基于AMPK/mTOR信号通路的白藜芦醇改善大鼠心力衰竭的作用机制

作者：韦晨龙，许锁春

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引用：引用：韦晨龙，许锁春.基于AMPK/mTOR信号通路的白藜芦醇改善大鼠心力衰竭的作用机制[J].中医药导报,2025,31(10):27-33,40.

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摘要：目的：探究白藜芦醇对心力衰竭模型大鼠的改善作用和对腺苷酸活化蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）通路的影响。方法：将6周龄的雄性SD大鼠随机分为假手术组（Sham）、模型组（Model）、白藜芦醇治疗组（RES）和白藜芦醇治疗加AMPK激动剂组（RES+EX229）。Sham组开胸后仅穿线不做冠状动脉左前支结扎，Model组给予冠状动脉左前支结扎，RES组在模型建立4周后，给予50 mg/kg的白藜芦醇灌胃8周，RES+EX229组在模型建立4周后，给予50mg/kg的白藜芦醇灌胃，并肌内注射EX229 2.0 mg/kg，共8周。多普勒超声仪测定心室功能指标，包括左心室舒张末期容积（LVEDV）、左心室收缩末期容积（LVESV）、左心室射血分数（LVEF）、左心室内径值（LV）、左室后壁（LVPW），以及心室重构指数，包括左心室质量指数（LVMI）和球形指数（SI）；TTC染色观察梗死面积，HE染色观察心脏组织病理学改变，Masson染色观察心肌胶原纤维变化，TUNEL染色观察心肌细胞凋亡情况，免疫组织化学染色观察心肌细胞自噬蛋白LC3的表达；Western blotting法对心脏组织B细胞淋巴瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）、微管相关蛋白1轻链3-Ⅱ型（LC3-Ⅱ）/LC3-Ⅰ、Beclin1、AMPK、mTOR蛋白表达情况进行测定。结果：与Sham组相比，Model组心脏的LVEDV、LVESV、LV、LVPW、LVMI升高，LVEF、SI降低，差异均有统计学意义（P＜0.05），与Model组相比，RES组LVEDV、LVESV、LV、LVPW、LVMI降低，LVEF、SI升高，差异均有统计学意义（P＜0.05），与RES组相比，RES+EX229组LVEDV、LVESV、LV、LVPW、LVMI升高，LVEF、SI降低，差异均有统计学意义（P＜0.05）；与Sham组相比，Model组的心肌梗死面积、病理程度、心肌胶原纤维程度、心肌细胞凋亡数量、自噬蛋白LC3和Beclin-1的表达明显上升，差异均有统计学意义（P＜0.05）；与Model组相比，RES组心肌梗死面积、病理程度、心肌胶原纤维程度、心肌细胞凋亡数量、自噬蛋白LC3的表达明显降低，差异均有统计学意义（P＜0.05）；与RES组相比，RES+EX229组心肌梗死面积，病理程度、心肌胶原纤维程度、心肌细胞凋亡数量、自噬蛋白LC3和Beclin1的表达明显升高，差异均有统计学意义（P＜0.05）；与Sham组相比，Model组的Bax、LC3-Ⅱ/LC3-Ⅰ、Beclin1、p-AMPK/AMPK表达量升高，Bcl-2、p-mTOR/mTOR表达量降低，差异均有统计学意义（P＜0.05）；与Model组相比，RES组的Bax、LC3-Ⅱ/LC3-Ⅰ、Beclin1、AMPK蛋白表达量升降低，Bcl-2、p-mTOR/mTOR表达量升高，差异均有统计学意义（P＜0.05）；与RES组相比，RES+EX229组Bax、LC3-Ⅱ/LC3-Ⅰ、Beclin1、p-AMPK/AMPK表达量升高，Bcl-2、p-mTOR/mTOR表达量降低，差异均有统计学意义（P＜0.05）。结论：白藜芦能够改善心力衰竭模型大鼠的心室功能、心脏组织病理学改变，抑制心肌细胞凋亡，改善心室重构，可能与白藜芦醇抑制AMPK/mTOR自噬信号通路激活有关。

关键词：白藜芦醇；心力衰竭；AMPK/mTOR信号通路；凋亡；自噬；大鼠

Abstract：Objective: To explore the improvement effect of resveratrol on heart failure model rat and its influence on AMPK/mTOR pathway. Method: The 6-week-old male SD rats were randomly divided into sham operation group (Sham group), model group (Model group), resveratrol treatment group (RES group), and resveratrol treatment plus AMPK agonist group (RES+EX229 group). In the Sham group, the chest was opened but no suture was made, and the left anterior descending coronary artery was not ligated. In the Model group, ligation of the left anterior descending coronary artery was performed. In the RES group, 50 mg/kg resveratrol was administered via gastric tube for 8 weeks after the model was established 4 weeks earlier. In the RES+EX229 group, 50 mg/kg resveratrol was administered via gastric tube after the model was established 4 weeks earlier, and 2.0 mg/kg EX229 was injected intramuscularly for 8 weeks. The left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular ejection fraction (LVEF), left ventricular inner diameter (LV), left ventricular posterior wall (LVPW), left ventricular remodeling index (LVMI) and spherical index (SI) were measured by Doppler ultrasound. TTC staining was used to observe the infarction area, and HE staining was used to observe heart tissue pathology change. Masson staining was used to observe changes of myocardial collagen fibers, and TUNEL staining was used to observe the myocardial cell apoptosis. Immunohistochemical staining was used to observe the myocardial cell autophagy genes expression of LC3, and Western blotting was used to detect expression of Bcl-2, Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1, AMPK and mTOR in cardiac tissue. Results: Compared with the Sham group, the LVEDV, LVESV, LV, LVPW and LVMI increased in Model group, while LVEF and SI decreased in Model group, with statistically significant difference (P<0.05). Compared with the Model group, the LVEDV, LVESV, LV, LVPW and LVMI decreased in RES group, while LVEF and SI increased in RES group, with statistically significant difference (P<0.05). Compared with  the RES group, the LVEDV, LVESV, LV, LVPW and LVMI increased in RES+EX229 group, while LVEF and SI decreased in RES+EX229 group, with statistically significant difference (P<0.05). Compared with the Sham group,  the myocardial infarction area, degree of pathology, degree of myocardial collagen fibers, number of myocardial cell apoptosis, and expression of LC3 and Beclin 1 increased in Model group, with statistically significant difference (P<0.05). Compared with the Model group, the myocardial infarction area, degree of pathology, degree of myocardial collagen fibers, number of myocardial cell apoptosis, and expression of LC3 decreased in RES group, with statistically significant difference (P<0.05). Compared with the RES group, the myocardial infarction area, degree of pathology, degree of myocardial collagen fibers, number of myocardial cell apoptosis, and expression of LC3 and Beclin-1 increased in RES+EX229 group, with statistically significant difference (P<0.05). Compared with the Sham group, the expression levels of Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1 and p-AMPK/AMPK increased in Model group, while Bcl-2 and p-mTOR/mTOR decreased in Model group, with statistically significant difference (P<0.05). Compared with the Model group, the expression levels of Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1 and AMPK decreased in RES group, while Bcl-2 and p-mTOR/mTOR increased in RES group, with statistically significant difference (P<0.05). Compared with the RES group, the expression levels of Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1 and p-AMPK/AMPK increased in RES+EX229 group, while Bcl-2 and p-mTOR/mTOR decreased in RES+EX229 group, with statistically significant difference (P<0.05). Conclusion: Resveratrol can improve ventricular function and pathological changes in cardiac tissue, inhibit myocardial cell apoptosis, and improve ventricular remodeling in heart failure model rats, which may be related to the inhibition of AMPK/mTOR autophagy signaling pathway activation.Objective: To explore the improvement effect of resveratrol on heart failure model rat and its influence on AMPK/mTOR pathway. Method: The 6-week-old male SD rats were randomly divided into sham operation group (Sham group), model group (Model group), resveratrol treatment group (RES group), and resveratrol treatment plus AMPK agonist group (RES+EX229 group). In the Sham group, the chest was opened but no suture was made, and the left anterior descending coronary artery was not ligated. In the Model group, ligation of the left anterior descending coronary artery was performed. In the RES group, 50 mg/kg resveratrol was administered via gastric tube for 8 weeks after the model was established 4 weeks earlier. In the RES+EX229 group, 50 mg/kg resveratrol was administered via gastric tube after the model was established 4 weeks earlier, and 2.0 mg/kg EX229 was injected intramuscularly for 8 weeks. The left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular ejection fraction (LVEF), left ventricular inner diameter (LV), left ventricular posterior wall (LVPW), left ventricular remodeling index (LVMI) and spherical index (SI) were measured by Doppler ultrasound. TTC staining was used to observe the infarction area, and HE staining was used to observe heart tissue pathology change. Masson staining was used to observe changes of myocardial collagen fibers, and TUNEL staining was used to observe the myocardial cell apoptosis. Immunohistochemical staining was used to observe the myocardial cell autophagy genes expression of LC3, and Western blotting was used to detect expression of Bcl-2, Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1, AMPK and mTOR in cardiac tissue. Results: Compared with the Sham group, the LVEDV, LVESV, LV, LVPW and LVMI increased in Model group, while LVEF and SI decreased in Model group, with statistically significant difference (P<0.05). Compared with the Model group, the LVEDV, LVESV, LV, LVPW and LVMI decreased in RES group, while LVEF and SI increased in RES group, with statistically significant difference (P<0.05). Compared with  the RES group, the LVEDV, LVESV, LV, LVPW and LVMI increased in RES+EX229 group, while LVEF and SI decreased in RES+EX229 group, with statistically significant difference (P<0.05). Compared with the Sham group,  the myocardial infarction area, degree of pathology, degree of myocardial collagen fibers, number of myocardial cell apoptosis, and expression of LC3 and Beclin 1 increased in Model group, with statistically significant difference (P<0.05). Compared with the Model group, the myocardial infarction area, degree of pathology, degree of myocardial collagen fibers, number of myocardial cell apoptosis, and expression of LC3 decreased in RES group, with statistically significant difference (P<0.05). Compared with the RES group, the myocardial infarction area, degree of pathology, degree of myocardial collagen fibers, number of myocardial cell apoptosis, and expression of LC3 and Beclin-1 increased in RES+EX229 group, with statistically significant difference (P<0.05). Compared with the Sham group, the expression levels of Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1 and p-AMPK/AMPK increased in Model group, while Bcl-2 and p-mTOR/mTOR decreased in Model group, with statistically significant difference (P<0.05). Compared with the Model group, the expression levels of Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1 and AMPK decreased in RES group, while Bcl-2 and p-mTOR/mTOR increased in RES group, with statistically significant difference (P<0.05). Compared with the RES group, the expression levels of Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1 and p-AMPK/AMPK increased in RES+EX229 group, while Bcl-2 and p-mTOR/mTOR decreased in RES+EX229 group, with statistically significant difference (P<0.05). Conclusion: Resveratrol can improve ventricular function and pathological changes in cardiac tissue, inhibit myocardial cell apoptosis, and improve ventricular remodeling in heart failure model rats, which may be related to the inhibition of AMPK/mTOR autophagy signaling pathway activation.Objective: To explore the improvement effect of resveratrol on heart failure model rat and its influence on AMPK/mTOR pathway. Method: The 6-week-old male SD rats were randomly divided into sham operation group (Sham group), model group (Model group), resveratrol treatment group (RES group), and resveratrol treatment plus AMPK agonist group (RES+EX229 group). In the Sham group, the chest was opened but no suture was made, and the left anterior descending coronary artery was not ligated. In the Model group, ligation of the left anterior descending coronary artery was performed. In the RES group, 50 mg/kg resveratrol was administered via gastric tube for 8 weeks after the model was established 4 weeks earlier. In the RES+EX229 group, 50 mg/kg resveratrol was administered via gastric tube after the model was established 4 weeks earlier, and 2.0 mg/kg EX229 was injected intramuscularly for 8 weeks. The left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular ejection fraction (LVEF), left ventricular inner diameter (LV), left ventricular posterior wall (LVPW), left ventricular remodeling index (LVMI) and spherical index (SI) were measured by Doppler ultrasound. TTC staining was used to observe the infarction area, and HE staining was used to observe heart tissue pathology change. Masson staining was used to observe changes of myocardial collagen fibers, and TUNEL staining was used to observe the myocardial cell apoptosis. Immunohistochemical staining was used to observe the myocardial cell autophagy genes expression of LC3, and Western blotting was used to detect expression of Bcl-2, Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1, AMPK and mTOR in cardiac tissue. Results: Compared with the Sham group, the LVEDV, LVESV, LV, LVPW and LVMI increased in Model group, while LVEF and SI decreased in Model group, with statistically significant difference (P<0.05). Compared with the Model group, the LVEDV, LVESV, LV, LVPW and LVMI decreased in RES group, while LVEF and SI increased in RES group, with statistically significant difference (P<0.05). Compared with  the RES group, the LVEDV, LVESV, LV, LVPW and LVMI increased in RES+EX229 group, while LVEF and SI decreased in RES+EX229 group, with statistically significant difference (P<0.05). Compared with the Sham group,  the myocardial infarction area, degree of pathology, degree of myocardial collagen fibers, number of myocardial cell apoptosis, and expression of LC3 and Beclin 1 increased in Model group, with statistically significant difference (P<0.05). Compared with the Model group, the myocardial infarction area, degree of pathology, degree of myocardial collagen fibers, number of myocardial cell apoptosis, and expression of LC3 decreased in RES group, with statistically significant difference (P<0.05). Compared with the RES group, the myocardial infarction area, degree of pathology, degree of myocardial collagen fibers, number of myocardial cell apoptosis, and expression of LC3 and Beclin-1 increased in RES+EX229 group, with statistically significant difference (P<0.05). Compared with the Sham group, the expression levels of Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1 and p-AMPK/AMPK increased in Model group, while Bcl-2 and p-mTOR/mTOR decreased in Model group, with statistically significant difference (P<0.05). Compared with the Model group, the expression levels of Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1 and AMPK decreased in RES group, while Bcl-2 and p-mTOR/mTOR increased in RES group, with statistically significant difference (P<0.05). Compared with the RES group, the expression levels of Bax, LC3-Ⅱ/LC3-Ⅰ, Beclin1 and p-AMPK/AMPK increased in RES+EX229 group, while Bcl-2 and p-mTOR/mTOR decreased in RES+EX229 group, with statistically significant difference (P<0.05). Conclusion: Resveratrol can improve ventricular function and pathological changes in cardiac tissue, inhibit myocardial cell apoptosis, and improve ventricular remodeling in heart failure model rats, which may be related to the inhibition of AMPK/mTOR autophagy signaling pathway activation.

Key words：resveratrol; heart failure; AMPK/mTOR signaling pathway; apoptosis; autophagy; rat

发布时间：2025-11-12

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