血栓通对糖尿病内皮细胞线粒体动态平衡、内皮功能损伤和动脉粥样硬化的保护作用研究

doi:10.12092/j.issn.1009-2501.2026.06.009

摘要/Abstract

摘要：

目的: 研究血栓通（XST）治疗糖尿病加速动脉粥样硬化（AS）的作用及机制。方法: 采用ApoE^?/?小鼠和C57BL/6J小鼠，通过腹腔注射链脲佐菌素（STZ）建立糖尿病AS模型，并给予XST干预12周。检测空腹血糖水平、血清总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）和高密度脂蛋白胆固醇（HDL-C）含量，通过苏丹Ⅳ及油红O染色观察主动脉弓和主动脉根部斑块及脂质沉积情况来评价AS，离体血管环实验评估主动脉内皮依赖性舒张功能，透射电镜观察主动脉内皮细胞中线粒体形态变化，采用高葡萄糖刺激人脐静脉内皮细胞（HUVECs），共聚焦显微镜下观察线粒体形态及其分裂、融合情况。同时检测线粒体活性氧（ROS）水平，并采用蛋白免疫印迹检测线粒体动态调控蛋白的表达。结果: XST对糖尿病AS模型小鼠血糖血脂无明显影响，XST显著减少糖尿病AS模型小鼠主动脉弓及根部的斑块面积，并改善STZ诱导的C57BL/6J小鼠中乙酰胆碱（ACh）介导的内皮依赖性舒张功能障碍。在高糖诱导HUVECs模型中，XST显著抑制线粒体碎片化，减少ROS产生。机制研究表明，高糖刺激可上调动力相关蛋白1（Drp1）和线粒体分裂蛋白1（Fis1）的表达并降低磷酸化腺苷酸活化蛋白激酶（p-AMPK）水平，而XST处理可显著上调p-AMPK表达，抑制Drp1和Fis1的表达。结论: XST可能通过激活AMPK/Drp1信号通路，调控内皮线粒体动态平衡，从而抑制糖尿病加速的动脉粥样硬化进程。

关键词: 血栓通, 糖尿病动脉粥样硬化, 内皮功能障碍, 线粒体动态平衡, 动力相关蛋白1

Abstract:

AIM: To investigate the therapeutic effect and underlying mechanisms of Xueshuantong (XST) in diabetes-accelerated atherosclerosis (AS). METHODS: Diabetes-accelerated AS models were established in ApoE^?/? mice and C57BL/6J mice by intraperitoneal injection of streptozotocin (STZ) followed by XST administration for 12 weeks. Fasting blood glucose and serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were measured. Atherosclerosis was assessed by evaluating plaque formation and lipid deposition in the aortic arch and root using Sudan IV and Oil Red O staining. Endothelium-dependent vasorelaxation was evaluated using isolated aortic ring assays. Mitochondrial morphology in aortic endothelial cells was examined by transmission electron microscopy. Human umbilical vein endothelial cells (HUVECs) were exposed to high glucose and treated with XST. Mitochondrial morphology and dynamics were analyzed using confocal microscopy. Mitochondrial reactive oxygen species (ROS) levels were measured, and the expression of mitochondrial dynamics-related protein was assessed by Western blotting. RESULTS: XST had no significant effect on blood glucose or lipid levels in diabetic AS mice. XST significantly reduced atherosclerotic plaque areas in the aortic arch and root of diabetic ApoE^?/^? mice, and improved acetylcholine-mediated endothelium-dependent vasorelaxation in STZ-induced C57BL/6J mice. In high glucose-stimulated HUVECs, XST significantly inhibited mitochondrial fragmentation and reduced mitochondrial ROS production. Mechanistically, high glucose increased the expression of dynamin-related protein 1 (Drp1) and mitochondrial fission protein 1 (Fis1), while decreasing phosphorylated AMP-activated protein kinase (p-AMPK) levels. XST treatment reversed these effects by activating AMPK and downregulating Drp1 and Fis1. CONCLUSION: XST may alleviates diabetes-accelerated atherosclerosis by activating the AMPK/Drp1 signaling pathway and restoring mitochondrial dynamic balance in endothelial cells.

Key words: Xueshuantong, diabetic atherosclerosis, endothelial dysfunction, mitochondrial dynamics, Drp1

中图分类号:

R965.2

祁光伟, 仝婉昱, 王冀鲁, 郭靖文, 黎瑞巧, 王启隆. 血栓通对糖尿病内皮细胞线粒体动态平衡、内皮功能损伤和动脉粥样硬化的保护作用研究[J]. 中国临床药理学与治疗学, 2026, 31(6): 798-807.

Guangwei QI, Wanyu TONG, Jilu WANG, Jingwen GUO, Ruiqiao LI, Qilong WANG. Protective effects of Xueshuantong on mitochondrial dynamic balance, endothelial dysfunction, and atherosclerosis in diabetes[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(6): 798-807.

图/表 6

图 1

Fig.1 Effects of XST on body weight, blood glucose and lipid profiles in diabetic ApoE?/? mice A: weight changes of ApoE?/? mice within 12 weeks. B: blood glucose changes of ApoE?/? mice within 12 weeks. C-F: the serum levels of TC, TG, HDL-C and LDL-C in STZ-induced diabetic ApoE?/? mice. n = 6-10 per group. bP < 0.05, compared with Con group; eP < 0.05, compared with MOD group.

图 2

Fig.2 XST attenuates atherosclerotic lesions in diabetic ApoE?/? mice A: representative images of atherosclerosis lesion at the aortic arch. B: representative images of Sudan IV staining of atherosclerotic lesions at the aortic arch. C: representative images of Oil Red O staining of atherosclerotic lesions in the aortic sinus. D: quantification of en face atherosclerotic lesion area in the aortic arch. E: quantitative evaluation of the size of atherosclerotic lesions in the aortic root. n = 6-10 per group. cP<0.01, compared with Con group; fP<0.01, compared with STZ group.

图 3

Fig.3 XST prevents endothelial dysfunction under diabetic conditions A-B: U46619 (10 nmol/L) was used to contract the aortic rings. A: endothelium-dependent vasodilator responses were measured by Ach (10?9 to 10?5 mol/L). B: endothelium-independent vasodilator responses were measured in the presence of SNP (10?10 to 10?6 mol/L). C: p-eNOS in the aorta, was analyzed by Western blot. D: quantitative analysis of p-eNOS. n = 3. bP< 0.05, compared with Con group, eP< 0.05, compared with STZ group. E: representative fluorescence images are shown. F: fluorescence intensity quantification for mitochondrial ROS levels in HUVECs. bP< 0.05, compared with NG group, eP< 0.05, fP< 0.01, compared with HG group.

图 4

Fig.4 XST prevents hyperglycemia-induced mitochondrial fragmentation in diabetic aortic endothelium A: representative electron microscopic images of mitochondria in the aortic endothelium (original magnification 25 000×, scale bar = 500 nm). B: quantification of mitochondrial length. At least 50 mitochondria per mouse were analyzed (n=3). C: mitochondrial dynamics-related proteins, including Drp1, Fis1, and p-AMPK in the aorta, were analyzed by Western blot. D-F: quantitative analysis of Drp1, Fis1 and p-AMPK. n=3, bP<0.05, cP<0.01, compared with Con group, eP<0.05, fP<0.01, compared with STZ group.

图 5

Fig.5 XST prevents high glucose-induced mitochondrial fission in HUVECs A: mitochondria were labeled with MitoTracker Deep Red, and mitochondrial morphology was analyzed using fluorescence microscopy (scale bar=20 μm). B: mitochondrial morphology in live HUVECs stained with MitoTracker Deep Red FM and captured using time-lapse confocal microscopy. Images were collected at 1 min intervals for 6 min. C-D: mitochondrial volume and number were quantified. E: quantitative analysis of mitochondrial fission events. n≥30 cells. bP<0.05, cP<0.01, compared with NG group, eP<0.05, fP<0.01, compared with HG group.

图 6

Fig.6 XST reduces Drp1 expression via AMPK activation A: mitochondrial dynamics-related proteins, including Drp1, Fis1, MFF and OPA1 were analyzed by Western blot. B-C: quantitative analysis of Drp1 and Fis1. D: p-AMPK was analyzed by Western blotting. E: quantitative analysis of p-AMPK. n=3, bP<0.05, cP<0.01 compared with NG group, eP<0.05, fP<0.01, compared with HG group.

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