三七皂苷PNS-5的溶血性及生物免疫活性研究

目的:研究三七皂苷PNS-5的溶血性及生物免疫活性。方法:常规方法提取PNS,大孔树脂柱层析分离纯化得PNS-5。溶血试验测定红细胞溶血率,脾淋巴细胞增殖反应测定PNS-5的生物免疫活性。结果:PNS-5溶血性较小,其HD50大于8mg/ml;PNS-5(0.1μg/ml)显著促进了ConA和LPS诱导的小鼠脾淋巴细胞增殖反应。结论:PNS-5安全性好,能促进细胞和体液免疫。

基线等比增减设计法优选PNS/TGCO组分对实验性气虚血瘀型冠心病的影响研究

目的:初步探索三七总皂苷(PNS)和山茱萸总苷(TGCO)配伍治疗气虚血瘀型冠心病的作用及特点。方法:采用基线等比增减设计法拟定PNS/TGCO组合配比,观察不同比例组合的有效组分对垂体后叶素所致大鼠急性心肌缺血的影响以及对正常小鼠耐缺氧时间的影响。结果:处方1、2、3及血塞通有一定改善大鼠心肌缺血程度的作用,处方4、5、6与对照组比较无显著性差异;处方3、4、5、6有一定抗小鼠缺氧的作用,处方1、2及血塞通组与对照组比较无显著性差异。结论:处方3既有一定改善大鼠心肌缺血的作用又有一定抗小鼠缺氧的作用,为PNS/TGCO治疗气虚血瘀型冠心病有效组分的最佳配比。

A Novel Hexanordammarane Glycoside from the Roots of Panax notoginseng

Notoginsenoside R-111, a novel hexanordammarane glycoside was isolated from the roots of Panax nototginseng, as a minor constituent. Its structure was determined as 6-O-(beta -D-glucopyranosyl)-3 beta ,6 alpha ,12 beta -trihydroxy-22,23,24,25,26,27-hexanordammaran-20-one(1), by means of spectroscopic methods.

Two New Dammarane Glycosides from the Acid Hydrolysis Product of Panax Notoginseng

Two new dammarane glycosides named notoginsenoside T-1 and T-2 were isolated from the mild acid hydrolysis products of the root saponins of Panax notoginseng. On the basis of spectroscopic evidences, their structures were elucidated to be 6-O-beta -D-glucopyranosyl-24(25)-epoxy-3 beta ,6 alpha ,12 beta ,23 xi -tetrahydroxydammar-20(22)(E)-ene 1 and 6-O-beta -D-glucopyranosyl-24(25)-epoxy-23 xi -methoxyl-3 beta ,6 alpha ,12 beta -trihydroxydamm-ar-20(22)(E)-ene 2, respectively.

Dammarane-type saponins from steamed leaves of Panax notoginseng

Four new dammarane-type triterpenoid saponins,namely notoginsenosides SFt1-SFt4(1-4)were isolated from the steamed leaves of Panax notoginseng(Burk.)F.H.Chen(Araliaceae),together with 17 known saponins.Their structures were established on the basis of detailed spectroscopic analyses and acidic hydrolysis.The known ginsenosides Rk2 and Rh3 were obtained from P.notoginseng for the first time.All of these new saponins showed no in vitro cytotoxicity against five human cancer cell lines(HL-60,SMMC-7712,A-549,MCF-7,and SW480).

Effects of Panax notoginseng saponins on apoptosis induced by hydrogen peroxide in cultured rabbit bone marrow stromal cells via altering the oxidative stress level and down-regulating caspase-3

Objective： To investigate the effects of Panax notoginseng saponins （PNS） on hydrogen peroxide （H2O2）-induced apoptosis in cultured rabbit bone marrow stromal cells （BMSCs）. Methods： The effects of different concentrations of PNS on proliferation and early osteoblast differentiation of BMSCs were determined by the MTT assay and an alkaline phosphatase （ALP） assay. An optimal effective concentration of PNS was determined and used in subsequent experiments. The cultured BMSCs were divided into three groups： untreated control, H2O2 treated, and PNS pretreatment of H2O2 treated. The oxidative stress level was assessed by superoxide dismutase （SOD） and malondialdehyde （MDA） assays. Flow cytometry was used to determine BMSC apoptosis by staining with annexinV-FITC/propidium iodide （PI）. The activity of caspase-3 enzyme was measured by spectrofluorometry. Results： PNS （0.1g/L） significantly increased both BMSC proliferation rate and ALP activity, while it decreased the indicators of oxidative stress, caspase-3 activity, and the apoptosis rate of BMSCs induced by H2O2.. Conclusion： PNS, acting as a biological antioxidant, had a protective effect on H2O2-induced apoptosis in cultured rabbit BMSCs by decreasing oxidative stress and down-regulating caspase-3.

Panax notogiseng saponin inhibits ischemia-induced apoptosis by activating PI3K/Akt signal pathway in cardiomyocytes

The panax notoginseng saponin(PNS) had been clinically used for the treatment of cardiovascular diseases and stroke in China.It had been demonstrated that PNS could protect cardiomyocytes from injury induced by ischemi- a,but the underlying molecular mechanisms of this protective effect were still unclear.This study was aimed to investigate the protective effect and molecular mechanisms of PNS on apoptosis in H9c2 cells in vitro and rat myocardial ischemia injury model in vivo.Annexin-V/PI assay shew that PNS could protect H9c2 cells from apoptosis induced by serum, glucose and oxygen deprivation(SGOD) in a dose-dependent manner.However,the anti-apoptotic effect of PNS was reversed by LY294002,a specific PI3K inhibitor.This antiapoptotic effect of PNS was confirmed by JC-1,a specific probe of mitochondrial membrane potential staining.PNS could significantly increase phos-Akt in H9c2 cells by Western blot assays and its effect could be inhibited by LY294002.Furthermore,PNS could improve ischemic-induced left ventricular function as reflected by EF,LVDd and LVDs.PNS could also inhibited cellular apoptosis in myocardial tissues in ischemic rats by TUNEL assay.PNS administration also increased the expression of phos-Akt in rat ischemic myocardial tissues.These results suggested that PNS could protect myocardial cells from apoptosis induced by ischemia in vitro model and in vivo model through activating-PI3K/Akt signal pathway which may be meaningful for further understanding the molecular mechanisms of cardiac protection of PNS.And the results might be useful in treatment of myocardial ischemia in future.

EFFECTS OF TOTAL SAPONINS OF PANAX NOTOGINSENG AND LIGUSTRAZINE ON THE PROLIFERATION OF CEREBRAL MICROVASCULAR ENDOTHELIAL CELLS OF RATS

Objective To investigate the effects of Total Saponins of Panax notoginseng(PNS) and Liguastrazine(LIT) on the proliferation of cultured cerebral microvascular endothelial cells. Methods The inverted microscope was used to observe endothelial cells and immunochemical methods was also used to detect FVIII-related antigens so as to observe endothelial cells. PNS or LIT in concentrations 0.5?g·L -1, 1.0?g·L -1 and 2.0?g·L -1 were used on the cultured cerebral endothelial cells of rats for 24 hours. MTT method was adopted to determine the outcome of endothelial proliferation. Results 1. Immunochemical methods was used to detect FVIII-related antigens. The brownish yellow showed positive, and the observation of the cultured endothelial cells under inverted microscope showed that the cells appeared to be in the morphological form of cobble-stones. 2. PNS in lower concentration (0.5?g·L -1) could facilitate the proliferation of the cells, while 1?g·L -1 and 2?g·L -1 of PNS could inhibit the proliferation of the cells. 0.5?g·L -1 of LIT could facilitate the proliferation of cellswhile LIT of 1?g·L -1 and 2?g·L -1 had no significant effect. Conclusion The two kind of TCM ingredients extracted in lower concentration could facilitate the proliferation of the cells. And, at the same concentration, the inhibition of PNS on the cells is stronger than that of LIT.

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.

三七皂苷R1药理作用机制与神经保护作用研究进展

三七是五加科人参属多年生直立草本植物三七[Panax notoginseng(Burk.)F.H.Chen]干燥根及根茎,作为我国传统中药,具有较好的补益功效,《中药大辞典》有言,三七功用补血,去瘀损,止血衄,能通能补,功效最良,是方药中之最珍贵者。生吃去瘀生新,消肿定痛,止血不留瘀,行血不伤新;熟服可补益健体。三七皂苷R1是三七总皂苷中特征化合物,近些年来,我国老龄化程度不断加剧,神经系统疾病发生率逐年增高,同时,对三七皂苷R1治疗神经疾病的报道增多,其发挥神经保护作用有确切的疗效。该文旨在对三七皂苷R1治疗神经疾病以及作用机制展开综述,为临床用药与新药开发提供一定的理论依据。

三七对耐甲氧西林金黄色葡萄球菌细胞壁的作用及机制

目的 探究三七(PN)对耐甲氧西林金黄色葡萄球菌(MRSA)细胞壁的作用及机制。方法 实验分对照组(Control组)和PN组,Control组MRSA未干预,PN组MRSA被2 mg/mLPN干预24 h。酶联免疫吸附试验(ELISA)检测PN对MRSA细胞壁肽聚糖(PGN)和脂磷壁酸(LTA)的影响;透射电镜(TEM)观察MRSA细胞壁厚度;转录组测序技术(RNA-Seq)筛选PN干预MRSA的药效相关基因,通过基因本体(GO)与京都基因与基因组百科全书(KEGG)对其进行富集分析;实时荧光定量PCR技术验证PN干预MRSA的细胞壁相关基因。结果 与Control组比较,PN组PGN、LTA降低(P<0.05);PN组处于分裂期的细菌数量增多,体积变大,细菌分裂隔膜模糊,细胞壁厚度薄(P<0.05)。2组间有552个差异基因,相较于Control组,PN组285个基因上调、267个基因下调;差异基因涉及的生物过程(BP)主要富集在氧化还原、前体代谢产物和能量产生、细胞氨基酸分解代谢、有氧呼吸过程,细胞组分(CC)主要富集在细胞解剖实体、细胞器官、非膜界细胞器,分子功能(MF)主要富集在氧化还原酶活性、结构分子活性、电子转移活性;差异基因共富集85条信号通路,其中包括脂肪酸、赖氨酸降解;差异基因MraY、MurD、MurC、FemA、FemB、sgtB与MRSA细胞壁相关。与Control组比较,PN组MraY、MurD、FemA、FemB表达降低(P<0.05),MurC、sgtB表达升高(P<0.05)。结论 PN可抑制MRSA细胞壁,与降低细胞壁相关基因表达和促进脂肪酸、赖氨酸降解等有关。

Pharmacokinetics and brain distribution differences of baicalin in rat underlying the effect of Panax notoginsenosides after intravenous administration

Baicalin(BA) is the most well-known flavonoid present in Radix Scutellariae. The aim of this study was to explore whether the pharmacokinetic behavior of BA in rat brain can be affected by Panax notoginsenosides(PNS), and to assess the possible mechanism for the observed effects. Specific HPLC and HPLC/MS/MS methods were developed and validated for the determination of BA in the rat plasma and brain using carbamazepine as an internal standard. BA was found to enter rat brain quickly after a single intravenous dose. When co-administered with PNS, clearance(CL) of BA from rat plasma decreased by 50.00%, while the area under the curve AUC0-t and AUC0-∞ increased 94.69% and 87.68%, respectively. On the other hand, some pharmacokinetic parameters of BA in rat brain had obvious differences after PNS was administered, such as an increase in Tmax from 5 min to 15 min, an increase in AUC0-t and AUC0-∞ by 42.75% and 29.39%, respectively, as well as a decrease in CL by 27.95%. Together, these results indicate that PNS can decrease the elimination rate of BA from rat plasma, promote the penetration of BA into rat brain, increase the concentration and slow down the elimination of BA from rat brain. The data provide important information that compatibility with PNS can promote the consequent effects of BA for the treatment of encephalopathy.

三七总皂苷通过p38 MAPK通路抑制内毒素诱导的小胶质细胞活化

目的:探讨三七总皂苷(PNS)通过p38丝裂原活化蛋白激酶(p38 MAPK)通路对脂多糖(LPS)诱导活化的BV2小胶质细胞中肿瘤坏死因子-α(TNF-α)表达的影响。方法:将BV2小胶质细胞分为空白对照组(Control)、LPS激活组和LPS+三七总皂苷干预组(LPS+PNS)。CCK-8法检测BV2小胶质细胞的活力,确定最适合的药物干预浓度。利用Western Blot和免疫荧光检测BV2小胶质细胞中p38 MAPK和TNF-α的表达及p38 MAPK磷酸化水平(p-p38 MAPK)变化。结果:与空白对照组相比,PNS对BV2小胶质细胞的细胞活力无显著差异,最终选定100 mg/L作为药物干预浓度。Western Blot、免疫荧光结果提示,LPS激活后,BV2小胶质细胞中TNF-α的表达显著升高,p38 MAPK磷酸化水平增加(P<0.05);PNS干预后,与LPS激活组相比,TNF-α表达显著下降,p38 MAPK磷酸化水平降低(P<0.05)。使用p38 MAPK通路抑制剂(SB203580)作用后,PNS联合SB203580组(LPS+PNS+SB203580)中,TNF-α表达及p38 MAPK磷酸化水平变化与LPS+PNS组相比无显著差异(P>0.05)。此外,p38 MAPK在各组的变化无显著性差异(P>0.05)。结论:PNS可能通过p38 MAPK通路抑制活化的BV2小胶质细胞分泌的炎性因子TNF-α的表达。

基于网络药理学及分子对接探讨三七总皂苷治疗脓毒症的作用机制

目的基于网络药理学与分子对接技术探讨三七总皂苷治疗脓毒症的作用机制。方法通过检索TCMSP、Swiss Target Prediciton和PharmMapper数据库得到三七总皂苷的作用靶点;在Genecard、Drugbank、Disgenet和OMIM数据库检索脓毒症的疾病相关靶点;将筛选出的三七总皂苷作用靶点与脓毒症疾病相关靶点取交集,作为三七总皂苷治疗脓毒症的潜在作用靶点。通过STRING数据库构建潜在作用靶点的蛋白互作(PPI)网络,并筛选关键靶点;对潜在作用靶点进行GO功能及KEGG通路富集分析,构建药物-疾病-靶点-通路网络;利用AutoDock Vina软件对三七总皂苷5种单体皂苷成分与三七总皂苷治疗脓毒症的关键靶点进行分子对接验证。结果获得三七总皂苷治疗脓毒症的潜在作用靶点206个;筛选得到AKT1、TP53、SRC、STAT3、JUN、TNF、IL6、MAPK1、PIK3R1和IL1B等关键靶点。潜在作用靶点GO功能富集分析得到2548项生物过程(BP)条目,174项分子功能(MF)条目,47项细胞组分(CC)条目;KEGG通路富集分析共得到171条信号通路。三七总皂苷主要活性成分三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd与关键靶点AKT1、TP53、STAT3、SRC、JUN具有较强的结合活性。结论三七总皂苷可能通过AKT1、TP53、SRC、STAT3、TNF等关键靶点,作用于PI3K-Akt、AGE-RAGE等主要信号通路,进而影响脓毒症的炎症反应、细胞凋亡和凝血反应等生理过程,发挥治疗脓毒症的作用。

三七总皂苷对心梗后心室重构大鼠增强抗氧化与改善心肌形态学作用

为研究三七总皂苷（PNS）对急性心肌梗塞（AMI）后左室重构（LVRM）大鼠自由基损伤和心肌细胞形态学改变的作用，采用结扎大鼠左冠状动脉前降支的方法，建立AMI模型，术后24h后随机分为对照组和实验组，连续4周分别灌胃给予生理盐水和PNS低、中、高剂量，观察PNS和福辛普利对病鼠血清丙二醛（MDA）、c反应蛋白（CRP）、肌红蛋白-I（cTn—I）及肌酸激酶同工酶（CK—MB）、谷胱甘肽过氧化物酶（GSH—Px）、一氧化氮（NO）及心肌细胞形态病理结构和心脏指数改变等的影响。结果发现与对照组比较，PNS与福辛普利均能显著改善病鼠左室心肌细胞形态结构病理改变及心脏指数（P〈0．01），显著降低MDA、CRP、CK—MB与cTn—I、提高GSH—Px活性（P〈0．01—0．05），高剂量PNS可明显降低NO含量（P〈0．05）。可见PNS可通过抑制脂质过氧化反应，减轻病鼠心肌细胞的病理损伤，增强抗氧化，具有抑制心肌肥大与改善心室重构心肌细胞形态学结构作用。

Analysis of the Constituents in the Chinese Drug Notoginseng by Liquid Chromatography-Electrospray Mass Spectrometry

To develop a HPIX-UV-MS method for identifying the constituents in theChinese drug Notoginseng (the root of Panax notoginseng). Methods A Phenomenex Luna C_(18) column(250 mm x 4.6 mm ID, 5 μm) was utilized. Water containing 0.005% formic acid (A) and acetonitrilecontaining 0.005% formic acid (B) were used as gradient eluents. UV spectra were recorded in range195 - 400 nm. Both positive and negative ion ESI modes were used. Results The constituents inNotoginseng were well separated and detected. Fourteen compounds were identified by comparing theirretention time and ESI-MS data with those obtained from the reference compounds. Forty-one compoundswere deduced by data analysis of MS and literature; among them, yesanchinosides-H and -E,chikusetsusaponin-L_5, malonyl-ginsenoside-R_(g_1), the isomers of notoginsenosides-J, -A, -R_1, -G,-R_2, and ginsenoside-Rh_3 were discovered in Notoginseng for the first time. Conclusion Thismethod gives high sensitivity and good separation, and is suitable for identifying the constituentsin Notoginseng. This result is helpful for further phytochemical research on Notoginseng. Based onthis result, further quality control can be studied.

商品人参根的化学成分研究

目的 :分离鉴定人参根的化学成分。方法 :大孔吸附树脂、硅胶柱色谱及HPLC方法分离 ,通过对其进行理化常数和光谱分析方法确定结构。结果 :自商品人参中分离得到 16个皂苷类成分 ,分别鉴定为三七皂苷 R2 (1) ,人参皂苷 Rg2 (2 ) ,2 0 (R) Rg2 (3) ,人参皂苷 Rg1(4) , Rf(5 ) , Re(6 ) , Rd(7) , Rc(8) , Rb1(9) , Rb2 (10 ) , Rb3 (11) , Ra3 (12 ) , Ra2 (13) , Ra1(14 ) ,三七参苷 R4(15 )和人参皂苷 Ro(16 )。结论 :化合物 1为首次自本种植物分离得到。

三七总苷对尿毒血清诱导的人肾小管上皮细胞TGF-β_1、CTGF基因表达和蛋白分泌的影响

目的探讨三七总苷(PNS)对尿毒血清诱导的人肾小管上皮细胞转化生长因子-β1(TGF-β1)、结缔组织生长因子(CTGF)基因表达和蛋白分泌的影响,为临床上应用PNS延缓肾衰竭的进展提供理论依据.方法无菌条件下收集40份尿毒症病人血清和20例正常人血清.应用相差显微镜、扫描电镜结合细胞角蛋白18(CK-18)免疫组化鉴定人肾小管上皮细胞株(HK-2).应用ELISA方法检测HK-2细胞培养上清液TGF-β1蛋白分泌量;Western Blot方法检测CTGF的蛋白表达;RT-PCR方法检测TGF-β1、CTGFmRNA表达.结果 10%尿毒血清组与正常对照组相比,TGF-β1、CTGF基因表达和蛋白分泌量均增高(P<0.01),而PNS 400～800 mg·L-1可恢复尿毒血清所诱导的TGF-β1基因表达(P<0.01),200～800 mg·L-1可恢复尿毒血清诱导的TGF-β1蛋白分泌(P<0.01);200～800 mg·L-1可恢复尿毒血清所诱导的CTGF基因表达和蛋白分泌(P<0.01).结论在体外实验中,PNS可在一定程度上降低尿毒血清诱导的HK-2细胞TGF-β1、CTGF基因表达和蛋白分泌量,减少细胞外基质聚集,从而延缓人肾小管-间质纤维化的进展,有望成为一种能有效延缓肾纤维化进展的中药.

三七总皂苷注射液对大鼠肝CYP450酶的影响

目的:在体酶探针法研究三七皂苷注射液对大鼠肝CYP450酶的影响。方法:连续5周静脉注射给予大鼠20mg·kg-1.d-1的三七总皂苷注射液后分别静注5.0mg/kg的尼莫地平或灌胃20.0mg/kg的华法林钠,HPLC法测定尼莫地平和华法林钠的血浆药物浓度,DAS软件计算药动学参数,评价三七总皂苷注射液对大鼠CYP3A4、CYP2C9和CYP1A2酶的影响,采用双侧t检验方法对数据进行分析。结果:建立并优化了大鼠体内尼莫地平和华法林钠的高效、快速和专属性强的HPLC测定方法,三七总皂苷注射液对尼莫地平和华法林钠在大鼠体内的药动学参数无显著性影响,但长时间给予三七总皂苷注射液后有致尼莫地平AUC升高而清除率下降的趋势,也使华法林钠在大鼠体内的tmax、t1/2、AUC有升高的趋势。结论:三七总皂苷对CYP3A4、CYP2C9和CYP1A2可能有一定的抑制作用,提示我们在临床上与上述酶底物合用时应注意药物相互作用出现的可能性。

三七总皂苷对急性心梗后左室重构大鼠心功能的改善作用

目的研究三七总皂苷(PNS)对急性心肌梗塞(AMI)后左室重构(LVRM)大鼠心功能的作用,并与比索洛尔的疗效作比较。方法除假手术组外采用结扎大鼠左冠状动脉前降支的方法建立AMI模型,术后24h后随机分为正常对照组和实验组,连续4周分别灌胃给予生理盐水2 mg.kg-1、比索洛尔25 mg.kg-1和PNS40、80、120 mg.kg-1,观察PNS和比索洛尔对病鼠室间隔舒张末期厚度(IVSd)、左室舒张末期内径(宽度)(LVIDd)、室间隔收缩末期厚度(IVSs)、左室收缩末期内径(宽度)(LVIDs)、左室后壁舒张末期厚度(LVPWd)、左室后壁收缩末期厚度(LVPWs)、左室射血分数(EF)、左室收缩百分率(FS)、二尖瓣口舒张早期血流速度(MV)、心率(HR)等反映心功能变化指标的影响。结果 PNS高、中剂量组与比索洛尔组的MV、EF均明显上升(P<0.01),中剂量PNS组及比索洛尔组的FS明显上升、LVIDd和LVIDs则减小(P<0.01～0.05)。结论 PNS及比索洛尔均具有改善AMI后LVRM病鼠心功能的作用,可用于治疗心梗后的左室重构。