Objective:To observe the effect of Ginseng Radix et Rhizoma， Notoginseng Radix et Rhizoma， and Chuanxiong Rhizoma extract （GNC） on mitochondrial oxidative stress in hydrogen peroxide （H₂O₂）-induced aging of human umbilical vein endothelial cells （HUVECs）， and explore the therapeutic mechanism of GNC on aging HUVECs. Method:The HUVECs were classified into the control group （control）， H₂O₂ model group （H₂O₂）， H₂O₂ + DMSO group （DMSO， 1 mL·L^-1）， resveratrol group （Resv， 8 μmol·L^-1）， and low- （200 mg·L^-1）， medium- （300 mg·L^-1）， and high-dose （400 mg·L^-1） GNC （GNC-L， GNC-M， and GNC-H） groups. Except control group and H₂O₂ group， the other groups were intervened with corresponding agents. Subsequently， 300 μmol·L^-1 H₂O₂ was given to other groups except the control group for 4 h to induce aging， and then the cells were cultured in normal media for 24 h. The aging degree， cell cycle， and mitochondrial reactive oxygen species （mtROS） level were determined by SA-β-galactosidase （SA-β-Gal） staining， flow cytometry， and MitoSox red fluorescence staining， respectively. JC-10 was used as a fluorescent probe to detect the changes in mitochondrial membrane potential， and Western blot was performed to detect the expression of manganese superoxide dismutase （MnSOD） and p-p66 proteins. Result:The SA-β-gal staining results showed that H₂O₂ group had increased blue-stained cells compared with other groups （P<0.01）. Compared with those in the control group， the ratio of G₀/G₁ phase cells significantly increased （P<0.05） and that of G₂/M phase cells decreased （P<0.05） in the H₂O₂ group. Compared with those in the H₂O₂ group， the proportion of G₀/G₁ cells decreased （P<0.05） while that of G₂/M cells increased （P<0.05） in GNC-H groups and Resv group. The fluorescence staining for determining mitochondrial ROS level showed that the H₂O₂ group had weakened fluorescence intensity than the control， GNC-H， and GNC-M groups （P<0.05）. The mitochondrial membrane potential fluorescence intensity of the H₂O₂ group was weaker than that of the control， GNC-H， GNC-M， and GNC-L groups （P<0.01）， as well as the Resv group （P<0.05）. Western blot showed that the protein level of MnSOD was significantly lower in the H₂O₂ group than in the control， GNS-H， and GNS-M groups （P<0.05）， whereas the protein level of p-p66 showed an opposite trend （P<0.01）， indicating that the medication can alleviate the intracellular mitochondrial oxidative stress. Conclusion:GNC can delay the H₂O₂-induced aging of vascular endothelial cells. The GNC intervention significantly regulated the mitochondrial ROS， mitochondrial membrane potential， and related proteins MnSOD and p-p66 to alleviate oxidative stress. Chinese medicinal materials may delay the aging of vascular endothelial cells by inhibiting mitochondrial oxidative stress.

Objective:To investigate the effect of Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract on endothelial microparticles （EMPs）-induced vascular endothelial cell senescence， and explore the possible mechanism. Method:Human umbilical vein endothelial cells （HUVECs） were used as the research objects， and the aged model was established with 10-12 passages of replicative senescence cells. The experimental cells were divided into young group （2-4 passage cells）， aged group （10-12 passage cells）， only EMPs intervention group （extract EMPs produced by aged cells to intervene young cells） and low dose， middle dose and high dose drug intervention groups （200， 300， 400 mg·L^-1）. Senescence related β-galactosidase （SA-β-gal） staining and cell cycle propidium iodide （PI） staining were used to determine cell senescence. Cell counting kit-8 （CCK-8） assay was used to screen the drug concentration. EMPs were extracted by two-step centrifugation， EMPs labeled with phycoerythrin （PE） anti-human CD31 antibody or fluorescein isothiocyanate （FITC） annexin V were detected by flow cytometry， intracellular reactive oxygen species （ROS） were detected by 2'，7'- dichlorofluorescein diacetate （DCFDA） staining. Result:After treatment with the drug， SA-β-gal activity of the aged cells significantly decreased （P<0.01）， the S phase arrest was restored （P<0.01）， and the number of CD31⁺ EMPs and annexin V⁺ EMPs secreted by aged cells decreased （P<0.05）. Compared with the young group， only EMPs intervention group could induce increased SA-β-gal activity and S phase arrest in young cells （P<0.05，P<0.01）. However， after intervention of EMPs and the drug， EMPs-mediated increase of SA-β-gal activity was significantly inhibited and S phase arrest was restored （P<0.05）. The increase of intracellular ROS induced by EMPs was also significantly inhibited by the drug （P<0.05，P<0.01）. Conclusion:Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract can delay the senescence of vascular endothelial cells by influencing EMPs， and the mechanism may be related to the inhibition of increased intracellular ROS induced by EMPs.

Microarray data of hippocampal tissue(HC) of the cognitively intact elderly(60-99 years old) were compared with those of the middle-aged and the young(20-59 years old) by bioinformatics techniques to initially screen out differentially expressed genes(DEGs) and then predict potential effective Chinese medicinals for the treatment of brain aging. The gene expression profile(accession: GSE11882) was downloaded from the Gene Expression Omnibus(GEO) and DEGs were screened based on R package. The key DEGs were identified by STRING, Cytoscape and the plug-in, followed by Gene Ontology(GO) term and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis. Then the key genes and the medical ontology information retrieval platform(Coremine Medical) were mapped against each other to single out the Chinese medicinals for the treatment of brain aging and construct the " Chinese medicinal-active constituent-target" network. Among the resultant 268 DEGs(246 down-regulated and 22 up-regulated), the 15 key genes were mainly involved in biological processes such as leukocyte migration, neutrophil activation, cell chemotaxis, microglia activation and response to external stimulus, and pathways such as inflammatory process, immune response, cytokine-cytokine receptor interaction, PI3 K-Akt signaling pathway, Rap1 signaling pathway, chemokine signaling pathway and Toll-like receptor signaling pathway. The potential effective Chinese medicinals were Notoginseng Radix et Rhizoma, Ginseng Radix et Rhizoma, Salviae Miltiorrhizae Radix et Rhizoma and Astragali Radix. The analysis of DEGs and key genes enhances the understanding of the mechanisms of brain aging. This study provides potential gene targets and ideas for the development of Chinese medicine for brain aging.
Adult ; Aged ; Aged, 80 and over ; Brain ; China ; Computational Biology ; Gene Expression Profiling ; Gene Ontology ; Humans ; Middle Aged ; Transcriptome ; Young Adult

This study aims to explore the effect of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Chuanxiong Rhizoma(hereinafter referred to as GNS) on the SIRT1-autophagy pathway of endothelial cell senescence induced by hydrogen peroxide(H_2O_2). To be specific, vascular endothelial cells were classified into the blank control group(control), model group(model), model + DMSO group(DMSO), resveratrol group(RESV), and GNS low-dose(GNS-L), medium-dose(GNS-M), and high-dose(GNS-H) groups. They were treated with H_2O_2 for senescence induction except the control. After intervention of cells in each group with corresponding drugs for 24 h, cell growth status was observed under an inverted microscope, and the formation of autophagosome under the transmission electron microscope. In addition, the changes of microtubule-associated protein 1 light chain 3β(LC3 B) were detected by immunofluorescence staining. The autophagy flux was tracked with the autophagy double-labeled adenovirus(mRFP-GFP-LC3) fusion protein. Dansylcadaverine(MDC) staining was employed to determine the autophagic vesicles, and Western blot the expression of sirtuin 1(SIRT1), ubiquitin-binding protein p62, and LC3Ⅱ. After H_2O_2 induction, cells demonstrated slow growth, decreased adhesion ability, raised number of SA-β-gal-stained blue ones, a certain number of autophagosomes with bilayer membrane and secondary lysosomes in the cytoplasm, and slight rise of autophagy flux level. Compared with the model group, GNS groups showed improved morphology, moderate adhesion ability, complete and smooth membrane, decreased SA-β-gal-stained blue cells, many autophagosomes, autophagic vesicles, and secondary lysosomes in the cytoplasm, increased autophagolysosomes, autophagy flux level, and fluorescence intensity of LC3 B and MDC, up-regulated expression of SIRT1 and LC3Ⅱ, and down-regulated expression of p62, suggesting the improvement of autophagy level. GNS can delay the senescence of vascular endothelial cells. After the intervention, the autophagy flux and related proteins SIRT1, LC3Ⅱand p62 changed significantly, and the autophagy level increased significantly. However, EX527 weakened the effect of Chinese medicine in delaying vascular senescence. GNS may delay the senescence of vascular endothelial cells through the SIRT1 autophagy pathway.
Autophagy ; Cells, Cultured ; Cellular Senescence ; Drugs, Chinese Herbal/pharmacology* ; Endothelial Cells/drug effects* ; Hydrogen Peroxide ; Panax/chemistry* ; Sirtuin 1/genetics*

The application of clinical medication and basic research progress of traditional Chinese medicine （TCM） for Yiqi Huoxue Huatan in the treatment of atherosclerosis （AS） were summarized. According to the different pathogenic sites of AS， the clinical research progress of TCM for Yiqi Huoxue Huatan in the treatment of AS and the commonly used TCM for the treatment of AS were summarized. Astragali Radix， Salviae Miltiorrhizae Radix， Quinquefolium Panax， Cocos Wolf Poria， Atractylodis Rhizoma， Rosea Rhodiola， which were Yiqi herbs， were mostly used for the treatment of AS. Wallichii Ligusticum， Salviae Miltiorrhizae Radix， Notoginseng Radix， Paeoniae Rubra Radix， Paeoniae Alba Radix， Angelicae Sinensis Radix， Semen Persicae， Tinctorius Carthamus， Achyranthis Bidentatae Radix， tea root， which were Huoxue herbs， were mostly used for the treatment of AS. Huatan herbs， including Kirilowii Maxim Trichosanthes， Pinelliae Rhizama， Acorus Tatarinowii Schott， Citri Reticulatae Pericarpium， Cum Bile Arisaema， Silicea Bambusae Concretio， Aurantii Immaturus Fructus， Bamboo Juice， were commonly used for the treatment of AS. According to the findings， TCM for Yiqi Huoxue was mostly combined with insect medicine and rattan medicine for the treatment of carotid atherosclerosis， combined with TCM for promoting Qi， relieving pain， dissipating blood stasis and reducing phlegm for the treatment of coronary heart disease， and combined with TCM for relaxing tendons and activating collaterals， resolving phlegm to benefit orifices， and invigorating spleen to remove dampness combined for the treatment of lower extremity sclerosis. In addition， the medication time， drug combination and improvement indexes were summarized. In basic studies， the experimental progress of this kind of medicine for the treatment of AS were summed up in the aspect of reducing inflammatory reaction， improving the abnormal lipid metabolism and improving the damage of inner membrane. At present， it was found that tanshinone， total saponins of stem and leaf of Panax Quinquefolium， extract of Trichosanthis Pericarpium. Qishen Yiqi dropping pill， Huxinkang tablet， Danlou tablet， Buyang Huanwutang combined with Gualou Xiebaitang， Huazhuo Tongmai powder were the main drugs for basic research， and the animal model， model characteristics and the mechanism of action were summarized. In order to provide a reference for the rational application of TCM for Yiqi Huoxue Huatan in the treatment of AS， the application law， the mechanism and characteristics of action and the future research directions of TCM for Yiqi Huoxue Huatan were summarized and reviewed.

Diabetes mellitus (DM) is a chronic metabolic disease characterized by hyperglycemia. Its main complications of diabetes, such as diabetic angiopathy, have seriously affected the quality of life for patients, and have become an important cause of death and disability. The underlying pathological changes include macrovascular lesions and microvascular lesions. Diabetic macrovascular lesions mainly involve thoracic aorta, coronary artery, carotid artery, cerebral artery and peripheral blood vessels, etc., and the common clinical diseases include coronary heart disease, stroke, peripheral neuropathy, lower extremity arteriosclerosis, etc. Diabetic microvascular lesions mainly involve the heart, brain, kidney and other microvessels. Nowadays, various new oral hypoglycemic agents and insulin have emerged in the society and are widely used in clinical practice. However, traditional Chinese medicines(TCMs) have stable curative effect, less side effect, and can improve glucose metabolism, lipid metabolism, insulin resistance, oxidative stress, expression of inflammatory cytokines, vascular endothelial injury, microcirculation disorders, balance of fibrinolysis system and blood coagulation system, and improve the syndromes of TCMs, etc. They have been widely recognized and applied in the prevention and treatment of diabetic angiopathy. A profound understanding on the etiology, pathogenesis and treatment of diabetic angiopathy has been formed in Chinese medicine. Therefore, in this paper, we would summarizes the understanding on Chinese medicine for diabetic angiopathy and the mechanism of Yiqi Huoxue prescription in the treatment of diabetic angiopathy in the past three years.

Objective::To investigate the protective effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts on vascular calcification induced by high glucose in mice by observing the expression of osteopontin (OPN) and smooth muscle 22α (SM22α) as well as vascular calcium deposition in the common carotid artery and thoracic aorta of mice. Method::Totally 130 male C57BL/6 mice were randomly divided into normal control group and high glucose group. The mice in high glucose group were intraperitoneally injected with streptozotocin(STZ), and fed on a high-fat diet for 7 months. Then, the mice were randomly divided into model group, low-dose and high-dose Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts groups (0.819, 1.638 g·kg^-1), and metformin group (150 mg·kg^-1). Each group was intragastrically administered once a day for 9 weeks. The changes in blood glucose were measured. Seven days before the end of the administration, a group of 4-week old male C57BL/6 mice were purchased and fed normally for one week as a youth group. At the end of the administration, the common carotid artery and thoracic aorta tissues of the mice were collected. Von Kossa staining was used to determine the degree of calcium deposition in the common carotid artery and thoracic aorta. The expression levels of OPN and SM22α protein in the common carotid artery and thoracic aorta were detected by immunohistochemistry. The expression of OPN and SM22α protein in the common carotid artery of mice was determined by Western blot. Result::As compared with the young group, the blood glucose of the normal control group was slightly increased without statistical difference, the common carotid artery and thoracic aorta were uniformly stained, and no black granular precipitate was observed. As compared with the normal control group, the blood glucose of the model group was increased (P<0.01), with a large amount of brown-black particles deposited in the intimal elastic fibers, showing obvious calcium salt deposition. As compared with the model group, blood glucose was significantly decreased in each administration group (P<0.05, P<0.01), and the degree of vascular calcium salt deposition was significantly reduced. There were no significant changes in expression levels of OPN protein and SM22α protein in the common carotid artery and thoracic aorta between the youth group and normal control group. As compared with the normal control group, the expression of intimal OPN protein in the common carotid artery and thoracic aorta of the model group was positive, SM22α protein expression was weakly positive, and the gray value of OPN protein expression in the common carotid artery was significantly increased (P<0.01), while the gray value of SM22α protein was decreased significantly (P<0.01). As compared with the model group, the expression levels of intimal OPN protein and SM22α protein in the common carotid artery and thoracic aorta of each administration group were significantly improved, and the gray value of OPN protein expression in the common carotid artery was reduced (P<0.05, P<0.01), while SM22α protein expression was significantly increased (P<0.01). Conclusion::High glucose can induce calcification of common carotid artery and thoracic aorta in mice and accelerate vascular aging. This formation process may be related to the expression of OPN and SM22α. Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts can reduce vascular calcification and delay vascular aging by regulating the expression of OPN and SM22α.