Vascular endothelial senescence induced by high glucose and palmitate (HG/PA) contributes to endothelial dysfunction, which leads to diabetic cardiovascular complications. Reduction of endothelial senescence may attenuate these pathogenic processes. This study is aimed at determining whether Ginseng-Sanqi-Chuanxiong (GSC) extracts, traditional Chinese medicine, can ameliorate human aortic endothelial cell (HAEC) senescence under HG/PA-stressed conditions and further explore the underlying mechanism. We found that GSC extracts significantly increased antisenescent activity by reducing the HG/PA-induced mitochondrial ROS (mtROS) levels in senescent HAECs. GSC extracts also induced cellular mitophagy formation, which mediated the effect of GSC extracts on mtROS reduction. Apart from this, the data showed that GSC extracts stimulated mitophagy via the AMPK pathway, and upon inhibition of AMPK by pharmacological and genetic inhibitors, GSC extract-mediated mitophagy was abolished which further led to reverse the antisenescence effect. Taken together, these data suggest that GSC extracts prevent HG/PA-induced endothelial senescence and mtROS production by mitophagy regulation via the AMPK pathway. Thus, the induction of mitophagy by GSC extracts may provide a novel therapeutic candidate for cardiovascular protection in metabolic syndrome.

Shenfu Tang and Dushen Tang (one of the composite medicines for Shenfu Tang) are widely used Traditional Chinese herbal formulations and ginsenosides are their main bioactive components. However, there are rare studies about simultaneous analysis of ginsenosides in Shenfu Tang and Dushen Tang. In order to identify ginsenosides in Shenfu Tang and Dushen Tang and to explore law of compatibility of medicines in the decoction, a method for simultaneous determination of twelve ginsenosides in Shenfu Tang and Dushen Tang was developed by ultraresolution liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). The method showed satisfactory linearity (r > 0.9915), repeatability (RSD < 9.58%), intra- and interday precisions (RSD<11.90%), and high yields of recovery (92.26-113.20%) for twelve major constituents, namely, ginsenosides-Rb1, Rb2, Rb3, Rc, Rd, Rg1, Re, Rf, Rg2, Rg3, Rh1, and F2. Furthermore, the concentration of twelve ginsenosides in Dushen Tang and Shenfu Tang was also simultaneously analyzed. Most of ginsenosides except Rg1 and Rb1 showed higher contents in Shenfu Tang compared to Dushen Tang. The compatibility of the formula had the effect of promoting or inhibiting the dissolution of some major components. The present research provided a reliable evidence for the illustration of chemical basis and compatibility regularity of Shenfu Tang. This study demonstrated the utility of the developed method for assessment of the quantity of the major constituents in Dushen Tang and Shenfu Tang.

Vascular aging is an independent risk factor for age-related diseases and a specific type of organic aging. Endothelial progenitor cells (EPCs), a type of bone marrow stem cell, has been linked to vascular aging. The purpose of this study is to investigate if Ginseng-Sanqi-Chuanxiong (GSC) extract, a traditional Chinese medicine, can delay aortic aging in mice by enhancing the performance and aging of EPCs in vivo and to analyze the potential mechanisms through a d-Galactose (D-gal)-induced vascular aging model in mice. Our study revealed that GSC extracts not only enhanced the aortic structure, endothelial function, oxidative stress levels, and aging in mice, but also enhanced the proliferation, migration, adhesion, and secretion of EPCs in vivo, while reducing the expression of p53, p21, and p16. To conclude, GSC can delay vascular senescence by enhancing the function and aging of EPCs, which could be linked to a decrease in p16 and p53/p21 signaling. Consequently, utilizing GSC extracts to enhance the function and senescence of autologous EPCs may present a novel avenue for enhancing autologous stem cells in alleviating senescence.

Background: During the aging process, the number and functional activity of endothelial progenitor cells (EPCs) are impaired, leading to the unsatisfactory efficacy of transplantation. Previous studies demonstrated that Yiqihuoxue decoction (Ginseng-Sanqi-Chuanxiong, GSC) exerts anti-vascular aging effects. The purpose of this study is to evaluated the effects of GSC on D-galactose (D-gal)induced senescence and the underlying mechanisms. Methods: The levels of cellular senescence-related markers P16, P21, P53, AMPK and p-AMPK were detected by Western blot analysis (WB). SA-β-gal staining was used to evaluate cell senescence. EPCs function was measured by CCK-8, Transwell cell migration and cell adhesion assay. The morphological changes of mitochondria were detected by confocal microscopy. The protein and mRNA expression of mitochondrial fusion fission Drp1, Mff, Fis1, Mfn1, Mfn2 and Opa1 in mitochondria were detect using WB and RT–qPCR. Mitochondrial membrane potential, mtROS and ATP of EPCs were measured using IF. H&E staining was used to observe the pathological changes and IMT of the aorta. The expressions of AGEs, MMP-2 and VEGF in aorta were measured using Immunohistochemical (IHC). The levels of SOD, MDA, NO and ET-1 in serum were detected by SOD, MDA and NO kits. Results: In vitro, GSC ameliorated the senescence of EPCs induced by D-gal and reduced the expression of P16, P21 and P53. The mitochondrial morphology of EPCs was restored, the expression of mitochondrial Drp1, Mff and Fis1 protein was decreased, the levels of mtROS and ATP were decreased, and mitochondrial function was improved. Meanwhile, the expression of AMPK and p-AMPK increased. The improvement effects of GSC on aging and mitochondrial morphology and function were were hindered after adding AMPK inhibitor. In vivo, GSC improved EPCs efficiency, ameliorated aortic structural disorder and decreased IMT in aging mice. The serum SOD level increased and MDA level decreased, indicating the improvement of antioxidant capacity. Increased NO content and ET-1 content suggested improvement of vascular endothelial function. The changes observed in SOD and MMP-2 suggested a reduction in vascular stiffness and the degree of vascular damage. The decreased expression of P21 and P53 indicates the delay of vascular senescence.

Objective: To investigating the molecular mechanism of herbal pairs in three types of Chinese medicinals: Qi-tonifying, blood activation, blood-stasis breaking in treatment of coronary heart disease (CHD). Methods: The components of six herbs were searched in Chinese medicine dictionary and their target proteins were found in PubChem. CHD genes were obtained from PubMed gene database. Ingenuity Pathways Analysis was used to build the pharmacological network of three herbal pairs and CHD molecular network. The canonical pathways between each herbal pair network and CHD network was compared to decipher the molecular mechanism on three herbal pairs in treating CHD. Results: The network analysis showed that there were the common signal pathways of three herbal pairs in treating CHD including hypoxia signaling in the cardiovascular system, Hypoxia-inducible factor 1-alpha signaling, glucocorticoid receptor signaling, G-Protein coupled receptor signaling and pregnane X receptor/retinoid X receptor (PXR/RXR) activation. Further to analyze cardiovascular signaling, cytokine signaling and cytokine signaling, the effective molecules for three herbal pairs in treating CHD included HIF1α and estrogen receptor 1, Qi-tonifying herbal pair included albumin and matrix metallopeptidase 2, and blood-activation herbal pair included estrogen receptor 2 and peroxisome proliferator-activated receptor-γ. Conclusion: Each herbal pair can affect some respective CHD-related functions and pathways, meanwhile three herbal pairs exert some mutual effects on CHD-related functions and pathways. Mutual effects of three herbal pairs may be the key components of their total molecular mechanisms and respective effects of each herbal pair may be the characteristic components of their respective molecular mechanism.