Gut microbiota-mediated metabolism of Panax notoginseng saponins and its role in pharmacokinetics and pharmacodynamics

Panax notoginseng saponins(PNS)are a class of effective ingredients in Notoginseng Radix et Rhizoma,a well-known herbal medicine called San-Qi in Chinese.After oral administration,PNS inevitably interacts with gut microbiota,and thus affect the pharmacokinetic profiles and pharmacological effects.To date,studies concering gut microbiota-mediated metabolism of PNS have not been reviewed systematically.Herein,we outline the metabolic profiles of Panax notoginseng saponins mediated by gut microbiota,as well as its role in the pharmacokinetics and pharmacodynamics on the basis of reported data.The metabolic pathways of primary saponins are proposed,and step-by-step deglycosylation is found to be the primary degradation pathways of PNS mediated by gut microbiota.Specific microorganisms and enzymes involved in the metabolic processes were summarized.Gut microbiota is deeply involved in the metabolism of PNS,affects the pharmacokinetic profiles,and produces a series of active metabolites.These metabolites were documented to play an essential role in the efficacy of the parent compounds.Future studies should focus on strengthening the real-world evidence,defining the interaction between gut microbiota and PNS,and developing the strategy for modulating gut microbiota to enhance the bioavailability and efficacy of PNS.These information would be useful for further research and clinical application of PNS.

The protective effects of Panax notoginseng in the treatment of inflammatory bowel disease:in silico and in vivo studies

Background:Panax notoginseng(PNE)is a prominent traditional Chinese medicine with extensive beneficial effects on the immune system.However,the precise mechanism of PNE in treating inflammatory bowel disease(IBD)remains unclear.Methods:We first used an extensive metabolomics approach utilizing UPLC-ESI-Q TRAP-MS/MS to identify the metabolite components of PNE aqueous extract.Moreover,the mechanism of PNE in treating IBD was investigated through in silico analysis including RNA-seq analysis,Network pharmacology and Molecular docking.Then a Drosophila toxin-induced intestinal inflammation model was employed to investigate further.Results:A total of 1,543 metabolites of PNE aqueous extract were characterized using UPLC-ESI-Q TRAP-MS/MS.In silico analyses showed that 97 IBD hub targets were targeted by 21 PNE ingredients.Kyoto Encyclopedia of Genes and Genomes results indicated that PNE may play an anti-IBD role through the Mitogen-activated protein kinase(MAPK)signaling pathway and other immune-related signaling pathways.Moreover,11 top hits compounds of PNE show a good affinity binding to IBD targets.The experimental results demonstrated that PNE can effectively improve the survival rate of adult Drosophila while also inhibit the excessive proliferation and differentiation of intestinal stem cells induced by sodium dodecyl sulfate.Furthermore,PNE notably lower the epithelial cell mortality,the accumulation of reactive oxygen species and the activation of oxidative stress-associated jun-Nterminal kinase(JNK)pathway.Conclusion:Our data suggests that PNE aqueous extract has a significant protective impact on the intestinal homeostasis of Drosophila.These findings establish a basis for utilizing PNE in clinical investigations and managing IBD.

High-throughput transcriptional profiling of perturbations by Panax ginseng saponins and Panax notoginseng saponins using TCM-seq

Panax ginseng(PG)and Panax notoginseng(PN)are highly valuable Chinese medicines(CM).Although both CMs have similar active constituents,their clinical applications are clearly different.Over the past decade,RNA sequencing(RNA-seq)analysis has been employed to investigate the molecular mechanisms of extracts or monomers.However,owing to the limited number of samples in standard RNA-seq,few studies have systematically compared the effects of PG and PN spanning multiple conditions at the transcriptomic level.Here,we developed an approach that simultaneously profiles transcriptome changes for multiplexed samples using RNA-seq(TCM-seq),a high-throughput,low-cost workflow to molecularly evaluate CM perturbations.A species-mixing experiment was conducted to illustrate the accuracy of sample multiplexing in TCM-seq.Transcriptomes from repeated samples were used to verify the robustness of TCM-seq.We then focused on the primary active components,Panax notoginseng saponins(PNS)and Panax ginseng saponins(PGS)extracted from PN and PG,respectively.We also characterized the transcriptome changes of 10 cell lines,treated with four different doses of PNS and PGS,using TCM-seq to compare the differences in their perturbing effects on genes,functional pathways,gene modules,and molecular networks.The results of transcriptional data analysis showed that the transcriptional patterns of various cell lines were significantly distinct.PGS exhibited a stronger regulatory effect on genes involved in cardiovascular disease,whereas PNS resulted in a greater coagulation effect on vascular endothelial cells.This study proposes a paradigm to comprehensively explore the differences in mechanisms of action between CMs based on transcriptome readouts.

文山三七(Panax notoginseng)种植区三七与土壤中Pb、Cd、Cu和Zn的分布特征及评价

为了调查文山州三七(Panax notoginseng)主要种植区(丘北县、砚山县、文山县和广南县)土壤和三七Pb、Cd、Cu和Zn含量,通过GPS定位采集三七根区0~15 cm深度土壤样品和三七样品各30个,分析土壤中重金属总量和不同形态含量及三七中对应重金属含量,并进行空间分布分析。结果表明:(1)土壤Pb、Cd、Cu和Zn平均含量分别为55.56、0.36、43.53和119.62 mg·kg^(-1),超标率分别为6.67%、53.3%、13.33%和0。各形态Pb、Cu和Zn含量表现为残渣态>有机物结合态>铁锰氧化物结合态>碳酸盐结合态>可交换态。各形态Cd含量表现为铁锰氧化物结合态>残渣态>碳酸盐结合态>有机物结合态>可交换态。(2)三七和土壤Pb、Cd、Cu和Zn总量和不同形态含量均以丘北县最高,广南县最低。(3)三七根系Pb、Cd、Cu和Zn平均含量分别为2.93、0.35、5.21和11.11mg·kg^(-1),三七植株各部位重金属含量表现为根系>剪口>茎叶>花果。调查区三七和土壤重金属含量存在空间差异,且以土壤Cd污染为主,应采取一定措施降低三七Cd含量。

生物质炭、有机肥和钙镁磷肥对三七(Panax Notoginseng)Cd含量的影响

选择生物质炭、钙镁磷肥、有机肥三种改良剂,在云南三七主产区进行田间试验,比较不同改良剂对降低五加科人参属三七（Panax Notoginseng）Cd含量的效果。结果表明,生物质炭和钙镁磷肥处理均显著降低了三七主根、剪口、茎、叶的Cd含量,降低幅度分别为25.4%~43.6%、40.2%~40.9%、34.3%~51.2%和33.0%~33.5%,且生物质炭、钙镁磷肥处理下三七主根干重较对照分别显著提高48.7%和50.4%;生物质炭和钙镁磷肥处理土壤有效Cd含量分别减少56.1%和58.1%,表明生物质炭和钙镁磷肥能有效降低土壤Cd生物有效性、抑制三七Cd吸收。这与生物质炭和钙镁磷肥处理通过降低土壤酸性、提高土壤CEC及有机质含量有关。有机肥处理三七植株生物量和三七各部位Cd含量与对照相比均无显著差异。此外,生物质炭和钙镁磷肥处理显著降低了三七主根、剪口、茎、叶Cd的富集系数（Accumulation coefficient,AF）,对三七Cd转移系数（Transfer coefficient,TF）影响则不显著,而有机肥处理对三七Cd的AF与TF均无影响;各处理三七须根Cd的AF在2.84~4.64之间,显著高于其他部位,而三七主根、剪口、茎、叶等部位Cd的AF和TF均小于1,表明三七须根对土壤Cd富集能力较强而转移能力较差,Cd易集中于三七地下部,Cd污染土壤中施用生物质炭与钙镁磷肥能有效降低Cd在三七体内的富集。

三七(Panax notoginseng)根残体化感自毒效应研究

为研究三七(Panax notoginseng(Burk)F.H.Chen)采收过程中残留须根的化感自毒效应,采用土培和水培2种方法,按不同比例添加三七须根粉碎物,检测根残体作用下土培三七土壤中皂苷的动态变化及水培三七根部形态结构变化。结果显示,土培条件下,随着须根粉碎物处理时间的延长,土壤中皂苷成分种类增加、总皂苷含量减少。水培条件下,随着处理时间的延长,添加须根粉碎物处理后的三七根尖细胞壁增厚,细胞中出现菌丝体;随着处理时间及处理浓度的增加,细胞开始皱缩甚至破碎,细胞中无完整细胞器结构,三七根部细胞结构差异明显。研究表明三七采收过程中残留在土壤中的须根腐解释放的化学物质可能是导致三七自毒效应的因素之一。

文山三七(Panax notoginseng)种植区土壤As空间分布特征及理化性质对三七As含量的影响

三七（Panax notoginseng（Burk.）F.H.Chen）是云南文山州的道地中药材,文山州由于地壳原始分化成分的作用和含砷矿的开采等原因,使三七种植区存在普遍的土壤砷污染现象。通过野外大田调查和室内分析相结合的方法,研究了云南文山三七主要种植区（文山县、丘北县、砚山县、广南县）土壤总砷和有效砷的空间分布特征和土壤物理化学特征（pH值、有机质含量、阳离子交换量CEC、总P、速效P、土壤质地）,探讨了三七不同部位（茎叶、花果、主根、须根、剪口）As含量的影响因素。将有助于了解文山三七种植区砷污染空间分布特点,寻求三七砷含量的调控途径,对降低三七砷含量和提高三七品质具有一定的实践和理论意义。研究结果表明,（1）土壤有效As和总As平均质量分数分别为0.29 mg·kg-1和28.51mg·kg-1,土壤有效As占总As质量分数的0.06%~2.08%。76.67%的土壤样品总As含量达到国家土壤环境质量的二级标准。土壤有效As和总As含量空间分布表现出西北高,东南低的趋势。（2）三七各部位中As质量分数为0.14~0.32 mg·kg-1,砷含量符合优质三七砷的限量标准（≤1.0 mg·kg-1）,三七各部位对As的富集系数为0.004~0.011,均较低。（3）90%的土壤为酸性土壤,pH值≤6.5;土壤全P质量分数为1.27 g·kg-1,土壤速效P质量分数为66.87 mg·kg-1,均属于丰富水平;土壤有机质质量分数为15.77 g·kg-1,CEC为26.74 cmol·kg-1,属于较高水平;40%的土壤为黏土,43.4%为壤质黏土。（4）土壤总As和土壤有效As含量主要受到随机人为因素的影响,表现出与土壤CEC、pH值和有机质含量之间显著的正相关关系。三七须根中As含量与土壤总As、有效As、土壤黏粒含量、CEC、pH值和总P含量之间具有显著的正相关性。总之,壤质黏土、低pH值和CEC有利于降低三七As含量,三七种植过程中应避免施用含As有机肥和磷肥。研究结果在系统了解文山三七种植区的土壤As空间分布和调控途径方面具有一定的创新价值和意义,特别需要关注是三七产业的发展布局中,应该进行适当的引导和科学的安排三七种植区。

连续2年土壤砷胁迫对三七(Panax notoginseng)细胞膜透性和抗氧化酶活性的影响

通过盆栽试验和室内分析相结合的方法,研究不同砷处理(As5+:0、20、80、140、200和260 mg/kg,采用Na3As O4配制)对三七营养生长期叶片中的细胞膜透性、丙二醛(MDA)含量和抗氧化酶[超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和多酚氧化酶(PPO)]活性的影响,探讨三七细胞膜透性和抗氧化系统酶活性对砷胁迫的响应。结果表明:(1)随着砷质量分数的增加,叶片细胞膜透性和MDA含量逐渐升高;连续2年砷处理为260 mg/kg时,细胞膜透性和MDA含量分别比对照增加160.5%和48.8%;叶片中丙二醛含量与砷质量分数呈极显著正相关关系;(2)第1年砷胁迫,SOD和PPO活性维持稳定,POD活性有所降低;连续2年砷胁迫,SOD活性随着砷质量分数的增加呈现先升高后降低的趋势,且砷处理为260 mg/kg时,酶活性仅为对照的35.7%;POD和PPO活性随砷质量分数的增加而降低;2年砷胁迫CAT活性均随着砷质量分数的增加而增加。总之,在砷胁迫条件下,三七叶片细胞膜受到严重伤害,抗氧化酶系统中POD和PPO活性受到抑制,SOD和CAT是三七叶片中清除过氧化作用的关键酶,连续2年砷胁迫对三七叶片的影响具有一定的累积效应。

Studies on the Isolation, Identification and In Vitro Growth Rates of the Three Pathogenic Fungi from Panax notoginseng Cultivated in Wenshan Eparchy

Objective] The aim of this study was to simultaneously isolate and identify the main pathogenic fungi of the root rot, black spot and round spot from the Panax notoginseng plants cultivated in Wenshan Eparchy of Yunnan Province of China. [Method] The pathogenic fungi were isolated and purified by using potato dextrose agar （PDA） medium. The morphological identification was accomplished first according to the colony forms of the fungi when cultivated in vitro, then accord-ing to the symptom characteristics and colony forms of the re-isolated fungi in the reverse inoculation experiments. The molecular identification was performed accord-ing to the amplification and alignment of the internal transcribed space （ITS） se-quences of the fungi. The increases of the diameters and thickness of the colonies of the fungi cultivated in vitro were employed to indicate the growth rates of the fungi. [Results] The consistency of the colony forms and symptom characteristics and the 96%-99% similarities revealed in the ITS sequence alignments al proved that the main pathogenic fungi of the root rot, black spot and round spot of the P. notoginseng plants raised in Wenshan were Cylindrocarpon didymium, Alternaria panax and Mycocentrospora acerina, respectively. When cultivated in vitro in the same temperature, humidity and il umination, the increases of the colony diameters and thickness of C. didymium were the highest, fol owed by those of A. panax, then those of M. acerina. During different cultivation periods, the differences of the colony diameters and thickness of the three fungi al reached extremely significant level. However, at the same cultivation time, the differences of the diameters and thickness among the three fungi only reached significant level. [Conclusion] The main pathogenic fungi which result in the root rot, black spot and round spot of the P. notoginseng in Wenshan are C. didymium, A. panax and M. acerina, respec-tively. When these three diseases break out at the same time, the root rot wil spread fastest, fol owed orderly by the black spot and the round spot.

Preliminary Identification of Red Pigment and Positive Correlation between the Contents of Red Pigment and Total Saponins of Panax notoginseng Fruits

[Objective] This study aimed to identify red pigment of Panax notoginseng fruits and explore the correlation between pigment content and total saponins of the fruits. [Method] The red pigment of Panax notoginseng fruits was preliminarily identi- fied with specific color reactions and UV-vis spectra, and the contents of the pigment and total saponins were determined via spectrophotometry. [Result] The red hues of the fruits were contributed by anthocyanins and/or the anthocyanidins. The contents of anthocyanins and total saponins of the fruits both decreased along with thinning of the red hues. The content difference of the anthocyanins in fruits with different red hues reached extremely significant level, but that of total saponins just reached significant level. [Conclusion] The red pigment of P. notoginseng fruits is anthocyanins which are of extremely significant positive correlation with total saponins in contents.

Responses of Photosynthetic Pigment, Phenol and Total Saponin Contents of Panax notoginseng to Supplemental UV-B under Field Conditions

[Objective] The aim of this study was to investigate the content changes and their correlations of the photosynthetic pigment,phenols,including total phenols,total flavonoids and anthocyanins,and total saponins of the one-year-old P.notoginseng plants under supplemental UV-B stress in fields.[Method] The one-year-old plants were irradiated by UV-B in field for 1 min per day,and the plants under the UV-B lamp were regarded as a circle center,achieving an annular leaf-sampling.The photosynthetic pigment,phenols and total saponins of the leaves were determined spectrophotometrically.[Result] With the increase of sampling radius,the supplemental UV-B intensity decreased significantly,the contents of chlorophyll （Chl） a,Chl b,Chl （a＋b）,carotenoid （Car） and total photosynthetic pigment （Chl＋Car） of the leaves increased extremely significantly,the Chl a/b and total phenol content （TPC） decreased extremely significantly,but the Chl （a＋b）/Car changes were not significant.The contents of total flavonoids,anthocyanins and saponins all increased due to the increasing of UV-B,displaying dose effects.The UV-B intensity was positively correlated with the Chl a/b,and negatively with the Chl a,Chl b,Chl （a＋ b）,Car and （Chl＋Car） contents; and the two of TPC,total flavonoid content （TFC）,total anthocyanin content （TAC） and total saponin content （TSC） were positively correlated,all reaching extremely significant level.The UV-B intensity was positively and significantly correlated with the total flavonoid content （TFC）,negatively and significantly with the Chl （a＋b）/Car,and positively and insignificantly with the TPC,TAC and TSC.[Conclusion] For one-year-old plants of P.notoginseng,UV-B can decrease the contents of the Chl a,Chl b,Chl （a＋b）,Car and （Chl＋Car） and increase the Chl a/b and TPC,and,furthermore,induce the increases of the TFC,TAC and TSC in a dose-dependent manner.However,UV-B can hardly change the Chl （a＋b）/Car.The supplemental UV-B of well-suited dose might be one of the effective measures to improve the TSC of P.notoginseng.

三七(Panax notoginseng)对不同光照强度的生理生态适应性研究

为探讨三七（Panax notoginseng）对不同光照强度的生理生态适应机制,本研究以二年生三七为材料,研究其在不同光照强度下的生长特性、叶片解剖结构、生物量积累与分配以及光合生理特性.结果表明：处理120d后,透光率为5.10％荫棚下三七株高、叶面积、茎粗均显著高于其他遮阴处理（P＜0.05）.随着光强降低,三七叶片厚度、栅栏组织厚度、海绵组织厚度、根生物量比、气孔导度、蒸腾速率均随之减小,而其叶生物量比、茎生物量比及水分利用速率却随之增大.透光率为46.5％ ～5.10％荫棚下,随着透光率的减小,三七净光合速率随之增大,暗呼吸速率随之减小,但在最低透光率2.80％的环境下,三七其净光合速率反而减小,暗呼吸速率反而增大.综合来看,三七表现出典型喜阴植物的特征,在光照强度相对较低的生长环境下三七的净光合速率较高,生长表现较好,根生物量积累较多.

不同颜色农膜对药用植物三七(Panax notoginseng)种苗生长的影响

通过研究不同颜色滤光农膜大棚对三七种苗形态和生物量的影响,为三七健康育苗选择适合颜色农膜提供理论依据。研究采用红、橙、黄、绿、青、蓝和紫色农膜搭建三七生长棚,在三七种苗子条膨大中期,测定与分析不同彩色农膜下三七种苗的形态特征和生物量积累。结果表明,在红色和橙色农膜大棚下三七种苗株高和冠幅最大;根数、根长及块根直径在青色农膜大棚下具有最大值;在黄色农膜大棚下三七种苗茎粗显著大于其他颜色农膜大棚下的三七种苗的茎粗;在蓝色农膜大棚下叶绿素含量相对值最高;橙色、黄色、青色和紫色农膜大棚下生物量积累最大;与其他颜色农膜相比,青色和黄色农膜下三七生物量根重比较大,绿色农膜大棚下叶重比较大。总的来看,在橙色、黄色、青色和紫色农膜大棚下三七种苗生长较好。

High-Performance Liquid Chromatographical Analysis of Ginsenosides in Panax ginseng,P.quinquef(?)lium and P.notoginseng

The compositions and contents of ginsenbsides in Panax ginseng,P.quinquefolium and P.notoginseng were determined and compared by reversed-phase High-Performance Liquid Chro- matography(HPLC).The method was performed on an Alltech Adsorbosphere HS C_(18) column,using 5×10^(-3)M NaH_2PO_4-H_3PO_4 buffer solution(pH 3.0)and acetonitrile-water(50:50)as gradient eluents. The baseline separation of ginsenosides Rb_1,Rb_2,Rb_1,Rc,Rd,Rf,Ro,and Re+Rg_1 was obtained in one analytical run.The ginsenosides are directly detected at 203 nm.The detection limit is 40μg at a signal to noise ratio of 3:1.The improved sample preparation and clean-up prior to injection with SEP-PAK C_(18)cartridge strongly reduced the front peaks caused by the impurities in the methanolic extracts of samples to afford a smooth baseline and clear background.The HPLC patterns of methanolic extracts mainly including the ginsenosides were found capable of serving as chemical fingerprints to differentiate the three species from each other.It was also found that there are no significant diffe- rences of the HPLC patterns between the wild Panax ginseng and the cultivated,the white and the red ginsengs,Chinese and Korean red ginsengs,and the tap roots of Panax ginseng collected in four consecutive months,only certain differences in contents of ginsenosides do exist.The contents of the nine major ginsenosides present in the rhizome,tap root and rootlet as well as the leaf of Panax quinquefolium were also determined and compared.

Simultaneous determination of five nucleosides and nucleobases in Panax notoginseng using high-performance liquid chromatography

Aim To quantitatively determine five nucleosides and nucleobases, including cytidine, uridine, guanosine, adenosine and uracil in different parts of Panax notoginseng. Methods Separation was performed on a Zorbax SB-Aq column using a gradient elution with mobile phase of 8 mmol^L-1 ammonium acetate aqueous solution （A） and methanol （B）. The assay was carried out at a flow rate of 1 mL·min^-1 at 25 ℃ with the diode-array detection at 260 nm. Results Cytidine, uridine, guanosine, adenosine and uracil had good linearity in the ranges of 1.79 - 57.40 μg·mL^-1 （r^2 = 1.0000）, 3.30 - 105.60 μg·mL^-1 （r^2 = 1.0000）, 3.09 - 98.80 μg·mL^ -1（r^2 = 0.9999）, 2.77 - 88.60 μg·mL^-1 （r^2 = 1.0000） and 0.38 - 12.30 μg·mL ^-1 （r^2 = 1.0000） with average recoveries of 93.9%, 96.5%, 92.7%, 93.2% and 98.8%, respectively. The content of cytidine, uridine, guanosine, adenosine and uracil in different parts of P. notogingeng were significantly different. Conclusion This is the first report on quantitative determination of nucleosides and nucleobases in P notoginseng.

Effects of total saponins of Panax notoginseng on immature neuroblasts in the adult olfactory bulb following global cerebral ischemia/reperfusion

The main active components extracted from Panax notoginseng are total saponins. They have been shown to inhibit platelet aggregation, increase cerebral blood flow, improve neurological behavior, decrease infarct volume and promote proliferation and differentiation of neural stem cells in the hippocampus and lateral ventricles. However, there is a lack of studies on whether total saponins of Panax notoginsertg have potential benefits on immature neuroblasts in the olfactory bulb following ischemia and reperfusion. This study established a rat model of global cerebral ischemia and reperfusion using four-vessel occlusion. Rats were administered total sa- ponins of Panax notoginseng at 75 mg/kg intraperitoneally 30 minutes after ischemia then once a day, for either 7 or 14 days. Total saponins of Panax notoginseng enhanced the number of dou- blecortin （DCX）＋ neural progenitor ceils and increased co-localization of DCX with neuronal nuclei and phosphorylated cAMP response element-binding/DCX＋ neural progenitor cells in the olfactory bulb at 7 and 14 days post ischemia. These findings indicate that following global brain ischemia/reperfusion, total saponins of Panax notoginseng promote differentiation of DCX＋ cells expressing immature neuroblasts in the olfactory bulb and the underlying mechanism is related to the activation of the signaling pathway of cyclic adenosine monophosphate response element binding protein.

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.

Panax notoginseng saponin attenuates hypoxia/reoxygenation-induced oxidative stress in cortical neurons

The present study monitored the effect of 2, 10, and 50 mg/L of Panax notoginseng saponin exposure following hypoxia-reoxygenation injury in fetal rat cortical neurons. Results showed that varying doses of Panax notoginseng saponin significantly enhanced the cell viability of neurons, reduced malondialdehyde content, increased superoxide dismutase activity, inhibited mRNA and protein expression of inducible and neuronal nitric oxide synthase, and decreased the release of nitric oxide in hypoxia/reoxygenation injured cells. In particular, 50 mg/L of Panax notoginseng saponin was the most effective dose. These findings suggest that Panax notoginseng saponin can attenuate neuronal oxidative stress injury caused by hypoxia/reoxygenation in a dose-dependent manner.

Panax notoginseng saponins preconditioning protects rat liver grafts from ischemia/reper- fusion injury via an antiapoptotic pathway

BACKGROUND: Ischemia/reperfusion (I/R) injury is a major cause of primary graft dysfunction and renders an al- lograft more immunogenic in orthotopic liver transplanta- tion (OLT). Panax notoginseng saponins (PNS) has been re- ported to exert protective effects against I/R injury to vari- ous organs. The objective of this study is to investigate whether PNS preconditioning protects rat liver grafts from I/R injury via an antiapoptotic pathway. METHODS: Male Sprague-Dawley rats were used as donors and recipients of orthotopic liver transplantation ( OLT) and were divided into PNS preconditioning group (group P) and normal saline control group (group N) randomly according to whether PNS (50 mg/kg) was injected intra- venously 1 hour before liver grafts harvesting, and sham group (group S). The animals were separately killed 2, 6 and 24 hours after reperfusion. Plasma samples were collect- ed for test of alanine amino-transferase (ALT) and aspartate aminotransferase (AST). Liver tissues were collected to de- tect histological changes, apoptosis and the expression of TNF-α, Bcl-2 and Caspase-3 mRNA. RESULTS: The serum levels of ALT and AST and the apop- tosis index (AI) of liver tissue in group P were lower than in group N significantly 2, 6 and 24 hours after reperfusion. Compared with group N, the expression of TNF-a and Caspase-3 mRNA was reduced significantly in group P 2 and 6 hours after reperfusion and the expression of Bcl-2 mRNA was enhanced significantly in group P 6 and 24 hours after reperfusion. CONCLUSIONS: PNS preconditioning protects liver grafts from I/R injury effectively in rat OLT via an antiapoptotic pathway. The antiapoptotic mechanisms of PNS may in- clude inhibiting the expression of TNF-a and Caspase-3 and enhancing the expression of Bcl-2.

Panax notoginseng saponins influence on transplantation of neural stem cell-derived dopaminergic neurons in a rat model of Parkinson’s disease

BACKGROUND： Dopaminergic neurons differentiated from neural stem cells have been successfully used in the treatment of rat models of Parkinson＇s disease; however, the survival rate of transplanted cells has been low. Most cells die by apoptosis as a result of overloaded intracellular calcium and the formation of oxygen free radicals. OBJECTIVE： To observe whether survival of transplanted cells, transplantation efficacy, and dopaminergic differentiation from neural stem cells is altered by Panax notoginseng saponins （PNS） in a rat model of Parkinson＇s disease. DESIGN, TIME AND SETTING： Cellular and molecular biology experiments with randomized group design. The experiment was performed at the Animal Experimental Center, First Hospital of Sun Yat-sen University from April to October 2007. MATERIALS： Thirty-two adult, healthy, male Sprague Dawley rats, and four healthy Sprague Dawley rat embryos at gestational days 14-15 were selected. The right ventral mesencephalon was injected with 6-hydroxydopamine to establish a model of Parkinson＇s disease. 6-hydroxydopamine and apomorphine were purchased from Sigma, USA. METHODS： Neural stem cells derived from the mesencephalon of embryonic rats were cultivated and passaged in serum-free culture medium. Lesioned animals were randomly divided into four groups （n = 8）： dopaminergic neuron, dopaminergic neuron ＋ PNS, PNS, and control. The dopaminergic neuron group was injected with 3 μL cell suspension containing dopaminergic neurons differentiated from neural stem cells. The dopaminergic neurons ＋ PNS group received 3 μ L dopaminergic cell suspension combined with PNS （250 mg/L）. The PNS group received 3 μL PNS （250 mg/L）, and the control group received 3 μL DMEM/F12 culture medium. MAIN OUTCOME MEASURES： The rats were transcardially perfused with 4% paraformaldehyde at 60 days post-grafting for immunohistochemistry. The rats were intraperitoneally injected with apomorphine （0.5 mg/kg） to induce rotational behavior. RESULTS： Cell counts of tyrosine hydroxylase-positive neurons in the dopaminergic neuron ＋ PNS group were （732±82.6） cells/400-fold field. This was significantly greater than the dopaminergic neuron group [（326 ± 34.8） cells/400-fold field, P 〈 0.01]. Compared to the control group, the rotational asymmetry of rats that received dopaminergic neuron transplants was significantly decreased, beginning at 20 days after operation （P 〈 0.01）. Rotational asymmetry was further reduced between 10-60 days post-surgery in the dopaminergic neuron ＋ PNS group, compared to the dopaminergic neuron group （P 〈 0.01）. CONCLUSION： Panax notoginseng saponins can increase survival and effectiveness of dopaminergic neurons differentiated from neural stem cells for transplantation in a rat model of Parkinson＇s disease.