Department of Pharmacology, School of Basic Medical Sciences, Anhui Medical University, No.81 Meishan Road, Hefei, 230032, PR China; Department of School of Pharmacy, Chongqing Health Vocational College, NO.99 Xirong Road, Dazu, 404100, PR China.
Department of Pharmacology, School of Basic Medical Sciences, Anhui Medical University, No.81 Meishan Road, Hefei, 230032, PR China.
Phytomedicine. 2024 Jun;128:155319. doi: 10.1016/j.phymed.2023.155319. Epub 2023 Dec 25.
Myocardial infarction (MI) is one of the most deadly diseases in the world. Hyperoside (Hyp) has been shown to have a protective effect on cardiovascular function through various signaling pathways, but whether it can protect myocardial infarction by regulating JAK2/STAT3 signaling pathway is unknown.
To investigate whether Hyp could protect the heart against myocardial infarction injury in mice by modulating JAK2/STAT3 signaling pathway and its potential mechanism.
In vivo experiments, the myocardial infarction model was established by ligating the left anterior descending coronary artery (LAD) of male C57BL/6 mice permanently. The mice were divided into seven groups: sham group, MI group, MI+Hyp (9 mg/kg), MI+Hyp (18 mg/kg) group, MI+Hyp (36 mg/kg) group, MI+Captopril group (15 mg/kg) group and MI+Hyp (36 mg/kg)+AG490 (7.5 mg/kg) group. Each group of animals were given different concentrations of hyperoside, positive control drug or inhibitor of JAK2/STAT3 singaling. After 14 days of administration, the electrocardiogram (ECG), echocardiography and serum myocardial injury markers were examined; Slices of mouse myocardial tissue were assessed for histopathological changes by HE, Masson and Sirius Red staining. TTC and TUNEL staining were used to evaluate the myocardial infarction area and cardiomyocytes apoptosis respectively. The expression of JAK2/STAT3 signaling pathway, apoptosis and autophagy-related proteins were detected by western blot. In vitro experiments, rat H9c2 cardiomyocytes were deprived of oxygen and glucose (OGD) to stimulate myocardial ischemia. The experiment was divided into seven groups: Control group, OGD group, OGD+Hyp (20 μM) group, OGD+Hyp (40 μM) group, OGD+Hyp (80 μM), OGD+Captopril (10 μM) group and OGD+Hyp (80 μM)+AG490 (100 μM) group. Myocardial cell damage and redox index were measured 12 h after OGD treatment. ROS content in cardiomyocytes was detected by immunofluorescence. Cardiomyocytes apoptosis was detected by flow cytometry. The expressions of JAK2/STAT3 signaling pathway-related proteins, apoptosis and autophagy related proteins were detected by western blot.
In vivo, hyperoside could ameolirate ECG abnormality, increase cardiac function, reduce myocardial infarction size and significantly reduce myocardial fibrosis level and oxidation level. The experimental results in vitro showed that Hyp could reduce the ROS content in cardiomyocytes, decrease the level of oxidative stress and counteract the apoptosis induced by OGD injury . Both in vivo and in vitro experiments showed that hyperoside could increase phosphorylated JAK2 and STAT3, indicating that hyperoside could play a cardioprotective role by activating JAK2/STAT3 signaling pathway. It was also shown that hyperoside could increase the autophagy level of cardiomyocytes in vivo and in vitro. However the cardiomyocyte-protective effect of Hyp was abolished in combination with JAK2/ STAT3 signaling pathway inhibitor AG490. These results indicated that the protective effect of Hyp on cardiomyocyte injury was at least partially achieved through the activation of the JAK2/STAT3 signaling pathway.
Hyp can significantly improve cardiac function, ameliorate myocardial hypertrophy and myocardial remodeling in MI mice. The mechanism may be related to improving mitochondrial autophagy of cardiomyocytes to maintain the advantage of autophagy, and blocking apoptosis pathway through phagocytosis, thus suppressing apoptosis level of cardiomyocytes. These effects of Hyp are achieved, at least in part, by activating the JAK2/STAT3 signaling pathway.
心肌梗死（MI）是世界上最致命的疾病之一。已经表明，金丝桃苷（Hyp）通过各种信号通路对心血管功能具有保护作用，但它是否可以通过调节 JAK2/STAT3 信号通路来保护心肌梗死尚不清楚。
研究 Hyp 是否可以通过调节 JAK2/STAT3 信号通路及其潜在机制来保护小鼠心肌梗死损伤。
体内实验中，通过结扎雄性 C57BL/6 小鼠的左前降支冠状动脉（LAD）永久性建立心肌梗死模型。将小鼠分为七组：假手术组、MI 组、MI+Hyp（9mg/kg）组、MI+Hyp（18mg/kg）组、MI+Hyp（36mg/kg）组、MI+Captopril 组（15mg/kg）组和 MI+Hyp（36mg/kg）+AG490（7.5mg/kg）组。每组动物给予不同浓度的金丝桃苷、阳性对照药物或 JAK2/STAT3 信号通路抑制剂。给药 14 天后，检查心电图（ECG）、超声心动图和血清心肌损伤标志物；通过 HE、Masson 和 Sirius Red 染色评估小鼠心肌组织切片的组织学变化。TTC 和 TUNEL 染色分别用于评估心肌梗死面积和心肌细胞凋亡。通过 Western blot 检测 JAK2/STAT3 信号通路、凋亡和自噬相关蛋白的表达。在体外实验中，用缺氧和葡萄糖剥夺（OGD）刺激大鼠 H9c2 心肌细胞建立心肌缺血模型。实验分为七组：对照组、OGD 组、OGD+Hyp（20μM）组、OGD+Hyp（40μM）组、OGD+Hyp（80μM）组、OGD+Captopril（10μM）组和 OGD+Hyp（80μM）+AG490（100μM）组。OGD 处理后 12h 测量心肌细胞损伤和氧化还原指数。通过免疫荧光法检测心肌细胞内 ROS 含量。通过流式细胞术检测心肌细胞凋亡。通过 Western blot 检测 JAK2/STAT3 信号通路相关蛋白、凋亡和自噬相关蛋白的表达。
体内实验中，金丝桃苷可改善 ECG 异常、增加心功能、减少心肌梗死面积，显著降低心肌纤维化水平和氧化水平。体外实验结果表明，Hyp 可降低心肌细胞内 ROS 含量，降低氧化应激水平，并拮抗 OGD 损伤诱导的细胞凋亡。体内和体外实验均表明，金丝桃苷可增加磷酸化 JAK2 和 STAT3 的表达，提示金丝桃苷通过激活 JAK2/STAT3 信号通路发挥心肌保护作用。还表明，金丝桃苷可增加体内和体外心肌细胞的自噬水平。然而，当与 JAK2/STAT3 信号通路抑制剂 AG490 联合使用时，Hyp 的心肌细胞保护作用被消除。这些结果表明，Hyp 对心肌细胞损伤的保护作用至少部分是通过激活 JAK2/STAT3 信号通路实现的。
Hyp 可显著改善 MI 小鼠的心脏功能，改善心肌肥厚和心肌重塑。其机制可能与改善心肌细胞线粒体自噬以维持自噬优势有关，并通过吞噬作用阻断凋亡途径，从而抑制心肌细胞的凋亡水平。Hyp 的这些作用至少部分是通过激活 JAK2/STAT3 信号通路来实现的。
