Antiosteoporotic Activity of Saururus chinensis Extract in Ovariectomized Rats

Recent studies suggest that phytoestrogens may exert a protective effect against osteoporosis. This study examined whether treatment with phytoestrogen extracts from Saururus chinensis (SC) exerted a preventive effect on estrogen‐deficiency‐induced osteoporosis. Six‐ to seven‐month‐old female Sprague–Dawley rats were randomly assigned into either a sham‐operated group or one of three ovariectomy (OVX) subgroups: OVX treated with vehicle, OVX with alendronate, and OVX with SC extract (SC). Rats began receiving treatment 4 weeks before the OVX treatment and continued receiving treatment for an additional 10 weeks after OVX (for a combined total of 14 weeks). The results showed that the SC treatment prevented loss of femur bone mineral density after OVX, as determined by a significant decrease in the levels of serum bone turnover markers osteocalcin and alkaline phosphatase as well as urinary deoxypyridinoline. Micro‐computed tomography analysis showed that the SC treatment significantly prevented decreases in bone volume/tissue volume, trabecular number and trabecular thickness, while also preventing an increase in trabecular separation. It was concluded that SC treatment could prevent OVX‐induced loss of bone mass and deterioration in trabecular microarchitecture by suppressing bone turnover, thereby maintaining bone structural integrity. Further, no stimulation of proliferation of uterine tissue was noted. Therefore, it is suggested that treatment with S. chinensis extracts might be a potential alternative therapy for treating postmenopausal osteoporosis. Copyright © 2012 John Wiley & Sons, Ltd.     

三白草中槲皮素的薄层色谱鉴别

目的：建立三白草药材中槲皮素的提取方法,对三白草药材中槲皮素进行薄层鉴别。方法：采用硅胶G板分离三白草药材中槲皮素成分。结果：供试品色谱在与对照品色谱相应位置上显相同颜色的斑点,可用于鉴别三白草中槲皮素。结论：薄层色谱方法简便、专属性强、重现性好,可用于三白草药材的鉴别。     

Hepatoprotective flavonol glycosides of Saururus chinensis herbs

Two flavanol glycosides, quercetin-3-O -β-D -glucuronopyranoside (1) and quercetin-3-O -β-D -glucuronopyranosyl methyl ester (2), from the aerial part of Saururus chinensis which possess hepatoprotective activities have been identified. Both compounds 1 and 2 significantly reduced the level of glutamic pyruvic transaminase released from intoxicated rat hepatocytes and resulted in 78% and 82% blockade of the toxicity at 200 μM , respectively. Furthermore, these compounds also reduced the level of toxicity as measured by the release of sorbitol dehydrogenase and resulted in 76% and 83% blockade of the toxicity at 200 μM , respectively. Also, these glycosides significantly increased the activity of glutathione-S-transferase in the injured rat hepatocytes and resulted in 68% and 86% blockade of the toxicity at 200 μM , respectively. Finally, these flavonol glycosides promoted the preservation of glutathione in carbon tetrachloride-intoxicated hepatocytes by 46% and 52% of the untreated control, respectively. © 1997 John Wiley & Sons, Ltd.     

Anti‐Angiogenic Activity of Herba Epimedii on Zebrafish Embryos In Vivo and HUVECs In Vitro

Herba Epimedii, an herb commonly used in East Asian medicine, is commonly used for treatment of impotence, osteoporosis and many inflammatory conditions in traditional Chinese medicine. Recent studies revealed that Herba Epimedii also has anti‐tumor or anti‐cancer activities, which may possibly be mediated through anti‐angiogenesis. This study aims to examine and confirm the anti‐angiogenic activity in the herb using both in vivo and in vitro approaches. The 95% ethanol extract and four subsequent fractions (n‐hexane, ethyl acetate (EA), n‐butanol and aqueous fractions) of Herba Epimedii were tested on the zebrafish model by the quantitative assay for endogenous alkaline phosphatase; then, the active fraction was further tested on Tg(fli1a:EGFP)y1 zebrafish embryos and human umbilical vein endothelial cells (HUVECs) for the anti‐angiogenic effects. In addition, the action mechanism of Herba Epimedii was further investigated on wild‐type zebrafish embryos and HUVECs. The EA fraction showed anti‐angiogenic effects in both in vivo and in vitro models. Further experiments demonstrated that it might affect angiogenesis by acting on multiple molecular targets in zebrafish embryos and ERK signaling pathway in HUVECs. In conclusion, Herba Epimedii can inhibit angiogenesis, which may be the mechanism for its anti‐inflammatory, anti‐tumor and anti‐cancer actions. Copyright © 2012 John Wiley & Sons, Ltd.     

Safflower Seed Extract Inhibits Osteoclast Differentiation by Suppression of the p38 Mitogen‐activated Protein Kinase and IκB Kinase Activity

Safflower seed has been reported to have a protective effect against bone loss diseases. However, the precise molecular mechanisms underlying the inhibitory effect of safflower seed in osteoclast differentiation remain unclear. In this study, we investigated the inhibitory action of safflower seed extract (SSE) on the receptor activator of nuclear factor κB ligand (RANKL)‐induced osteoclastogenesis in cultured mouse‐derived bone marrow macrophages (BMMs). We found that SSE significantly inhibited the formation of tartrate‐resistant acid phosphatase (TRAP)‐positive multinucleated cells in BMMs without cytotoxicity. The gene expressions of nuclear factor of activated T‐cells (NFATc1) and TRAP, which are genetic markers of osteoclast differentiation, were substantially decreased by SSE in a dose‐dependent manner. Also, SSE diminished RANKL‐mediated intracellular reactive oxygen species (ROS) generation on osteoclastogenesis in a dose‐dependent manner. The SSE thereafter suppressed RANKL‐induced p38 mitogen‐activated protein kinase and IκBα kinase signalling activities which were activated by ROS generation for osteoclastogenesis. Additionally, SSE was found to decrease RANKL‐induced actin ring formation, which is required for bone resorption activity. Taken together, our results suggest that SSE acts as a RANKL‐induced osteoclastogenesis inhibitor by suppression of ROS generation. This induces a remarkable suppression of the p38 and IκBα kinase pathways, thereby suppressing the gene expression of NFATc1 in osteoclast precursors. Copyright © 2012 John Wiley & Sons, Ltd.     

猕猴桃根多糖对人胃癌SGC-7901细胞增殖、凋亡及p-p38表达的影响

目的探讨猕猴桃根多糖（Actinidia chinensis Planch polysaccharid,ACPS）对人胃腺癌细胞（SGC-7901）增殖和凋亡的影响,及对SGC-7901细胞磷酸化p38（p-p38）蛋白表达的影响。方法采用CCK-8检测不同浓度ACPS对SGC-7901细胞的24、48、72h的抑制作用;流式细胞技术检测各浓度ACPS作用48h后SGC-7901细胞凋亡的发生率;Western blot法检测各浓度ACPS作用SGC-7901细胞后前体半胱氨酰天冬氨酸酶-9（pro-caspase-9）、聚腺苷二磷酸核糖聚合酶（PARP）和p-p38蛋白量的表达,以及p38特畀性抑制剂预处理细胞后pro-caspase-9、PARP和p-p38蛋白量的表达。结果与对照组比较,1、2．5、5、10mg／mLACPS作用胃癌SGC．7901细胞后吸光度下降（P〈0．05）;同时药物剂量越高,作用时间越长,吸光度越低（P〈0．01）;24、48、72hIC50分别为7．43、3．88、1．32mg／mL;ACPS能下调SGC-7901细胞中pro-caspase-9蛋白的表达（P〈0．01）,增加PARP剪切蛋白的表达（P〈0．01）;进一步研究发现,ACPS处理SGC-7901细胞24h后,p38的磷酸化水平升高（P〈0．05）,p38特异性抑制剂处理细胞2h后能抑制p38磷酸化表达,并能抑制ACPS诱导的细胞凋亡。结论ACPS具有抑制人胃癌SGC-7901细胞增殖,诱导其凋亡的作用;激活p38途径,进而激活caspase-9和PARP,最终导致细胞死亡,可能是其诱导胃癌细胞凋亡的分子机制之一。     

In vitro Anti‐diabetic Activity and Chemical Characterization of an Apolar Fraction of Morus alba Leaf Water Extract

The tea from the white mulberry (Morus alba L.) leaf is a worldwide known traditional medicine of type II diabetes. Here, we report the investigation of the dichloromethane‐soluble fraction obtained in a 0.24% m/m yield from the hot water extract of mulberry leaves. A significant, dose‐dependent activity was found by means of the 24‐h glucose consumption of fully differentiated adipocytes both in the absence and presence of insulin. The fraction was characterized by HPLC‐DAD, GC‐MS and GC‐FID. The main constituent (40.3% by means of GC‐FID) was isolated and identified as loliolide by EIMS, HRESIMS and NMR spectroscopy. In the active fraction benzyl alcohol, ethyl benzoate, t‐cinnamic acid, p‐hydroxyacetophenone, t‐coniferyl alcohol and synapil alcohol were also identified by GC‐MS and quantified by GC‐FID (0.7, 1.3, 1.5, 2.9, 7.5 and 2.6%, respectively). Copyright © 2012 John Wiley & Sons, Ltd.     

Inhibition of UDP‐Glucuronosyltransferases (UGTs) Activity by constituents of Schisandra chinensis

Structure–activity relationship for the inhibition of Schisandra chinensis's ingredients toward (Uridine‐Diphosphate) UDP‐glucuronosyltransferases (UGTs) activity was performed in the present study. In vitro incubation system was employed to screen the inhibition capability of S. chinensis's ingredients, and in silico molecular docking method was carried out to explain possible mechanisms. At 100 μM of compounds, the activity of UGTs was inhibited by less than 90% by schisandrol A, schisandrol B, schisandrin, schisandrin C, schisantherin A, gomisin D, and gomisin G. Schisandrin A exerted strong inhibition toward UGT1A1 and UGT1A3, with the residual activity to be 7.9% and 0% of control activity. Schisanhenol exhibited strong inhibition toward UGT2B7, with the residual activity to be 7.9% of control activity. Gomisin J of 100 μM inhibited 91.8% and 93.1% of activity of UGT1A1 and UGT1A9, respectively. Molecular docking prediction indicated different hydrogen bonds interaction resulted in the different inhibition potential induced by subtle structure alteration among schisandrin A, schisandrin, and schisandrin C toward UGT1A1 and UGT1A3: schisandrin A > schisandrin > schisandrin C. The detailed inhibition kinetic evaluation showed the strong inhibition of gomisin J toward UGT1A9 with the inhibition kinetic parameter (K_i) to be 0.7 μM. Based on the concentrations of gomisin J in the plasma of the rats given with S. chinensis, high herb–drug interaction existed between S. chinensis and drugs mainly undergoing UGT1A9‐mediated metabolism. In conclusion, in silico‐in vitro method was used to give the inhibition information and possible inhibition mechanism for S. chinensis's components toward UGTs, which guide the clinical application of S. chinensis. Copyright © 2015 John Wiley & Sons, Ltd.     

独一味注射液抗炎作用及其机制

目的：独一味注射液的抗炎作用的研究及其作用机制的初步探讨。方法：通过大鼠棉球肉芽肿模型和二甲苯所致小鼠耳肿胀模型,观察独一味注射液的抗炎作用;采用中性红吞噬试验研究独一味注射液对小鼠巨噬细胞的吞噬功能的影响,并采用小鼠胸腺细胞增殖法检测独一味注射液对小鼠巨噬细胞上清液中的白介素-1含量的影响。结果：独一味注射液在0．225和0．45g．kg^-1剂量下能显著的抑制大鼠棉球肉芽肿的形成,在0．45,0．9和1．8g．kg。剂量下能显著的抑制二甲苯所致的小鼠耳肿胀;独一味注射液显著的增强了巨噬细胞的吞噬能力,并且抑制了脂多糖诱导的白介素-1的分泌。结论：独一味注射液具有明显的抗炎作用,其作用机制可能与增强巨噬细胞的吞噬功能和抑制白介素-1的分泌有关。     

In vitro and in vivo antiproliferative activity of laherradurin and cherimolin‐2 of annona diversifolia saff

Acetogenins from Annonaceae (ACG) are potent inhibitors of the mitochondrial complex I, they present cytotoxic activity on neoplasic lines, including those with multiresistance to drugs. In vivo antitumor activities of some ACG have been reported; however, no information is available regarding the relationship between their cytotoxic activity in cell cultures and their antiproliferative action in vivo. Laherradurin and cherimolin‐2 acetogenins were isolated from Annona diversifolia seeds, and their inhibitory potential was analysed in vitro on HeLa and SW‐480 cells. Doses containing 1, 10, 100 and 500 times the IC₅₀ obtained from the proliferation assays and multiplied by the weight of the animal, were injected once daily into athymic mice bearing these cancer cell lines; their effect upon tumor growth was measured over a period of 20 days. Laherradurin was more active than cherimolin‐2, and it showed in in vitro proliferation assays a similar IC₅₀ in both neoplasic lines. In athymic mice, laherradurin administered in 10×, 100× and 500× doses, reduced the size of HeLa tumors, and with 100× and 500× doses, affected the SW‐480 tumor development. These doses were similar to results found with the control drug doxorubicin (p ≤ 0.05). On the other hand, cherimolin‐2 had an effect on HeLa tumors cells at 100× and 500× doses (p ≤ 0.05). Copyright © 2009 John Wiley & Sons, Ltd.     

蒲葵籽提取物体外抗HIV-1活性及机制的初步研究

目的研究蒲葵籽提取物的体外抗HIV-1活性,对有活性粗提物进行初步机制研究。方法采用细胞毒性、合胞体抑制、HIV-1感染细胞保护实验和HIV-1 p24抗原测定等实验对蒲葵籽提取物体外抗HIV-1活性进行筛选和确认;采用重组HIV-1逆转录酶和蛋白酶活性抑制实验,融合阻断实验初步探讨活性粗提物的作用机制。结果蒲葵籽的醋酸乙酯(P3)提取物具有较强的体外抗HIV-1活性,P3抑制HIV-1诱导C8166细胞形成合胞体的EC50为5.64μg/mL,对C8166细胞的毒性较小,CC50大于200μg/mL,治疗指数(TI)大于35.46;P3抑制HIV-1急性感染中p24抗原表达的EC50为23.04μg/mL,抑制正常C8166细胞与慢性感染细胞H9/HIV-1 B融合的EC50为8.00μg/mL;P3在质量浓度为200μg/mL时,对HIV-1逆转录酶的抑制率为28.86%;P3抑制重组HIV-1蛋白酶活性的EC50为1.77μg/mL。结论蒲葵籽的醋酸乙酯提取物(P3)具有较强的体外抗HIV-1活性,其作用机制可能主要为阻断病毒进入和抑制HIV-1蛋白酶活性。     

针毛蕨的化学成分及其体内外抗肿瘤活性研究

 目的 研究针毛蕨（Macrothelypteris oligophlebia化学成分及其体内外抗肿瘤活性。方法 采用各种色谱技术进行分离纯化,根据理化性质和波谱数据进行结构鉴定,以MTT法测试各化合物对肿瘤细胞的体外抑制作用,并观察针毛蕨提取物对S₁₈₀肉瘤小鼠肿瘤生长的影响。结果 分离得到7个黄酮类化合物,分别鉴定为：荚果蕨素（1、柚皮素（2、柚皮素-5-O-β-D-吡喃葡萄糖苷（3、原芹菜素（4、去甲氧基荚果蕨素（5、芹菜素（6和原芹菜素-4′-O-β-D-葡萄糖苷（7。结论 所有化合物均首次从针毛蕨中分离得到;化合物4对肿瘤细胞增殖有显著的抑制作用,且呈现出良好的剂量依赖性;体内抗肿瘤试验显示针毛蕨提取物对S₁₈₀肉瘤小鼠肿瘤的生长有明显的抑制作用。     

不同寄主植物对菟丝子、南方菟丝子多糖含量的影响

目的:研究不同寄主上的菟丝子、南方菟丝子中多糖的含量。方法:用苯酚-浓硫酸法测定含量。结果:寄生于大豆上的菟丝子及南方菟丝子多糖含量较高。菟丝子及南方菟丝子种间多糖的含量差异不明显。随着菟丝子种子成熟程度的增加,多糖及可溶性糖的含量均增加。结论:寄主植物影响菟丝子及南方菟丝子中多糖的含量。     

珍珠梅提取物的体内抑瘤作用及其机理研究

目的：探讨珍珠梅醋酸乙酯提取物对S180荷瘤小鼠体内的肿瘤抑制作用及机理。方法：按照标准方法给36只小鼠接种S180肉瘤后，给予珍珠梅提取物10 d，观察瘤重及体重，免疫组织化学方法检测肿瘤组织中的hcl-2和p53蛋白，比色分析法检测血清SOD，GSH-Px的活性及MDA含量的变化。结果：给予珍珠梅醋酸乙酯提取物的小鼠瘤重明显小于对照组，实验组与对照组体重变化无显著性差异：对照组的bcl-2和p53蛋白的表达率明显高于实验组，实验组的SOD，GSH-Px活性高于对照组，而MDA含量低于对照组。结论：珍珠梅醋酸乙酯提取物对小鼠S180肿瘤有抑制作用，并有可能影响bcl-2和P53的表达及清除体内自由基。     

Transcriptomic Analysis Reveals Wound Healing of Morus alba Root Extract by Up‐Regulating Keratin Filament and CXCL12/CXCR4 Signaling

Facilitation of the wound healing process is important because a prolonged wound site increases pain and the risk of infection. In oriental medicine, an extract of Morus alba root (MA) has usually been prescribed as traditional treatment for accelerating wound healing, and it has been proven to be safe for centuries. To study the molecular mechanism of MA‐mediated skin wound healing, we performed a primary cell culture and a skin explant culture and observed significant difference between the groups with and without MA extract. In the cellular system, a real‐time cell analysis and real‐time quantitative PCR were performed. It was found that MA extract enhanced proliferation in a dose‐dependent manner on Kera‐308 cell line, and up‐regulated keratin expression including wound‐induced Krt6a. In skin explant culture, the mRNA level derived from cell outgrowth displayed a tendency toward more up‐regulated mRNA associated keratin filaments and toward a more up‐regulated mRNA level of C‐X‐C motif chemokine 12 (CXCL12) and a chemokine receptor 4 (CXCR4) axis signaling pathway downstream. In this process, we concluded that MA extract had a scientific possibility of wound repair by increasing intracellular and extracellular supports and by inducing a CXCL12/CXCR4 signaling pathway. Copyright © 2015 John Wiley & Sons, Ltd.     

ESP‐102, a combined extract of Angelica gigas,Saururus chinensis and Schizandra chinensis,protects against glutamate‐induced toxicity in primary cultures of rat cortical cells

It was reported previously that ESP‐102, a combined extract of Angelica gigas, Saururus chinensis and Schizandra chinensis, significantly improved scopolamine‐induced memory impairment in mice and protected primary cultured rat cortical cells against glutamate‐induced toxicity. To corroborate this effect, the action patterns of ESP‐102 were elucidated using the same in vitro system. ESP‐102 decreased the cellular calcium concentration increased by glutamate, and inhibited the subsequent overproduction of cellular nitric oxide and reactive oxygen species to the level of control cells. It also preserved cellular activities of antioxidative enzymes such as superoxide dismutase, glutathione peroxidase and glutathione reductase reduced in the glutamate‐injured neuronal cells. While a loss of mitochondrial membrane potential was observed in glutamate treated cells, the mitochondrial membrane potential was maintained by ESP‐102. These results support that the actual mechanism of neuroprotective activity of ESP‐102 against glutamate‐induced oxidative stress might be its antioxidative activity. Copyright © 2009 John Wiley & Sons, Ltd.     

Curcumin I Mediates Neuroprotective Effect Through Attenuation of Quinoprotein Formation,p‐p38 MAPK Expression,and Caspase‐3 Activation in 6‐Hydroxydopamine Treated SH‐SY5Y Cells

6‐Hydroxydopamine (6‐OHDA) selectively enters dopaminergic neurons and undergoes auto‐oxidation resulting in the generation of reactive oxygen species and dopamine quinones, subsequently leading to apoptosis. This mechanism mimics the pathogenesis of Parkinson's disease and has been used to induce experimental Parkinsonism in both in vitro and in vivo systems. In this study, we investigated the effects of curcumin I (diferuloylmethane) purified from Curcuma longa on quinoprotein production, phosphorylation of p38 MAPK (p‐p38), and caspase‐3 activation in 6‐OHDA‐treated SH‐SY5Y dopaminergic cells. Pretreatment of SH‐SY5Y with curcumin I at concentrations of 1, 5, 10, and 20 μM, significantly decreased the formation of quinoprotein and reduced the levels of p‐p38 and cleaved caspase‐3 in a dose‐dependent manner. Moreover, the levels of the dopaminergic neuron marker, phospho‐tyrosine hydroxylase (p‐TH), were also dose‐dependently increased upon treatment with curcumin I. Our results clearly demonstrated that curcumin I protects neurons against oxidative damage, as shown by attenuation of p‐p38 expression, caspase‐3‐activation, and toxic quinoprotein formation, together with the restoration of p‐TH levels. This study provides evidence for the therapeutic potential of curcumin I in the chemoprevention of oxidative stress‐related neurodegeneration. Copyright © 2013 John Wiley & Sons, Ltd.     

余甘子提取物体外抗单纯疱疹病毒1型和2型的初步研究

目的 从余甘子的醋酸乙酯提取物、甲醇提取物和水提物3种提取物体外筛选抗单纯疱疹病毒1型和2型的活性成分.方法 通过观察药物毒性、病毒引起的细胞病变效应、空斑减数法,判断药物的毒性及其抗病毒效应.结果 余甘子的醋酸乙酯提取物、甲醇提取物和水提物对于单纯疱疹病毒1型的半数抑制浓度分别为25.28,66.17,100.94 μg/ml;余甘子的3种提取物对于单纯疱疹病毒2型的半数抑制浓度分别为31.70,180.3,112.1 μg/ml.结论 余甘子3种提取物在体外对单纯疱疹病毒1型和2型均有一定的抑制作用.