱导48 h构建细胞模型。构建Janus激酶2（Janus kinase 2，JAK2）过表达（oe-JAK2）的NRCFs，用TGF-1β和含药血清处理48 h。Transwell迁移实验评估细胞迁移能力。使用MitoSOX Red和JC-1荧光探针分别检测线粒体活性氧（mtROS）和线粒体膜电位（MMP）。蛋白印迹法用于检测PTEN诱导假定激酶1（PINK1）、Parkin RBR E3泛素蛋白连接酶（Parkin）、微管相关蛋白1A/1B轻链3 （LC3）、α平滑肌肌动蛋白（α-SMA）、胶原（Collagen）I、Collagen III，以及JAK2/信号转导及转录激活因子3（STAT3）信号通路活性。结果：与假手术组比较，模型组大鼠左心室射血分数（LVEF）［（41.95±4.51)比（71.92±4.28），P<0.01］和左心室短轴缩短率（LVFS）［（16.08±0.97)比（35.75±1.93），P<0.01］下降，左心室舒张末期内径（LVEDD）和［（5.035±0.305）比（3.337±0.237），P<0.01］和左心室收缩末期内径（LVESD）［（9.108±0.171）比（6.855±0.261），P<0.01］提高，心肌损伤并伴有纤维化。与模型组比较，XFZYD组的上述异常指标均有显著改善。与Control组比较，TGF-β1组细胞迁移能力增强，α-SMA［（2.820±0.185）比（1.000±0.081），P<0.01］、Collagen I［（3.669±0.107）比（1.000±0.138），P<0.01］和Collagen III［（3.750±0.251）比（1.000±0.165），P<0.01］蛋白表达增加，mtROS含量增加、MMP下降，同时PINK1［（0.331±0.057）比（1.000±0.038），P<0.01］、Parkin［（0.442±0.073）比（1.000±0.095），P<0.01］蛋白表达和LC3Ⅱ/LC3Ⅰ比值［（0.168±0.036）比（1.000±0.127），P<0.01］显著减少，同时p-JAK2［（2.787±0.092）比（1.000±0.040)，P<0.01］和p-STAT3［（2.015±0.150)比（1.000±0.064），P<0.01］表达显著提高。与TGF-β1+NG组比较，TGF-β1+XFZYD组细胞迁移能力下降，且α-SMA［（1.629±0.095）比（2.827±0.147），P<0.01］、Collagen I［（1.629±0.115）比（3.530±0.128），P<0.01］和Collagen III［（1.592±0.066）比（3.791±0.260），P<0.01］蛋白表达减少，mtROS含量下降、MMP增加，且PINK1［（0.589±0.026）比(0.303±0.017），P<0.01］、Parkin［（0.683±0.017）比(0.405±0.020），P<0.01］蛋白表达和LC3Ⅱ/LC3Ⅰ比值［（0.692±0.104）比（0.161±0.016），P<0.01］明显提高，同时p-JAK2［（2.006±0.102）比（2.765±0.157），P<0.01］和p-STAT3［（1.500±0.163）比（2.102±0.153），P<0.01］蛋白表达减少。相对于TGF-β1+XFZYD+oe-NC组，TGF-β1+XFZYD+oe-JAK2组细胞JAK2、α-SMA［（0.947±0.125）比（0.531±0.017），P<0.01］、Collagen I［（0.644±0.070）比（0.370±0.026），P<0.01］和Collagen III［（0.553±0.043）比（0.277±0.022），P<0.01］蛋白表达增加，且PINK1［（1.433±0.133）比（3.211±0.165），P<0.01］、Parkin［（1.757±0.206）比（3.050±0.154），P<0.01］蛋白表达和LC3Ⅱ/LC3Ⅰ比值［（1.985±0.160）比（6.640±1.183），P<0.01］显著减少。结论：XFZYD可以通过抑制JAK/STAT信号通路调节线粒体自噬，从而改善MI后心室重构。

英文摘要:

Objective: To investigate the mechanism by which Xuefu Zhuyu Decoction (XFZYD) regulates mitochondrial autophagy to improve ventricular remodeling after myocardial infarction (MI) through in vivo and in vitro experiments. Methods: SD rats were divided into three groups: sham operation group, model group, and XFZYD group. An MI rat model was established by ligating the left anterior descending coronary artery, followed by XFZYD treatment 24 h later. After 4 weeks, echocardiography was used to assess cardiac function. Hematoxylin–eosin (HE) and Masson staining were applied to evaluate myocardial histopathological changes. Sprague–Dawley rats were administered XFZYD by gavage for 3 consecutive days (once daily), and drug-containing serum was prepared. Primary neonatal rat cardiac fibroblasts (NRCFs) were isolated and cultured, and a cellular model was established by treating NRCFs with transforming growth factor-β1 (TGF-β1) for 48 h. NRCFs with overexpressed Janus kinase 2 (oe-JAK2) were treated with TGF-β1 and drug-containing serum for 48 h. Cells were divided into 6 groups according to different treatment schemes: Control group, TGF-β1 group, TGF-β1+NG group, TGF-β1+XFZYD group, TGF-β1+XFZYD+oe-NC group, and TGF-β1+XFZYD+oe-JAK2 group. Cell migration was assessed by Transwell assay. Mitochondrial reactive oxygen species (mtROS) and mitochondrial membrane potential (MMP) were detected using MitoSOX Red and JC-1 probes, respectively. Western blotting was used to detect PTEN-induced putative kinase 1 (PINK1), Parkin RBR E3 ubiquitin-protein ligase (Parkin), microtubule-associated protein 1A/1B-light chain 3 (LC3), α-smooth muscle actin (α-SMA), collagen I, collagen III, as well as activation of the JAK2/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Results: Compared with the sham group, model group showed reduced LVEF [(41.95±4.51) vs. (71.92±4.28), P<0.01] and LVFS [(16.08±0.97) vs. (35.75±1.93), P<0.01], increased LVEDD [(5.035±0.305) vs. (3.337±0.237), P<0.01] and LVESD [(9.108±0.171) vs. (6.855±0.261), P<0.01], and myocardial injury with fibrosis. Compared with the model group, XFZYD significantly improved these abnormalities. In vitro, compared with the control group, TGF-β1 significantly enhanced cell migration, increased α-SMA [(2.820±0.185) vs. (1.000±0.081), P<0.01], Collagen I [(3.669±0.107) vs. (1.000±0.138), P<0.01], and Collagen III [(3.750±0.251) vs. (1.000±0.165), P<0.01] expression, elevated mtROS, and reduced MMP. PINK1 [(0.331±0.057) vs. (1.000±0.038), P<0.01], Parkin [(0.442±0.073) vs. (1.000±0.095), P<0.01] expression, and the LC3II/LC3I ratio [(0.168±0.036) vs. (1.000±0.127), P<0.01] were decreased, while p-JAK2 [(2.787±0.092) vs. (1.000±0.040), P<0.01] and p-STAT3 [(2.015±0.150) vs. (1.000±0.064), P<0.01] levels were elevated. Compared with the TGF-β1+NG group, the TGF-β1+XFZYD group showed reduced migration, downregulation of α-SMA [(1.629±0.095) vs. (2.827±0.147), P<0.01], Collagen I [(1.629±0.115) vs. (3.530±0.128), P<0.01], and Collagen III [(1.592±0.066) vs. (3.791±0.260), P<0.01], decreased mtROS, increased MMP, elevated PINK1 [(0.589±0.026) vs. (0.303±0.017), P<0.01], Parkin [(0.683±0.017) vs. (0.405±0.020), P<0.01], and LC3II/LC3I ratio [(0.692±0.104) vs. (0.161±0.016)，P<0.01], and reduced p-JAK2 [(2.006±0.102) vs. (2.765±0.157), P<0.01] and p-STAT3 [(1.500±0.163) vs. (2.102±0.153), P<0.01] expression. Compared with the TGF-β1+XFZYD+oe-NC group, cells in the TGF-β1+XFZYD+oe-JAK2 group exhibited significantly increased JAK2 expression, elevated α-SMA [(0.947±0.125) vs. (0.531±0.017), P<0.01], Collagen I [(0.644±0.070) vs. (0.370±0.026), P<0.01], and Collagen III [(0.553±0.043) vs. (0.277±0.022), P<0.01] levels, and decreased PINK1 [(1.433±0.133) vs. (3.211±0.165)，P<0.01], Parkin [(1.757±0.206) vs. (3.050±0.154), P<0.01], and LC3II/LC3I ratio [(1.985±0.160) vs. (6.640±1.183), P<0.01]. Conclusion: XFZYD improves ventricular remodeling after MI by regulating mitochondrial autophagy through inhibition of the JAK/STAT signaling pathway.

查看全文查看/发表评论下载PDF阅读器

关闭