Simultaneous Determination of Saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by High Performance Liquid Chromatography

To establish a method for determining five saponins(notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ammonium glycyrrhizinate) in Glycyrrhizae, Notoginseng and Ginseng, the high performance liquid chromatography with diode array detector(HPLC-DAD) method was applied to an Inertsil ODS-SP column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.05% phosphoric acid in a gradient elution manner. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and the detection wavelengths were 203 nm and 237 nm, respectively. The linear ranges were 0.700,0—7.000,0 μg for R1(r=1.000,0), 0.751,1— 7.511,4 μg for Rg1(r=1.000,0), 0.677,2—6.771,6 μg for Re(r=1.000,0), 0.733,9—7.339,1 μg for Rb1(r= 1.000,0), and 0.540,0—5.399,8 μg for ammonium glycyrrhizinate(r=0.999,9), respectively. In addition, their average recoveries were 100.28%, 105.83%, 104.09%, 99.36% and 98.54%, respectively. The relative standard deviations(RSDs) of precision, reproducibility and recovery were all less than 1.5%. The results indicate that the method is simple, accurate and reproducible so that it can be used for the simultaneous determination of the five saponins in Chinese patent medicines containing the three kinds of herbs.

Clinical Application of Radix Notoginseng and Its Homology of Medicine and Food

Clinical application of Radix Notoginseng in the last ten years is summarized,such as Radix Notoginseng powder,compatibility of Radix Notoginseng with other traditional Chinese medicines,and related preparations of Radix Notoginseng,as well as application of Radix Notoginseng in homology of medicine and food,such as preventing and controlling gynecological diseases,rheumatoid arthritis,hypertension,gastric cancer,arthritis,and anemia.

Network pharmacology-based strategy for predicting therapy targets of Sanqi and Huangjing in diabetes mellitus

BACKGROUND A comprehensive literature search shows that Sanqi and Huangjing(SQHJ)can improve diabetes treatment in vivo and in vitro,respectively.However,the combined effects of SQHJ on diabetes mellitus(DM)are still unclear.AIM To explore the potential mechanism of Panax notoginseng(Sanqi in Chinese)and Polygonati Rhizoma(Huangjing in Chinese)for the treatment of DM using network pharmacology.METHODS The active components of SQHJ and targets were predicted and screened by network pharmacology through oral bioavailability and drug-likeness filtration using the Traditional Chinese Medicine Systems Pharmacology Analysis Platform database.The potential targets for the treatment of DM were identified according to the DisGeNET database.A comparative analysis was performed to investigate the overlapping genes between active component targets and DM treatmentrelated targets.We constructed networks of the active component-target and target pathways of SQHJ using Cytoscape software and then analyzed the gene functions.Using the STRING database to perform an interaction analysis among overlapping genes and a topological analysis,the interactions between potential targets were identified.Gene Ontology(GO)function analyses and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted in DAVID.RESULTS We screened 18 active components from 157 SQHJ components,187 potential targets for active components and 115 overlapping genes for active components and DM.The network pharmacology analysis revealed that quercetin,beta-sitosterol,baicalein,etc.were the major active components.The mechanism underlying the SQHJ intervention effects in DM may involve nine core targets(TP53,AKT1,CASP3,TNF,interleukin-6,PTGS2,MMP9,JUN,and MAPK1).The screening and enrichment analysis revealed that the treatment of DM using SQHJ primarily involved 16 GO enriched terms and 13 related pathways.CONCLUSION SQHJ treatment for DM targets TP53,AKT1,CASP3,and TNF and participates in pathways in leishmaniasis and cancer.

从纤维化探讨血竭、三七“以通求固”治疗子宫腺肌病的机制

子宫腺肌病为育龄期女性常见病、多发病之一,月经量多、痛经为其主要临床表现,进行性增大的子宫和月经过多最终导致重度贫血,严重影响女性身心健康。临床运用发现蔡氏妇科“以通求固”思想治疗子宫腺肌病,代表药物为血竭、三七,明显改善患者月经量多、痛经症状。现将现代医学研究与中医理论相结合,尝试从纤维化角度探索“以通求固”思想治疗子宫腺肌病作用机制,为中医药治疗子宫腺肌病提供新思路。

三七药对的研究进展

三七作为临床常用中药,在临床应用中有多种药对形式。围绕近年来基于网络药理学的三七药对、传统三七药对及三七药对配伍前后化学成分变化的实验研究进行归纳和总结,系统阐释三七药对在临床应用的作用机制和科学内涵,为进一步研究三七药对使用规律、配伍理论、临床应用等方面提供新的思路。

Advances in Researches of Extraction Processes of Total Saponins of Pnanx Notoginseng

Traditional extraction methods of total saponins of Panax notoginseng include cold soaking method,water decoction method,alcohol reflux method,percolation method,macroporous resin adsorption method,and accelerated solvent extraction( ASE) method. Modern extraction methods include ultrasonic extraction,microwave assisted extraction,supercritical CO_2 extraction,microbial fermentation assisted extraction,neural network model optimized extraction method,and multi-stage countercurrent extraction method. This paper discussed principles of these methods and compared their advantages and disadvantages.

基于网络药理学与动物实验探究人参—黄连—三七药串治疗糖尿病肾病的作用机制

目的 应用网络药理学的研究方法筛选人参—黄连—三七药串治疗糖尿病肾病的作用靶点及相关信号通路,通过动物实验明确其疗效与作用机制。方法 运用Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)数据库检索获取人参—黄连—三七的主要化学成分以及相关作用靶点;使用DisGeNET筛选胰岛素抵抗与糖尿病肾病的相关靶标基因;通过Venn软件筛选出药物与疾病的共同作用靶点;构建出疾病—靶点—成分—药物网络;使用STRING数据库完成蛋白质—蛋白质相互作用网络(protein-protein interaction networks, PPI)的构建;通过基因本体(gene ontology, GO)富集分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)通路富集分析对有效作用靶点进行分析。以db/db小鼠作为动物模型,于给药第4、第8周后留取各组小鼠8小时尿液,检测各组小鼠尿微量白蛋白排泄水平;采用蛋白免疫印迹法法对网络药理学的研究预测的关键通路及靶点进行进一步的实验验证。结果 本研究共筛选得到人参—黄连—三七药串的人参皂苷Rh2、人参皂苷F2、槲皮素、小檗碱等39种有效成分;获得人参—黄连—三七药串治疗糖尿病肾病潜在靶点18个,其中PTEN、CCL2、IL6、TNF、HIF1A等11个与其他靶点相互作用较强,可能在网络中起到关键作用;通过GO分析和KEGG分析分别获得2595个条目和107条信号通路,其中包括磷脂酰肌醇-3-激酶丝氨酸/苏氨酸特异性蛋白激酶(phosphatidylinositol-3-kinase/protein kinase B,PI3K/Akt)信号通路、晚期糖基化终末产物—晚期糖基化终末产物受体信号通路、低氧诱导因子-1信号通路、细胞迁移的正调控、细胞因子活性、细胞因子受体结合、血小板α颗粒等。动物实验显示,在给药第4周及第8周后,中药组小鼠尿微量白蛋白排泄水平均较模型组下降(P<0.05)。对于PI3K/Akt信号通路的相关蛋白检测结果表明,中药组小鼠肾脏PI3K、p-Akt表达水平较模型组明显下降(P<0.05),磷酸酶张力蛋白同源物(phosphatase and tensin homolog deleted on chromosome ten, PTEN)水平较模型组升高(P<0.05)。结论 人参—黄连—三七药串能够有效改善db/db小鼠糖尿病肾病引起的蛋白尿,其作用机制可能与激活PTEN抑制PI3K/Akt信号通路有关。

Box-Behnken响应面法优化复合益生菌发酵三七药渣工艺及体外抗氧化活性研究

目的以三七药渣为原料,通过益生菌发酵比较不同益生菌以及复合益生菌对各指标成分影响的差异,探究最佳发酵工艺。方法采用单因素、Box-Behnken响应面法优化发酵工艺,并通过体外抗氧化实验评价发酵产物的抗氧化能力。结果最佳发酵工艺为有氧发酵48 h、厌氧发酵36 h,嗜热链球菌、两歧双歧杆菌、嗜酸乳杆菌的比例为2∶3∶1,料液比0.14 g·mL^(-1),接菌量5%,温度33℃。在最佳发酵工艺条件下,三七药渣中性多糖、酸性多糖、总黄酮的含量分别提高了105.64%、96.98%、123.83%,相较于单菌发酵均显著升高。发酵产物清除DPPH、ABTS自由基的IC 50值分别为1.774、3.065 mg·mL^(-1),还原Fe^(3+)的能力为0.138 mmol FeSO_(4)·g^(-1),较未发酵药渣显著增强。结论最佳发酵工艺能显著提高三七药渣中各指标成分含量,显著增强其抗氧化能力,相较单菌发酵各指标成分含量均显著提高,提示上述益生菌之间没有明显的拮抗作用。研究结果为三七药渣的资源化利用提供了科学依据和数据支撑。

三七治疗动脉粥样硬化药理机制研究进展

动脉粥样硬化(atherosclerosis,AS)是引发心脑血管系统疾病的主要原因之一,是动脉内膜异常沉积日久所致的慢性炎症性疾病,病机复杂、治疗周期长久,是临床需要重点解决的重点疾病之一。其病理机制涉及血管平滑肌细胞增殖、内皮损伤、血管内血栓形成、炎症反应、脂质代谢异常、氧化应激加剧、自噬稳态失衡等,而当前对中药三七有效成分是近年来研究的热点和重点,其富含多种皂苷类和非皂苷类有效化学成分,能够综合调理多方面的通路、分子机制,达到减轻炎症反应和内皮损伤,恢复正常脂质代谢、重新调整自噬稳态等的平衡,抑制血管内血栓形成、血管平滑肌细胞增殖,并最终达到预防和治疗AS的目的。鉴于三七在抗AS中的重要作用,该文就三七及其有效成分对AS保护作用的体内外等研究进行综述,为中医药新产品研究开发及临床应用辨证治疗用药提供参考和依据。

基于网络药理学和分子对接探讨三七-骨碎补治疗股骨头坏死的作用机制

目的 基于网络药理学和分子对接探究“三七-骨碎补”药对治疗股骨头坏死的作用机制。方法 利用中药系统药理学数据库与分析平台(TCMSP)对“三七-骨碎补”药对的活性成分及成分靶点进行采集、筛选,通过CNKI、PubMed数据库对其进行补充,并利用UNIPORT数据库对靶点进行转换;通过GeneCards、TTD和OMIM数据库收集股骨头坏死的作用靶点,取得交集靶点后导入STRING数据库,生成PPI网络,并通过Cytoscape 3.9.0软件构建中药-活性成分-靶点网络;通过DAVID数据库对核心靶点进行GO功能富集分析与KEGG通路富集分析。结果 共得到“三七-骨碎补”药对的20个活性成分和229个潜在靶点,1386个股骨头坏死的靶点。GO富集分析主要包括炎症反应、凋亡过程、调节血管生成、与雌性激素响应等功能;KEGG富集分析主要包括脂质与动脉硬化、白细胞介素-17信号通路、类风湿性关节炎等信号通路。结论 “三七-骨碎补”药对治疗股骨头坏死具有成分多、靶点广、通路复杂的特点,可能通过IL6、TNF、TP53等靶点以及脂质代谢、白细胞介素-17等信号通路共同发挥治疗股骨头坏死的作用。

试论三七补血

三七首载于《跌损妙方》,《本草纲目》云三七主治“吐血、咳血不止”“无名痈肿,疼痛不止”,后世本草多奉《本草纲目》所载为圭臬,言三七散瘀止血,消肿定痛,为活血化瘀之神药,直至《滇南本草》载三七“生用破血,炙用补血”,三七始有补血之说。现代研究发现熟三七的补血功效优于生三七,可以缓解血虚患者的临床症状,改善外周血象指标,通过保护造血器官、促进造血干/祖细胞的增殖、提高造血相关因子水平、改善造血微环境及代谢等作用发挥补血的作用。通过对三七的文献记载和现代研究的系统梳理,可为今后三七补血功效的临床应用及现代研究提供理论依据与参考。

三七总皂苷对脑缺血再灌注后神经功能恢复的影响

目的探究三七总皂苷对脑缺血再灌注后神经功能恢复的影响。方法培养30只SPF级雄性SD大鼠,构建脑缺血再灌注模型。将大鼠分为假手术组、模型组和三七总皂苷组,每组10只大鼠。检测各组大鼠脑梗面积、脑含水量。Longa评分评价大鼠神经功能缺损情况。水迷宫实验检测大鼠神经功能。检测各组大鼠血清中氧化应激指标超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、谷光甘肽(glutathione,GSH)含量和炎症因子指标肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-10(interleukin-10,IL-10)含量。Tunel染色检测各组大鼠海马神经元细胞凋亡情况。结果和假手术组大鼠相比,模型组大鼠脑梗面积、脑含水量、神经功能缺损评分、学习潜伏期和记忆潜伏期显著升高,SOD、GSH、IL-10含量显著降低,MDA、TNF-α、IL-1β含量显著升高,海马神经元凋亡比例显著升高(P<0.05)。和模型组大鼠相比,三七总皂苷组大鼠脑梗面积、脑含水量、神经功能缺损评分、学习潜伏期和记忆潜伏期显著降低,SOD、GSH、IL-10含量显著升高,MDA、TNF-α、IL-1β含量显著降低,海马神经元凋亡比例显著降低(P<0.05)。结论三七总皂苷可能通过抑制大鼠炎症反应和氧化应激,减少海马神经元凋亡,促进脑缺血再灌注后神经功能恢复。

三七皂苷成分及临床药理作用研究进展

三七作为传统中草药现已受到医药界的共同认可和关注,现代植物化学研究表明三七皂苷是三七的主要化学成分。现代药理研究及临床应用发现,其具有抗癌、抗氧化及治疗心血管疾病等作用。综述三七主要化学成分和药理作用的研究进展,以期为三七的临床应用及后期研究提供帮助。

红三七炮制工艺优化及其补血活性研究

目的优化红三七炮制工艺,并评价其补血活性。方法在单因素试验基础上,以蒸制温度、蒸制时间、烘干温度、烘干时间为影响因素,三七皂苷R1,人参皂苷Rg_(1)、Rb_(1)、Rk_(3)、Rh_(4),20(R)-人参皂苷Rg_(3)总含量为评价指标,Box-Behnken响应面法优化炮制工艺。建立乙酰苯肼(APH)-环磷酰胺(CTX)联合诱导的小鼠贫血模型,比较红三七、熟三七补血活性差异。结果最佳条件为蒸制温度130℃,蒸制时间4 h,烘干温度60℃,烘干时间48 h,总皂苷含量为8.326%,RSD为0.087%。药理活性表明,不同炮制工艺对三七补血活性有影响,红三七的补血活性优于熟三七。结论该方法稳定可行,可用于红三七的规范生产。

冠心生脉丸质量标准提升

目的提升冠心生脉丸质量标准。方法TLC法定性鉴别人参和三七,分析采用硅胶G薄层板,以三氯甲烷-甲醇-水(13∶7∶2)10℃以下放置的下层溶液为展开剂,10%硫酸乙醇溶液为显色剂。HPLC法测定人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd的含量,分析采用Thermo Accucore-C18色谱柱(4.6 mm×150 mm,2.6μm);流动相乙腈-水,梯度洗脱;体积流量0.8 mL/min;柱温20℃;检测波长203 nm。结果TLC斑点清晰,阴性无干扰。4种成分在各自范围内线性关系良好(R^(2)≥0.9999),平均加样回收率91.21%~106.86%,RSD 0.68%~1.43%。结论该方法专属性、重复性良好,可为冠心生脉丸质量控制提供参考。

基于激光诱导击穿光谱技术的三七产地及道地研究

目的从元素角度出发,采用激光诱导击穿光谱(laser-induced breakdown spectroscopy,LIBS)结合化学计量学方法研究三七的道地性及产地鉴别。方法采集云南文山三七和曲靖三七的LIBS光谱数据,进行数据归一化处理,利用偏最小二乘判别分析(PLS-DA)方法判别三七产地,并采用VIP值筛选出对产地判别模型的贡献大的元素。结果基于LIBS技术和PLS-DA方法可以有效地进行云南文山三七和曲靖三七的产地判别。以450~840 nm全光谱为输入变量,PLS-DA判别模型训练集的敏感性、特异性和总判正率均为100%,预测集敏感性、特异性和总判正率分别为90.9%、90%和95%;以VIP值筛选的特征变量作为输入变量,PLS-DA判别分析模型的性能良好,训练集和预测集的敏感性、特异性和总判正率均为100%。表明VIP值筛选的特征变量可代表三七的产地信息。贡献度较大的元素分别为:Si、Mg、Ca、Na、K等。结论LIBS技术结合多变量分析方法能够快速实现云南文山三七和曲靖三七的道地性研究。所建立的方法具有分析快速、近似无损、样品前处理简单等特点,为中药产地和道地性研究提供新思路。

基于网络药理学探究三七叶对糖尿病大鼠的治疗机制

目的 利用网络药理学和动物实验,研究三七叶抗糖尿病的主要活性成分和潜在作用机制。方法 基于Pubchem、PharmMapper、OMIM等数据库筛选三七叶与糖尿病共同的潜在靶点之后,再利用STRING数据库和Cytoscape软件构建蛋白相互作用网络图,对其进行拓扑学分析和聚类分析获得核心靶点,并进行GO和KEGG富集分析筛选药物作用于疾病的可能机制。随后进行动物实验验证三七叶的抗糖尿病作用。结果 预测结果共获得三七叶与糖尿病的共同靶点50个,核心靶点有3个:ANXA5、CFB、GSTP1,共涉及生物过程986种,细胞组成15种,分子功能74种,信号通路101条。动物实验表明三七叶可降低糖尿病大鼠的血糖,减少与糖尿病肾病相关的p38 MAPK和TGF-β1 mRNA的表达,同时也可以调节氧化应激和血脂水平,减少动脉粥样硬化。结论 三七叶是通过多途径多靶点共同发挥抗糖尿病的作用,其作用机制可能与减少氧化应激、调节细胞因子、减少动脉粥样硬化和抗糖尿病肾病肾纤维化等有关。

三七对下腔静脉结扎DVT大鼠模型D-二聚体、cTn-I、BNP的影响

目的观察三七对下腔静脉结扎深静脉血栓(DVT)大鼠D-二聚体、心肌肌钙蛋白I(cTn-I)、血B型钠尿肽(BNP)的影响。方法将20只大鼠随机分为三七组、DVT组、依诺肝素组和假手术组,每组各5只。除假手术组外均行开腹结扎下腔静脉造模。下腔静脉结扎造模成功后48 h处死大鼠,取游离左肾静脉和下腔静脉汇合处及向下1 cm血管进行苏木素-伊红染色,于显微镜下观察,并检测血管D-二聚体、cTn-I、BNP含量。结果四组大鼠血浆cTn-I、D-二聚体、BNP含量比较,差异均有统计学意义(F分别=26.56、9.86、7.34,P均<0.05)。DVT组大鼠血浆中cTn-I、D-二聚体、BNP含量高于假手术组,差异有统计学意义(t分别=11.40、6.67、8.92,P均<0.05)。三七组、依诺肝素组cTn-I含量明显低于DVT组,差异均有统计学意义(t分别=2.62、3.50,P均<0.05)。依诺肝素组D-二聚体含量明显低于DVT组,差异有统计学意义(t=3.66,P<0.05)。结论三七和依诺肝素对于DVT大鼠模型均有一定治疗作用。

Blood circulation activating effect of Sanqi(Radix Notoginseng) on venous thromboembolism rats

OBJECTIVE:To explore the blood circulation activating effect and mechanism of Sanqi(Radix Notoginseng)in vivo,using a venous thromboembolism(VTE)rat model.METHODS:We established the VTE rat model,and then intervened with low molecular weight hepa-rin(LMWH),as well as low,medium and high doses of Sanqi(Radix Notoginseng),to observe the blood circulation activating effect of Sanqi(Radix Notoginseng)on VTE rats.RESULTS:After the treatment with high concentrations of Sanqi(Radix Notoginseng),the pulmonary thromboembolism was alleviated,and the lower limb thrombosis was markedly improved.Moreover,the expression quantities of plasma activated partial thromboplastin time,prothrombin time and D-dimer,as well as endothelin,von Willebrand factor,and plasminogen activator inhibitor-1 in thrombosis segment tissues were markedly down-regulated;while those of nitric oxide and tissue-type plasminogen activator were up-regulated.After low and medium concentration Sanqi(Radix Notoginseng)treatment,no obvious improvement was observed in each index.Moreover,the high concentration Sanqi(Radix Notoginseng)showed comparable efficacy to the positive drug LMWH.CONCLUSION:This data suggests that high concentration of Sanqi(Radix Notoginseng)is effective in preventing and treating VTE.

Efficacy of Sanqi(Radix Notoginseng)in treating cerebral hemorrhage in rats with traumatic brain injury

OBJECTIVE:To evaluate the protective efficacy of Sanqi(Radix Notoginseng)on cerebral hemorrhage in a rat model of traumatic brain injury(TBI)by investigating plasminogen activator inhibitor-1(PAI-1),tissue-type plasminogen activator(t-PA),nuclear factor-κB(NF-κB,p-p65),nitric oxide(NO),endothelin(ET),cluster differentiation(CD61CD62),and coagulation.METHODS:The free-fall method was used to create a rat model of TBI.Forty-eight rats were randomly divided into six groups:the blank group,sham group,model group,low-dose Sanqi(Radix Notoginseng)group,middle-dose Sanqi(Radix Notoginseng)group,and high-dose Sanqi(Radix Notoginseng)group.At 24 h after the model was created,we investigated brain MRI,brain tissue morphology using HE staining,flow cytometry,and immunohistochemical changes.RESULTS:Cerebral hemorrhage was aggravated in TBI rats(observed in brain specimens,brain MRI,and brain tissue HE).Cerebral immunohistochemistry results demonstrated that the expression of t-PA,PAI-1 and p-p65 increased significantly in TBI rats,while t-PA/PAI-1 had a significant decrease.In addition,CD61CD62,D2D,and ET were significantly increased in TBI rats,and PT and APTT were significantly prolonged;in contrast,NO was significantly decreased.Sanqi(Radix Notoginseng)decreased cerebral hemorrhage in TBI rats(observed in brain MRI and brain tissue HE),and increased t-PA/PAI-1,CD61CD62 significantly.It also significantly decreased the expression of t-PA,PAI-1,and p-p65 in brain immunohistochemistry and significantly decreased PT,APTT,D2D,and ET.However,there were no differences in NO between the model group and the Sanqi(Radix Notoginseng)group.CONCLUSION:Sanqi(Radix Notoginseng)can decrease the expression of p-p65,increase t-PA/PAI-1,and stem traumatic intracranial hemorrhage in a TBI rat model.