Effect of Ginseng Radix et Rhizoma， Notoginseng Radix et Rhizoma， and Chuanxiong Rhizoma Extract on Mitochondrial Oxidative Stress in Hydrogen Peroxide-induced Endothelial Cell Aging / 中国实验方剂学杂志

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-<italic>β</italic>-galactosidase （SA-<italic>β</italic>-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-<italic>β</italic>-gal staining results showed that H₂O₂ group had increased blue-stained cells compared with other groups （<italic>P</italic><0.01）. Compared with those in the control group， the ratio of G₀/G₁ phase cells significantly increased （<italic>P</italic><0.05） and that of G₂/M phase cells decreased （<italic>P</italic><0.05） in the H₂O₂ group. Compared with those in the H₂O₂ group， the proportion of G₀/G₁ cells decreased （<italic>P</italic><0.05） while that of G₂/M cells increased （<italic>P</italic><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 （<italic>P</italic><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 （<italic>P</italic><0.01）， as well as the Resv group （<italic>P</italic><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 （<italic>P</italic><0.05）， whereas the protein level of p-p66 showed an opposite trend （<italic>P</italic><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.

Mechanism of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma on SIRT1 autophagy pathway of endothelial cell senescence induced by hydrogen peroxide / 中国中药杂志

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.

Research Progress of Traditional Chinese Medicine for Yiqi Huoxue Huatan in Treatment of Atherosclerosis / 中国实验方剂学杂志

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.

Investigation of clinical practice and side effects of Shenfu injection / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the status of clinical practices and side effects related to Shenfu injection.</p><p><b>METHOD</b>It was searched the Chinese medical citation index (CMCI) database and China national knowledge infrastructure (CNKI) database using the word "Shenfu injection". Original clinical studies of Shenfu injection which were published from January 1 987 to March 2012 in Chinese magazines were included.</p><p><b>RESULT</b>A total of 1 433 relevant clinical studies were identified. It was investigated and analyzed the side effects of Shenfu injection. For cases with side effects, the diseases for treatment, the medication dosage, the course, and menstrum of treatment for the side effects were evaluated. All the side effects were mild except five cases of serious adverse reaction.</p><p><b>CONCLUSION</b>Shenfu injection has been widely used in clinical patients since it came into the market in 1987. However, there are some problems in its clinical practices, such as deficiency in selecting and expanding diseases for treatment, evaluating the dosage and treatment method. Therefore, it is imperative to evaluate drugs by literature and good clinical application after the drugs come into the market with evidence-based medicine.</p>