A new triterpene saponin from Panax japonicus C.A.Meyer var major(Burk.) C.Y.Wu et K.M.Feng

One new Iriterpene saponin was isolated from Panaxjaponicus C. A. Meyer var major （Burk.） C. Y. Wu et K. M. Feng, and established as oleanolic acid 3-O-[β-D-glucopyranosyl-（1 →2）-β-D-glucuronopyranosyl-6＇-O-n-butyl ester] which showed mod- erate antitumor activities against the A2780 cells and OVCAR-3 cells. Its structure was established by means of spectral data, particularly NMR, including HSQC and HMBC techniques.

Research Advances on Panax japonicus and its Approximation Varieties in Tujia Nationality

Panax japonicus and its approximation varieties,such as Rhizoma Panacis Majoris and Panax japonicus C. A. Mey. var.major （Burk.） C.Y. Wu et K.M. Feng belong to Panax,which are less commonly used traditional Chinese medicine. Because of similar traits and effectiveness,they were always used as one type of medicine for a long time. Aiming at this phenomenon,the chemical composition and contents of P. japonicus and its approximation varieties from different area were compared in order to provide a chemical basis for clarifying the classification of the genus.

Optimizing SSR-PCR system of Panax ginseng by orthogonal design

An orthogonal design was used to optimize SSR-PCR amplification system using Panax ginseng genomic DNA as template. Four levels of five factors （DNA template, Taq DNA polymerase, Mg^2＋, primer, and dNTP） and annealing temperature have been tested separately in this system. The results demonstrated the reaction efficiency was affected by these factors. Based on the results, a stable, productive and reproducible PCR system and cycling program for amplifying a ginseng SSR locus were obtained： 20 μL system containing 1.0 U Taq DNA polymerase, 2.0 mmol·L^-1 Mg^2＋, 0.2 mmol·L^-1 dNTPs, 0.3 μmol·L^-1 SSR primer, 60 ng· μla^-1 DNA template, performed with a program of 94℃ for 5 min, 94℃ for 30 s, annealing at 56.3℃ for 30 s, 72℃ for 1 min, 37 cycles, finishing at 72℃ for 7 min, and storing at 4℃.

Antagonistic Effects of Sphingomonas and Pseudomonas aeruginosa on 4 Kinds of Pathogenic Bacteria of Ginseng

[Objectives]To explore effective biocontrol methods for diseases in the process of ginseng cultivation,and develop an efficient and environmentally friendly biocontrol agent.[Methods]In this study,2 strains were isolated from biogas slurry,and Cylindrocarpon destructans(XF),Fusarium solani(GF),Botrytis cinerea Pers(HM)and Alternaria panax Whetz(HB)were used as test materials.The strains were isolated and identified by dilution plate method,16S rDNA sequence identification method,confrontation culture method,filter paper method and ultraviolet spectrophotometer method,and the bacteriostatic activity and bacteriostatic rate were tested.[Results]Strain 15(Sphingomonas)and strain 19(Pseudomonas aeruginosa)were screened out through identification and analysis,and they grew stably within 8-10 d.The bacteriostatic rates of strain 15 against A.panax and B.cinerea were 47.37%and 43.40%,respectively,and the bacteriostatic rates of strain 19 against A.panax and B.cinerea were 62.30%and 63.27%,respectively.The bacteriostatic activity of the extract of strain 19 increased with the increase of OD_(600) value,and the bacteriostatic effect was optimal when the OD_(600) value was in the range of 0.8-1.0,up to 70%,so it had a strong biocontrol potential.[Conclusions]This experiment provides convenience for more effective inoculation,establishes a fast,simple and accurate method for the determination of the best bacteriostatic rate of P.aeruginosa culture solution to HM,and lays a foundation for large-scale culture of P.aeruginosa culture solution.Besides,it is expected to provide a theoretical basis for the efficient control of ginseng B.cinerea in field production,use it for the prevention and control of ginseng shoot diseases,and provide a reference for the efficient and diverse development of biocontrol agents for ginseng shoot diseases.

Triterpenoid Saponins from Tu Jia Ethnomedicine Bai San Qi and Their Cytotoxicity on Hep G2 and BGC-823 Cell Lines

Objective Tu Jia ethnomedicine is a unique medical system inherited and adhibited by Tu Jia minority living in central-south China. Panax japonicus C. A. Mey.(Bai San Qi,白三七) is recognized as an effective and rare medicinal plant to treat weakness, fatigue and rheumatism in Tu Jia ethnomedicine. This paper is to discover more substance evidence for the application of Tu Jia ethnomedicine. Methods Column chromatography and preparative high performance liquid chromatography (HPLC) was applied for isolation and purification;1H-NMR, 13C-NMR, 1H-1H COSY, HSQC and HMBC NMR spectra were applied for structure identification;Methyl thiazolyl tetrazolim (MTT) assays were applied for cytotoxicity evaluation. Results Totally 12 known compounds were isolated by column chromatography and preparative HPLC from rhizomes of Panax japonicus C. A. Mey.(Bai San Qi,白三七). Structures of these compounds were identified by their NMR spectra. All the 12 compounds were triterpenoid saponins. Five of them were oleanolic acid type, and the remaining 7 were dammarane type. Eleven compounds were assayed for their cytotoxic activity against Hep G2 human liver cancer cell lines and BGC-823 human gastric cancer cell lines. Three of the 11 showed relatively dominant cytotoxicity against these cell lines. Conclusions A total of 12 known compounds have been identified from Panax japonicus C. A. Mey.(Bai San Qi,白三七);NMR spectra of compounds with similar skeletons showed regular characteristics;3 compounds showed relatively dominant cytotoxicity against Hep G2 and BGC-823 cancer cell lines, and the result can be valued as weak while setting the taxol as a positive control.

Preparation and antiviral effects of polysaccharides from Panax japonicus

Water-soluble polysaccharides were prepared from Panax japonicus by hot water extraction and ethanol precipitation.The polysaccharides were further purified by ion exchange chromatography to obtain neutral and acidic polysaccharides.The neutral polysaccharide fraction mainly contained Glc(90.2%),which was a glucan fraction.The acidic polysaccharide fraction mainly contained GalA(43.6%),Gal(21.7%),and Ara(15.4%),with a degree of methyl-esterification of 20.3%,which was a pectic polysaccharide.The acidic polysaccharide of Panax japonicus could effectively inhibit the replication of human seasonal influenza virus H1N1 and canine influenza virus H3N2 in MDCK cells and A549 cells and significantly reduce the virus titer in infected cells.It also effectively inhibited the number of infected cells of the SARS-CoV-2 South Africa strain and the Omicron strain.The acid polysaccharide of Panax japonicus showed good efficacy against influenza virus and COVID-19 infection,which could be used as a potential antiviral candidate drug molecule in the future.

超临界CO_2反相微乳萃取人参中人参皂苷的研究

目的提高超临界CO2萃取人参中人参皂苷的萃取率。方法采用二-(2-乙基己基)磺基琥珀酸钠(AOT)/乙醇/水/超临界CO2反相微乳对人参皂苷的萃取进行研究。结果在萃取压力25MPa、温度45℃、时间4h、CO2流量为2L/h条件下,超临界CO2反相微乳萃取的人参皂苷萃取率是乙醇/水/超临界CO2萃取的3.2倍。人参皂苷的萃取率随加入的水量、萃取压力的增大而增大,随AOT浓度、萃取温度的升高先增大后减小。萃取人参皂苷时,采用适量水先浸泡物料与萃取过程中加入水相比,人参皂苷的萃取率要提高30%。结论结合实验结果与理论探讨,超临界CO2反相微乳萃取人参皂苷的机制主要是其形成的极性水池增大了对人参皂苷的溶解度。

酶法修饰人参茎叶总皂苷及其HPLC图谱研究

目的研究生物酶制剂对人参茎叶总皂苷的修饰及其修饰前后HPLC图谱的变化。方法利用复酶制剂A对人参茎叶总皂苷进行酶法修饰,采用TLC法鉴别修饰前后的变化,并比较修饰前后的HPLC图谱的变化。结果在设定的积分事件下,复酶制剂A修饰人参茎叶总皂苷后色谱特征峰由34个减至16个,其中有13个特征色谱峰相对峰面积和相对峰高均明显降低,21个色谱峰检测不到相应信号,产生4个明显增加或新色谱峰。结论复酶制剂A能够有效地修饰人参茎叶总皂苷,采用HPLC图谱能够很好的监测其变化。

珠子参ISSR-PCR反应体系的建立及优化

目的：建立珠子参的ISSR-PCR适宜扩增反应体系和程序，为其深入研究奠定基础。方法采用新型植物基因组DNA提取试剂盒提取珠子参总DNA ，并用正交设计实验对其ISSR反应体系条件进行筛选及优化。结果提取的基因组DNA纯度和完整性较好，A260/A280值在1．8～2．0之间，DNA无降解现象，可满足ISSR-PCR扩增要求。珠子参ISSR分析的最适反应体系为：在20μL PCR反应体积中，含10 ng模板DNA、0．20 mmol·L -1 dNTPs、1．5μmol·L -1引物、3U TaqDNA 聚合酶、2μL 10× PCR Buffer、3 mmol·L -1 Mg2＋。扩增程序为：95℃预变性5 min ，94℃变性30s，58℃退火30 min ，72℃延伸1 min ，循环40次，72℃延伸10 min。由此可获得带型丰富和清晰可辨的DNA指纹图谱。结论建立的ISSR-PCR反应体系可用于研究珠子参的遗传变异和遗传多样性。

人参总皂苷体外定向诱导CD34^+细胞分化为粒系血细胞的研究

目的研究人参总皂苷(TSPG)协同造血生长因子对CD34+细胞体外定向诱导分化为粒系血细胞的影响。方法应用阴性磁性分选策略,以StemsepTM系统从正常人骨髓细胞中分离CD34+造血干/祖细胞(HSC/HPC),通过甲基纤维素半固体培养法检测TSPG诱导CD34+HSC/HPC向粒系祖细胞集落(CFU-GM)增殖与分化的能力;在SCF+IL-3+GM-CSF+G-CSF(SIGG)液体培养体系中加入不同剂量的TSPG,检测对CD34+HSC/HPC增殖形成CFU-GM的影响以及CD33+细胞比例的变化。结果TSPG(10～50μg/mL)均能提高CD34+细胞形成CFU-GM的集落产率,以TSPG20μg/mL效果最为明显;不同质量浓度的TSPG协同造血生长因子在液体培养体系中诱导CD34+细胞2周,观察粒系血细胞的分化,TSPG10～70μg/mL均可不同程度地提高细胞总数、CFU-GM扩增倍数及CD33+细胞比例,TSPG20μg/mL是液体培养诱导CD34+细胞向粒系分化的最佳质量浓度。结论TSPG协同造血生长因子能促进CD34+细胞定向诱导分化为粒系血细胞。

GC-MS法检测湘产扣子七、白三七挥发性成分含量研究

目的:比较扣子七、白三七中挥发油的化学成分含量。方法:分别采用水蒸气蒸馏法提取扣子七、白三七的挥发性成分,均采用GC-MS联用仪测定提取物,以峰面积归一化法测定各成分的相对含量。结果:从扣子七提取物中鉴定39种组分,占总峰面积的96.35%;从白三七提取物中鉴定41种组分,占总峰面积的97.17%。结论:两种药材共有成分在5%以上的有4种,分别为斯巴醇、棕榈酸、棕榈酸乙酯和亚油酸乙酯,该方法为扣子七和白三七的综合开发提供了参考依据。

Panax japonicus and chikusetsusaponins: A review of diverse biological activities and pharmacology mechanism

Panax japonicus, which in the Tujia dialect is known as "Baisan Qi" and "Zhujieshen", is a classic "qi" drug of Tujia ethnomedicine and it has unique effects on disease caused by "qi" stagnation and blood stasis.This paper serves as the basis of further scientific research and development of Panax japonicus. The pharmacology effects of molecular pharmacology were discussed and summarized. P. japonicus plays an important role on several diseases, such as rheumatic arthritis, cancer, cardiovascular agents, and this review provides new insights into P. japonicus as promising agents to substitute ginseng and notoginseng.

Spatio-Temporal Expression Pattern of Six Novel Candidate Genes in Ginsenoside Biosynthesis from Panax ginseng C. A. Meyer

Abstract: To explore the mode of the spatio-temporal expression of six newly discovered ginsenoside biosynthesis candidate gene transcripts, both Northern blotting and semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) were used to elucidate the mRNA expression levels of the transcripts in various tissues and organs of Panax ginseng C. A. Meyer during different growth development stages. The six gene transcripts were all differentially expressed in cultured callus, root, stem, leaf, and seed. The mRNA expression levels were significantly higher in four-year-old roots than in one-year-old roots, and results of semi-quantitative RT-PCR assays were in accordance with those of Northern blotting analyses. The results strongly suggest that all six genes were differentially expressed at root-specific developmental stages. In particular, when a quiescent early stage culture suspension of P. ginseng cells was exposed to the ginsenoside biosynthesis-promoting elicitor Aspergillus niger polysaccharide, the GBR6 gene transcript response showed time-dependent increments and was parallel with ginsenoside productivity (P < 0.01). Overexpressionof the GBR6 gene is likely to play a critically important role in the biosynthesis of ginsenosides. The results of the present study provided a background for the further elucidation of the structure and physiological function of these six candidate genes.

Penicillium sp.YJM-2013 induces ginsenosides biosynthesis in Panax ginseng adventitious roots by inducing plant resistance responses

Objective:Fusarium oxysporum is a common pathogenic fungus in ginseng cultivation.Both pathogens and antagonistic fungi have been reported to induce plant resistance responses,thereby promoting the accumulation of secondary metabolites.The purpose of this experiment is to compare the advantages of one of the two fungi,in order to screen out more effective elicitors.The mechanism of fungal elicitor-induced plant resistance response is supplemented.Methods:A gradient dilution and the dural culture were carried out to screen strains.The test strain was identified by morphology and 18 s rDNA.The effect of different concentrations(0,50,100,200,400 mg/L)ofPenicillium sp.YJM-2013 and F.oxysporum on fresh weight and ginsenosides accumulation were tested.Signal molecules transduction,expression of transcription factors and functional genes were investigated to study the induction mechanism of fungal elicitors.Results:Antagonistic fungi ofF.oxysporum was identified as Penicillium sp.YJM-2013,which reduced root biomass.The total ginsenosides content of Panax ginseng adventitious roots reached the maximum(48.95±0.97 mg/g)treated with Penicillium sp.YJM-2013 at 200 mg/L,higher than control by 2.59-fold,in which protopanoxadiol-type ginsenosides(PPD)were increased by 4.57 times.Moreover,Penicillium sp.YJM-2013 activated defense signaling molecules,up-regulated the expression of PgWRKY 1,2,3,5,7,9 and functional genes in ginsenosides synthesis.Conclusion:Compared with the pathogenic fungi F.oxysporum,antagonistic fungi Penicillium sp.YJM-2013 was more conducive to the accumulation of ginsenosides in P.ginseng adventitious roots.Penicillium sp.YJM-2013 promoted the accumulation of ginsenosides by intensifying the generation of signal molecules,activating the expression of transcription factors and functional genes.

植物激素对人参毛状根生长和皂甙含量的影响

就植物激素IAA、IBA、NAA、2,4 D和6 BA对人参(PanaxginsengC.A.Meyer)毛状根生长及皂甙含量的影响进行了研究。结果表明,4种生长素在适宜的浓度下均可不同程度地促进人参毛状根的生长以及皂甙的积累,同时能影响单体皂甙的分布。NAA和IBA能显著促进毛状根的生长,其中0.500mg LIBA能显著促进毛状根生长和总皂甙的积累。细胞分裂素6 BA在较低浓度时虽然对生长无明显的促进作用,但对皂甙积累有利,同时显著促进单体皂甙Rb1的积累,增大Rb1在总甙中所占的比例。

人参农家类型遗传多样性的ISSR分析

目的探讨人参农家类型的遗传多样性和亲缘关系,为人参的栽培和选育种提供遗传学依据。方法采用ISSR分子标记分析5种人参农家类型7个居群120个样本的遗传多样性。结果人参农家类型有较丰富的遗传多样性,平均多态位点百分率为48.85%;不同农家类型的遗传多样性水平有差异,和其他农家类型相比,长脖和竹节芦的遗传变异较小,且两者之间的相似性系数高达97%;不同产地的同一人参农家类型间也存在很大的遗传差异。结论研究表明人参农家类型的遗传差异主要存在于各类型内部,而且可能更多地存在于同一类型的不同居群内部。为促进人参新品种的选育,有必要在现有的栽培群体中补充不同产地的同一类型的种质资源。

竹节参总皂苷预处理对冠脉结扎致大鼠急性心肌缺血损伤的影响

目的:研究竹节参总皂苷预处理对冠脉结扎致大鼠急性心肌缺血损伤的影响,并初步探讨其可能的机制。方法:实验大鼠随机分为假手术组、模型组和竹节参总皂苷组;治疗组大鼠给予竹节参总皂苷,预防性给药7 d后,行结扎大鼠冠状动脉术制作急性心肌缺血损伤模型,术后12 h进行大鼠心电图和血流动力学检测,然后取血进行血液乳酸脱氢酶(LDH)、肌酸激酶(CK)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量分析;取心脏,分别进行心肌组织2,3,5-氯化三苯基四氮唑染色和HE染色。结果:竹节参总皂苷能明显改善心功能,对抗冠脉结扎诱导的心电图T波降低,显著降低心肌梗死面积,减少心肌缺血性损伤;降低血清中CK、LDH活性和MDA含量,增加血清中SOD、CAT活性。结论:竹节参总皂苷对冠脉结扎致急性心肌缺血损伤具有较好的保护作用,抗氧化损伤可能是其作用机制之一。

微生物对人参果总皂苷中人参皂苷化合物K的转化作用

目的利用微生物转化法对人参果总皂苷(SFPG)进行生物转化,制备人参皂苷化合物K(C-K)。方法从种植人参的土壤中筛选、分离4种微生物m14、m3、m8、m9对SFPG进行转化,筛选最佳活性菌株;以C-K的量为指标,采用TLC和HPLC法检测微生物的转化结果。结果真菌m14为转化C-K的最佳活性菌株,最佳转化条件为转化时间6d,转化温度30℃,摇床转数160r/min,转化初始pH值为自然pH5.5,转化底物质量浓度为120mg/mL。转化后的C-K的量是转化前的41.65倍。结论真菌m14转化SFPG中C-K的专属性强、转化效率高,为C-K的工业化生产提供了一条新的途径。

正交试验优化超高压提取人参中人参皂苷的工艺研究

目的 研究超高压提取人参中人参皂苷的最佳工艺。方法 采用超高压技术常温提取,正交试验优化,分光光度法检测人参总皂苷的含量。结果 超高压提取人参皂苷的最佳提取工艺参数为:提取溶剂为5 0 %乙醇,固液比为1∶75 ,提取压力为5 0 0 MPa,提取时间2 m in,人参皂苷的得率高达7.76 %。结论 超高压提取工艺具有提取效率高、时间短、能耗低、杂质含量少等优点。

响应面法优化人参不定根中总皂苷的提取工艺

以人参不定根为原材料,利用乙醇提取人参总皂苷,选择提取温度、乙醇浓度和料液比为主要影响因素,以人参总皂苷得率作为响应值,利用响应面法优化人参不定根总皂苷的提取工艺。根据Box-Behnken实验设计原理,采用三因素三水平的响应面分析法,建立了人参不定根总皂苷得率与三个因素变化的二次回归方程。依据回归模型进行计算机模拟及绘制曲面图,了解人参不定根总皂苷得率随主要因素水平的变化趋势及优化点。通过对各因素显著性和交互作用的分析,得到最佳工艺条件为:提取温度70.00℃、乙醇浓度73.50%、料液比1∶34.50(g/m L),此时人参皂苷的得率为1.86±0.01%。