Ethanol extract of Fructus Ligustri Lucidi promotes osteogenesis of mesenchymal stem cells

Fructus Ligustri Lucidi (FLL) has been used in traditional Chinese medicine for over 1000 years. The ethanol extract of FLL (EFLL) has been shown to be a potential candidate in the prevention and treatment of osteoporosis. The present study aimed to determine whether EFLL carries out the effect by promoting osteogenesis in mesenchymal stem cells (MSCs). The osteogenic differentiation of MSCs was evaluated by their alkaline phosphatase (ALP) activities and mineralization. Expression of genes was detected by RT‐PCR. We found that EFLL significantly stimulated the ALP activities and shortened the time needed for the mineralization of MSCs during osteogenic differentiation. The expression of several osteoblast differentiation regulators was also upregulated by EFLL during this process. Our study demonstrated that the EFLL is capable of enhancing osteogenic differentiation of MSCs. It might be useful for treating diseases with inadequate bone formation, including osteoporosis. Copyright © 2009 John Wiley & Sons, Ltd.     

Inhibition of Osteoclast Differentiation by Ginsenoside Rg3 in RAW264.7 Cells via RANKL,JNK and p38 MAPK Pathways Through a Modulation of Cathepsin K: An In Silico and In Vitro Study

Various studies have demonstrated that overexpression of cathepsin K (Cat‐K) causes excessive bone loss, which ultimately leads to a variety of bone diseases including osteoporosis. Therefore, inhibition of Cat‐K signifies a potential therapeutic target in osteoporosis treatment. Ginsenoside Rg3 is one of the most promising compound of Panax ginseng Meyer (P. ginseng) with numerous biological activities. Thus, in recent study the inhibitory effect of Rg3 isolated from P. ginseng was investigated in order to impede the osteoclast activity by an in silico approach followed by in vitro study validation using RAW264.7 cells through the investigation of different biological activity prediction such as absorption distribution metabolism and excretion (ADMET) properties against Cat‐K protein. The docking results of our study showed that Rg3 is a non‐toxic compound and may act as a drug‐like molecule. Additionally, the molecular interaction of Rg3 with the active residues of Cat‐K markedly describes its inhibitory effects on osteoclastogenesis. Findings of the present study exhibited that Rg3 significantly reduced receptor activator of nuclear factor kappa B ligand (RANKL)‐induced tartrate‐resistant acid phosphatase (TRAP) activity, pit formation (actin rings), and TRAP‐positive multinucleated cells development in RAW264.7 cells. Furthermore, Rg3 dose‐dependently reduced the mRNA expression levels of osteoclast‐specific markers such as RANK, TRAP, and Cat‐K induced by RANKL through the down regulation of p38, extracellular signal‐regulated kinase, and c‐Jun N‐terminal kinase (JNK) pathways. In conclusion, in silico docking study and in vitro validation together suggested that Rg3 inhibits osteoclastogenesis and reduces bone resorption through the inhibition of Cat‐K. Therefore, Rg3 might be a useful therapeutic agent for the treatment of osteoporosis and proper bone formation. Copyright © 2015 John Wiley & Sons, Ltd.     

Inhibitory effect of sulphated polysaccharide porphyran (isolated from Porphyra yezoensis) on RANKL‐induced differentiation of RAW264.7 cells into osteoclasts

Safe and efficient therapeutic agents for bone diseases are required in natural sources. We previously found that edible seaweed‐derived polysaccharide porphyran exhibited anti‐inflammatory effects through the down regulation of nuclear factor‐κB. The aim of this study was to investigate the availability of porphyran as a therapeutic agent for bone diseases. The effects of porphyran on receptor activator of nuclear factor κB ligand (RANKL)‐induced osteoclastogenesis in RAW264.7 cells were examined. Porphyran suppressed RANKL‐induced osteoclast formation in a concentration‐dependent manner (6.25–50 μg/ml) without any cytotoxic effects. Furthermore, real‐time polymerase chain reaction analyses indicated that porphyran at 50 μg/ml significantly attenuated the RANKL‐induced increase in the mRNA levels of osteoclastogenesis‐related marker genes such as nuclear factor of activated T cells, tartrate‐resistant acid phosphatase, cathepsin K, and matrix metalloproteinase‐9 in RAW264.7 cells. To our knowledge, this is the first report showing that edible‐seaweed‐derived polysaccharide porphyran can suppress RANKL‐induced osteoclastogenesis. Our results suggest that porphyran can be used as a safe therapeutic agent to improve osteoclast‐related pathological conditions.     

淫羊藿-女贞子对GIOP大鼠骨密度和激素水平的相关性分析

目的: 研究糖皮质激素性骨质疏松（GIOP）大鼠内源性激素水平与骨密度的相关性,探讨淫羊藿-女贞子配伍抗GIOP的作用机制。 方法: SD大鼠60只,雌雄各半,随机分为正常对照组、模型对照组、女贞子组、淫羊藿组、淫羊藿-女贞子组、钙尔奇组。地塞米松1 mg·kg^-1,im,2次／周,建立骨质疏松模型。造模同时各给药组灌服相应药物,女贞子组灌服女贞子水煎液9.5 g·kg^-1,淫羊藿组灌服淫羊藿水煎液9.5 g·kg^-1,淫羊藿-女贞子组灌服淫羊藿-女贞子（4∶3）水煎液9.5 g·kg^-1,钙尔奇组灌服钙尔奇碳酸钙D3片0.277 3 g·kg^-1。8周后处死取材,放射免疫法检测血清皮质醇（COR）、睾酮（T）、雌二醇（E₂）含量;双能X射线骨密度仪测量离体股骨骨密度（BMD）;并对实验数据进行相关性分析。 结果: 模型对照组大鼠血清COR,T,E₂含量以及股骨不同部位BMD均显著降低（P<0.05或0.01）;与模型对照组比较,各给药组均可显著上调血清COR水平（P<0.05或0.01）,淫羊藿-女贞子组血清T含量显著升高（P<0.05）,女贞子组、淫羊藿-女贞子组血清E₂含量显著升高（P<0.05或0.01）,且各用药组不同部位BMD均较模型对照组显著升高（P<0.05或0.01）;相关性分析结果显示BMD与血清COR,T,E₂水平呈显著正相关（P<0.05或0.01）。 结论: GIOP大鼠BMD与内源性激素水平密切相关,淫羊藿-女贞子配伍对激素水平的影响可能是其防治GIOP的机制之一。     

Systemic effect of Fructus Psoraleae extract on bone in mice

Fructus Psoraleae extract is used in China for the treatment of bone diseases. The objective of the study was to investigate the systemic effect of Fructus Psoraleae extract consumption on bone histomorphology. Sixteen 8‐week‐old male BALB/c mice were divided into control and experimental groups. In the control group, eight mice were fed daily with distilled water. In the experimental group, eight mice were fed daily with distilled water mixed with Fructus Psoraleae extract. The mice were kept for 5 weeks and then killed. Using micro‐computed tomography, 20 micro‐tomographic slices with a separation of 0.25 mm were acquired to cover the proximal end of the left tibia of each mouse. Quantitative morphometry of the bone structure was performed. The results showed that consumption of Fructus Psoraleae extract significantly increased the bone volume/tissue volume ratio by 11.8%. The bone trabeculae increased by 7.1% in thickness so that the bone density was increased. To conclude, Fructus Psoraleae extract taken orally increases bone density and alters bone histomorphology. Copyright © 2010 John Wiley & Sons, Ltd.     

南北五味子化学成分、药理作用等方面差异的研究进展

五味子为木兰科植物五味子Schisandra chinesis或华中五味子S.sphenanthera的干燥成熟果实,前者称为"北五味子",后者称为"南五味子"。五味子始载于《神农本草经》,作为中药使用已有一千多年历史,且应用于多种成方制剂。早在明代南北五味子的疗效差异已被众多医家认识,自2000年版《中国药典》开始,对五味子和南五味子分别收载,并建立了各自的质量控制标准,直至2015年版《中国药典》对其不断完善,南、北五味子形态、产地不同,成分各有差异,功能主治各有侧重,但目前临床使用存在南北五味子混用之现象。所以为了区分南北五味子,更加合理、安全、有效地将五味子应用于现代临床的治疗,该文对近年来关于南北五味子的研究进行搜集、检索、归纳、总结,从品种鉴别,化学成分(其中北五味子挥发油成分、总木脂素的含量、多糖疗效均大于南五味子),药理作用(以五味子对中枢神经系统、消化系统、心血管系统的影响为主)等方面的差异进行综述,以期为南、北五味子的研究开发以及临床应用提供参考,使南北五味子不但从形式上而且从根本上真正分开,使临床用药更加安全、准确。     

The effects of hypoglycemia and weight loss of total lignans from Fructus Arctii in KKAy mice and its mechanisms of the activity

Fructus Arctii (great burdock achene) is the dried ripe fruit of Arctium lappa L. (family Asteraceae) and is included in the Chinese pharmacopoeia. It has been reported that the clinical use of Fructus Arctii resulted in a satisfactory hypoglycemic effect in diabetic patients. This study aimed to investigate antidiabetic activity and mechanism of total lignans from Fructus Arctii (TLFA) in KKAy mice, a spontaneous Type 2 diabetic rodent model that exhibits marked obesity. In this study, KKAy mice were gavaged once daily with solvents (0.5% sodium carboxymethyl cellulose), TLFA (250 and 125 mg/kg), or metformin (200 mg/kg) for 11 weeks, and C57BL/6J mice treated with saline solution (0.9%, w/v) were used as normal control. The results indicate that TLFA has dual effects of hypoglycemia and weight loss, and administration of TLFA in KKAy mice could decrease fasting blood glucose, glycated hemoglobin, and body weight; improve oral glucose tolerance; increase high density lipoprotein cholesterol; and decrease triglycerides and free fatty acid in mice serum. Its efficacy may associate with multiple mechanisms of action such as stimulation of insulin secretion, activation of phosphatidylinositol 3‐kinase/protein kinase B, and adenosine‐monophosphate‐activated protein kinase signaling pathway, decreasing leptin.     

Antiinflammatory effects of Epimedium brevicornum water extract on lipopolysaccharide‐activated RAW264.7 macrophages

Epimedium brevicornum Maxim (Berberidaceae) possesses estrogenic properties. It is one of the most widespread herbal remedies used in Oriental medicine. The present study investigated the effects of Epimedium brevicornum water extract (EB) on proinflammatory mediators secreted from lipopolysaccharide (LPS)‐induced RAW264.7 macrophages. EB significantly inhibited the production of nitric oxide (NO), interleukin (IL)‐3, IL‐10, IL‐12p40, interferon‐inducible protein‐10, keratinocyte‐derived chemokine, vascular endothelial growth factor, monocyte chemotactic protein‐1 and granulocyte macrophage‐colony stimulating factor in LPS‐induced RAW264.7 cells at concentrations of 25, 50, 100 and 200 μg/mL (p < 0.05). These results suggest that EB has antiinflammatory activity related to its inhibition of NO, cytokine, chemokine and growth factor production in macrophages. Copyright © 2010 John Wiley & Sons, Ltd.     

五味子与南五味子药材的DNA条形码鉴定

目的:利用内转录间隔区2(ITS2)序列对五味子及南五味子药材进行DNA条形码鉴定。方法:对五味子和南五味子样品的ITS2序列进行聚合酶链式反应(PCR)扩增和测序。从Gen Bank中下载了五味子属10个近缘种和2个外类群的ITS2序列。利用Codon Code Aligner软件拼接,采用MEGA 5.10计算相关数据,基于K2P模型构建聚类树(NJ树),应用Blast法进行鉴定分析。获取所测样品序列的ITS2二级结构信息,分析各样品间ITS2序列二级结构的差异。结果:五味子、南五味子样品序列长度分别为231,225~227 bp,Blast鉴定五味子药材原植物为五味子Schisandra chinensis,南五味子药材原植物为华中五味子S.sphenanthera。五味子与南五味子药材的种间K2P遗传距离(0.010 4~0.015 7)远大于五味子种内K2P遗传距离(0~0.002 5)和南五味子种内K2P遗传距离(0~0.005 1)。在NJ树模型中,五味子样品被独聚为1支,南五味子样品被独聚为1支,皆表现出单系性,Bootstrap支持率较高,可与五味子属近缘种区分开。结论:以ITS2序列为标准的DNA条形码能够有效地鉴定五味子和南五味子药材。     

枳实化学成分与治疗功能性消化不良关联性分析及实验验证

目的 研究枳实治疗功能性消化不良（functional dyspepsia,FD）的药效物质基础。方法 运用超高效液相色谱-四极杆飞行时间质谱联用（UPLC-Q-TOF-MS/MS）技术对10批枳实水煎液进行分析,确定共有成分;采用不规则喂养和夹尾刺激建立FD大鼠模型,以小肠碳末推进率、胃排空率及血清中胃动素、胃泌素水平为药效指标,采用灰色关联度分析法和Pearson双变量相关性分析建立色谱图中共有峰峰面积与药效指标的相关性,筛选出枳实治疗FD的药效物质,并通过斑马鱼肠蠕动模型进行药效验证。结果 10批枳实水煎液中共指认35个共有成分,通过灰色关联度和Pearson双变量相关性分析,筛选出辛弗林、马尔敏、黄柏酮、N-甲基酪胺、圣草次苷、柚皮苷、橙皮苷和新橙皮苷共8个成分为枳实治疗FD的潜在药效物质。选择药效相关性较强、易得、可测的圣草次苷、柚皮苷、橙皮苷、新橙皮苷和辛弗林进行实验验证,发现这5个成分均显著促进斑马鱼肠道蠕动（P＜0.01）。结论 圣草次苷、柚皮苷、橙皮苷、新橙皮苷和辛弗林为枳实治疗FD的药效物质。     

砂仁及其伪品鉴别方法研究进展

砂仁是常用中药，也是我国“四大南药”之一，市场需求量巨大，而在云南、广东等地的种植量不能满足市场需求，仍需大量进口。由于进口国家豆蔻属植物种类丰富，进口砂仁中掺杂大量近缘种，药材鉴别困难，导致如今砂仁药材市场掺伪现象严重。通过查阅了近40年的文献，对目前市场上主要砂仁伪品进行总结，分析了鉴别砂仁真伪品的主要方法。随着砂仁伪品不断变化，传统性状鉴别、显微鉴别、色谱方法等很难实现砂仁的真伪鉴别，分子鉴别方法能够基本实现砂仁真伪鉴别，市场推广应用不足。若要规范砂仁药材市场，需在药材源头控制的基础上，开发更为简易、普适的鉴别方法，同时也为砂仁临床用药安全提供保障。     

Effects of Fructus schizandrae on cycloheximide‐induced amnesia in rats

The effects of Fructus Schisandrae (Schizandra Chinensis, (FS) on cycloheximide (CXM)‐induced am‐­nesia by using a passive avoidance task were studied in rats. FS at 0.25 and 0.75 g/kg administered for 1 week significantly prolonged the CXM‐shortened step‐through latency (STL). Of the fractions (n‐hexane, chloroform and water), only the water fraction at 25 mg/kg administered for 1 week prolonged the CXM‐shortened STL. These results suggest that the water fraction is the main active fraction of FS. Copyright © 1999 John Wiley & Sons, Ltd.     

小茴香、广木香抗川楝子急性肝毒性的实验研究

目的探讨小茴香、广木香对川楝子所致的急性肝损伤大鼠的保护作用及其机制。方法将SD大鼠随机分为正常组、川楝子组、川茴组、川广组,各组分别单次灌服蒸馏水、川楝子、川楝子+小茴香、川楝子+广木香的水提醇沉液。给药2 h后检测血清ALT、AST活性,计算肝脏指数,进行肝组织病理学检查,测定肝组织SOD、GSH-Px活性及MDA含量。结果与正常组比较,川楝子组ALT、AST、肝脏指数及MDA均明显升高,SOD/MDA及GSH-Px酶活性明显下降,差异有统计学意义(P<0.05,P<0.01)。与川楝子组比较,川茴组ALT、AST明显降低,SOD、SOD/MDA明显升高,差异有统计学意义(P<0.05,P<0.01);川广组MDA降低,SOD/MDA升高,差异有统计学意义(P<0.01)。川茴组与川广组对肝脏指数无明显影响。各组组织病理学改变均不明显。结论小茴香、广木香可减轻川楝子引起急性肝损伤,尤以小茴香作用为强,其作用机制可能与清除自由基、抑制脂质过氧化有关。     

Schisandra chinensis extract ameliorates age‐related muscle wasting and bone loss in ovariectomized rats

Exercise and healthy diet consumption support healthy aging. Schisandra chinensis (Turcz.) also known as “Baill.” has anti‐inflammatory and antioxidant properties. However, the role of S. chinensis as an antiaging compound has yet to be demonstrated. This study elucidated the antiaging effect of S. chinensis ethanol–hexane extract (C1) and the effect of C1 treatment on muscle and bone following physical exercise in ovariectomized (OVX) rats. RAW 264.7, human diploid fibroblasts (HDFs), C2C12 myoblasts, bone marrow macrophages, and MC3T3‐E1 cells were used for in vitro, and muscle and bone of OVX rats were used for in vivo study to demonstrate the effect of C1. The C1 significantly inhibited the expression of inflammatory molecules, β‐galactosidase activity, and improved antioxidant activity via down‐regulation of reactive oxygen species in RAW 264.7 and aged HDF cells. The C1 with exercise improved muscle regeneration in skeletal muscle of OVX rats by promoting mitochondrial biogenesis and autophagy. C1 induced osteoblast differentiation, and C1 + exercise modulated the bone formation and bone resorption in OVX rats. C1 exhibited anti‐inflammatory, antioxidant, myogenic, and osteogenic effects. C1 with exercise improved age‐related muscle wasting and bone loss. Therefore, S. chinensis may be a potential prevent agent for age‐related diseases such as sarcopenia and osteoporosis.     

牛蒡子对运动训练大鼠睾酮及相关激素含量和抗运动疲劳能力的影响

目的:研究牛蒡子对运动训练大鼠睾酮及相关激素含量和抗疲劳能力的影响。方法:以大强度耐力训练大鼠为模型,55只42 d龄雄性Wistar大鼠为对象,随机分为5组:静止组(C组)、运动组(M组)、运动+低剂量牛蒡子组(AML组),运动+中剂量牛蒡子组(AMM组),运动+高剂量牛蒡子组(AMH组),每组10只(剔除不符合实验要求的大鼠5只)。每天灌胃ig给药1次,牛蒡子组剂量分为0.5,1,3 g·kg-1,ig体积为5 m L·kg-1,C,M组ig等量生理盐水。42 d力竭游泳训练后,测定体重、力竭游泳时间及血睾酮等生化指标。结果:体重,运动组小于静止组(P<0.05);牛蒡子各组>运动组(P<0.05)。力竭游泳时间,运动组与静止组无明显差异;牛蒡子各组明显长于运动组(P<0.01),且随剂量增大而延长。血清睾酮水平,运动组为(3.51±1.46)nmol·L-1低于静止组(P<0.01);牛蒡子低、中、高剂量组分别为(4.85±1.54),(4.97±1.47),(5.07±1.56)nmol·L-1,高于运动组(P<0.01)。血清皮质酮水平,各组间均无显著差异。血清睾酮与皮质酮比值变化与睾酮变化较为一致。血清促黄体生成素、促卵泡激素水平,静止组与运动组无显著差异;牛蒡子各组高于运动组(P<0.05)。结论:补充牛蒡子可以从多靶点、多途径纠正由于运动导致的下丘脑-垂体-性腺轴功能的紊乱,有效预防血清睾酮水平的降低,进而增强抗疲劳能力。     

Synergistic anti‐inflammatory effects of quercetin and catechin via inhibiting activation of TLR4–MyD88‐mediated NF‐κB and MAPK signaling pathways

The synergistic anti‐inflammatory effect of quercetin and catechin was investigated using lipopolysaccharide (LPS)‐stimulated macrophage RAW 264.7 cells. Results showed that the combined treatment of quercetin with catechin synergistically attenuated LPS‐stimulated increase of some proinflammatory molecules, including nitric oxide, tumor necrosis factor α, interleukin‐1β, nitric oxide synthase, and cyclooxygenase‐2. Moreover, it exhibited significantly (p < 0.05) stronger inhibitory effect on nuclear translocation of nuclear factor‐κB (NF‐κB) by suppressing the phosphorylation of NF‐κB p65 and p50 submits and on the phosphorylation of ETS domain‐containing protein and c‐Jun N‐terminal kinase than any of quercetin or catechin alone. Besides, the cotreatment of quercetin with catechin significantly (p < 0.05) restored the impaired expression of toll‐like receptor 4, myeloid differentiation primary response gene 88, and some downstream effectors (IRAK1, TRAF6, and TAK1). These results suggest that quercetin and catechin possessed synergistic anti‐inflammatory effects, which may be attributed to their roles in suppressing the activation of TLR4–MyD88‐mediated NF‐κB and mitogen‐activated protein kinases signaling pathways.     

中药吴茱萸RP-HPLC指纹图谱的研究

 目的建立吴茱萸饮片的RP-HPLC指纹图谱研究方法。方法Alltech C18色谱柱;梯度洗脱;流速:1.0 mL·min^-1;检测波长:254 nm;柱温:40℃。结果该方法具有良好的稳定性和重复性,19个产地吴茱萸饮片及其炮制品共标定了15个共有峰。结论利用吴茱萸RP-HPLC指纹图谱分析法可较全面的反映吴茱萸饮片的内在质量,可用于吴茱萸饮片的质量控制。     

补骨脂药材的质量变化规律和标准探讨

目前,《中华人民共和国药典》 2020年版只选取补骨脂素和异补骨脂素作为含量测定指标成分来控制补骨脂药材质量。但是,补骨脂苷与苷元存在内部转化的问题,《中华人民共和国药典》 2020年版中补骨脂质量评价方法和标准并不能完全反映补骨脂药材的质量。通过对补骨脂的市场调研情况,结合前期试验整合分析,探讨了补骨脂的质量标准现状及质量问题。并基于药材指标性成分动态变化,提出了更加合理的质量控制方法,为补骨脂含量测定标准制订提供研究思路和依据。     

Isolation of Anti‐Inflammatory Fractions and Compounds from the Root of Astragalus membranaceus

Foot ulceration, if not treated properly, will eventually result in amputation. Inflammation may impede the wound healing process if not properly controlled. The root of Astragalus membranaceus (AR) is one of the Chinese herbs commonly found in Chinese herbal formulae used for treating foot ulcer. In this study, we aimed to identify the active fractions and/or compounds from AR aqueous extract, which are responsible for the anti‐inflammatory effect using in vitro bioassay‐guided fractionation. The anti‐inflammatory effect was monitored by the inhibition of nitric oxide (NO) released from lipopolysaccharide‐stimulated mouse macrophage RAW 264.7 cells after treated with AR aqueous extract or its fractions and isolated components. Two major active fractions (P2‐3‐2‐2‐2 and P2‐3‐2‐2‐3) were found to significantly inhibit NO production at 0.156 mg/mL (p < 0.01). In addition, three chemical components (formononetin, calycosin and astragaloside IV) were successfully isolated from P2‐3‐2‐2‐3. Only formononetin could significantly inhibit NO production (p < 0.01), whereas the other two components had no significant effects at concentrations ranging from 0.039 to 0.156 mg/mL. In conclusion, two major anti‐inflammatory active fractions that may enhance wound healing were identified, and formononetin was one of the active ingredients in the active fractions. Copyright © 2012 John Wiley & Sons, Ltd.