Protective effects of Xueshuantong on mitochondrial dynamic balance, endothelial dysfunction, and atherosclerosis in diabetes

doi:10.12092/j.issn.1009-2501.2026.06.009

Abstract

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

CLC Number:

R965.2

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.

Figures/Tables 6

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.

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.

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.

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.

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.

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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