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.

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.

Ginsenoside-Rg_6, a Novel Triterpenoid Saponin from the Stem-Leaves of Panax ginseng C. A. Mey.

A novel dammarane-type triterpene oligoglycoside, named ginsenoside-Rg6 3, was isolated from the stem-leaves of Panax ginseng C. A. Mey., together with two known ones, 20(S)-ginsenoside-Rg2 1 and 20(R)-ginsenoside-Rg2 2. On the basis of chemical and physicochemical evidence , the structure of ginsenoside-Rg6 have been elucidated as 6-O-(-L-rhamnosyl-(1?2)-(-D-glucopyranosyl-dammarane-(E)-20(22), 24-diene-3(, 6(, 12(-triol.

Determination of Saponins in Leaf of Panax Ginseng C. A. Mey. by High Performance Liquid Chromatography

A high performance liquid chromatography（HPLC） with UV detection was established for simultaneous determination of saponins in the leaf of Panax ginseng C. A. Mey. Nine ginsenosides（Rbl, Rb2, Rb3, Rc, Rd, F1, F2, F3, F5） and notoginsenoside Fe（NFe） were studied. Among the saponins, the ginsenosides F1, F2, F3, F5 and NFe were determined by HPLC-UV method for the first time. The determination of the ginsenosides via the HPLC-UV method was performed on a reversed-phase C18 column with gradient elution in 40 min. The linearity, precision, accuracy, and detection limit for determining the saponins were studied and the samples from different areas in China were analyzed. The HPLC-ESI-MS was used to identify the saponins. The results indicate that the HPLC-UV provided a good accuracy, reproducibility and sensitivity for the determination of the ten saponins.

Structure Analysis of Pectin SB_(1-1) from the Root of Panax ginseng

A water-soluble pectin SB_~1-1 was isolated and purified from the root of Panax ginseng C. A. Mey. The HPLC analysis indicates that SB_~1-1 is homogenous. Its molecular weight was estimated via gel filtration to be 10000. The GC analysis indicated that it contains the monosaccharides of GalA, Gal, Ara and Rha. Their molar ratio is 2.10∶1.00∶0.12∶0.13. Partial hydrolysis with acid, pectinase treatment, periodate oxidation, Smith degradation, methylation analyses, GC/MS analyses and NMR analyses were used for the structure analyses of SB_~1-1 . The results reveal that SB_~1-1 has a lower branched structure. The main chain is composed of GalA and Gal; the inner part is α-1,4-linked-GalA; the border is 1,4-linked-Gal. Some of the 1,4-linked-GalA and 1,4-linked-Gal residues are substituted at O6. On an average, there is one branch for every ten hexose residues. The side chain is composed of 1,6-linked-Gal and 1,3,6-linked-Gal. The nonreduced end is composed of Rha, Ara and Gal. The main glycosidic link of SB_~1-1 has an α configuration.

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.

Active compounds in RenShenJian decoction ameliorate insulin resistance in vitro

Background:RenShenJian decoction,a combination of Pueraria lobata(Willd.)Ohwi and Panax ginseng C.A.Mey,has been used in China since the Song Dynasty(960-1279 C.E.)to relieve symptoms of diabetes mellitus.However,the key compounds in RenShenJian that ameliorate insulin resistance remain unclear.This study identified the anti-diabetic compounds in RenShenJian by rescuing the decreased function of adenosine 5’-monophosphate-activated protein kinase(AMPK),sirtuin 3(SIRT3),or glucose transporter isoform 4(GLUT4).Methods:After streptozotocin-induced diabetic mice were treated with RenShenJian,fasting blood glucose levels and protein expression of SIRT3,p-AMPK,and AMPK were determined.Compounds from RenShenJian in plasma were monitored using multiple responses by liquid chromatography-mass spectrometry.Additionally,two insulin-resistant cell models were incubated with compounds identified in RenShenJian in the blood.Glucose uptake was determined using the fluorescent analog 2-(N-(7-nitrobenz-2-oxa-1,3-dia-xol-4-yl)amino)-2-deoxyglucose.Protein expression levels of p-AMPK,AMPK,SIRT3,and GLUT4 were detected by western blotting.Results:RenShenJian decreased FBG levels and upregulated SIRT3 expression and AMPK phosphorylation in diabetic mice.Thirteen RenShenJian extracts were identified in the blood,11 of which increased the ratios of 2-(N-(7-nitrobenz-2-oxa-1,3-dia-xol 4-yl)amino)-2-deoxyglucose uptake in two insulin-resistant cell models.Nine extracts increased AMPK phosphorylation,nine increased SIRT3 expression,and six elevated GLUT4 expression in palmitate-induced HepG2 cells.Five extracts-puerarin,puerarin 6″-O-xyloside,genistein,ginsenoside Rb1,and ginsenoside Rd-simultaneously activated AMPK,SIRT3 and GLUT4.Conclusion:A series of compounds in RenShenJian that target AMPK,SIRT3,and/or GLUT4 was confirmed and indicate the chemical material basis of amelioration of insulin resistance by RenShenJian.

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.

Research progress on chemical diversity of saponins in Panax ginseng

Saponins,the major bioactive components of Panax ginseng C.A.Mey.,are gradually emerging as research hotspots owing to the possession of various pharmacological activities.This review updates the ginsenosides list from P.ginseng and the steam-processed ginseng(red ginseng and black ginseng)up to 271 by June of 2024,encompassing 243 saponins from different parts of P.ginseng(roots,stems,leaves,flowers,berries,and seeds),103 from red ginseng,and 65 from black ginseng,respectively.Among 271 saponins,there are a total of 249(1–249)dammarane type(with a–z subtypes)tetracyclic triterpene saponins reported from each part of P.ginseng and steam-processed ginseng,two(250–251)lanostane type tetracyclic triterpene saponins identified from red ginseng,18(252–269)oleanane type pentacyclic triterpenoid saponins discovered from each part of P.ginseng and steam-processed ginseng,and two(270–271)ursane type pentacyclic triterpenoid saponins reported from red ginseng.Overall,this review expounds on the chemical diversity of ginsenosides in various aspects,such as chemical structure,spatial distribution and subtype comparison,processed products,and transformation.This facilitates more indepth research on ginsenosides and contributes to the future development of ginseng.

HPLC法测定人参、西洋参和三七不同部位中人参皂苷的含量

目的:为了探讨人参、西洋参和三七中人参皂苷的资源含量,以确保人参、西洋参和三七中人参皂苷资源充分被利用,为开发以人参皂苷为主的创新药物提供科学数据。方法:采用高效液相色谱法对人参、西洋参和三七不同部位中人参皂苷的含量进行测定。色谱条件为:Agilent 1100 Series 高效液相色谱仪;色谱柱为德国 Nucleosil-C_(18)(4.6 mm×150 mm,5μm);流动相为水-乙腈梯度洗脱,流速1.5 mL·min^(-1);A为水,B为乙腈;梯度洗脱程序为:0～16 min,56%B;16～20 min,56%→100%B;20～38 min,100%B;38～45 min,100%→56%B;45～60 min,56%B;60～70 min,56%B。所有组分均70 min 内出完。检测波长203 nm,柱温35℃,灵敏度为0.02AUFS。线性关系考察r=0.9994;精密度试验 RSD=0.26%,平均回收率为99.88%,重现性试验 RSD=2.0%,分离度为R=3.042。结果:人参须根含有较高的人参皂苷 Rb_1(1.082%),人参茎叶则含有较高的人参皂苷 Rc(1.002%)、Re(3.430%)和 Rg_1(1.303%);西洋参根含有较高的人参皂苷 Rb_1(2.213%)和 Re(0.9188%),西洋参芦头中含有较高的人参皂苷 Rb_1(2.840%)和Re(1.224%);三七根含有较高的人参皂苷 Rb_1(2.163%)和 Rg_1(2.633%),三七芦头中含有较高的人参皂苷 Rb_1(4.376%)和 Rg_1(4.145%)。结论:高效液相色谱法分离、分析人参皂苷效果好、准确、迅速、简便,也可作为评价人参属植物质量的有效分析方法。建议对人参、西洋参和三七中含量较高的人参皂苷进行提取分离,直接用于创新药物的开发。

人参RAPD产物的限制性内切酶消化

为了鉴定随机扩增产物的同源性以及更有效地利用DNA分子标记研究人参种质资源，我们首次将毛细管PCR扩增的RAPD反应产物进行限制性内切酶消化并有效的进行了酶切位点分析，结果证明PCR-RFLP是筛选人参特异DNA分子标记的一种简单易行的新方法，可为药用植物种质资源研究提供另一有用的工具。

基于GC-MS法的不同品种人参中脂肪酸成分及含量分析

[目的]研究不同品种人参的脂肪酸成分及相对含量,为人参的良种选育工作提供一项重要指标。[方法]采用索式提取法提取,GC-MS法测定相对含量。[结果]鉴定出20种脂肪酸,其中主要为(Z,Z)-9,12-十八碳二烯酸、棕榈酸和(Z,Z,Z)-9,12,15-十八碳三烯酸;不同品种人参脂肪酸单体的含量存在差异,主要为①(Z,Z)-9,12-十八碳二烯酸含量大马牙47.896%、二马牙50.901%、圆膀圆芦50.254%、长脖49.456%,其中二马牙含量最高;②棕榈酸含量大马牙12.370%、二马牙12.535%、圆膀圆芦12.614%、长脖11.983%,圆膀圆芦含量最高;③(Z,Z,Z)-9,12,15-十八碳三烯酸含量大马牙10.407%、二马牙12.487%、圆膀圆芦11.936%、长脖12.026%,二马牙含量最高。[结论]用该法可鉴别出不同品种的人参,有望成为人参品种选育的一项重要手段。

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、西洋参的基因组DNA的RAPD分析

用Operon公司的E．F．C组引物对北京和吉林左家特区的栽培人参和西洋参四大居群各10个样本进行RAPD分析，筛选出18个引物，可以检测到102个多态性位点，其中67．6％表现出多态性。在北京栽培人参、西洋参。吉林左家特区栽培人参、西洋参中分别检测到74，86，74，9O个标记位点，在各居群中均有丰富的多样性存在。西洋参的遗传多态性明显高于人参。聚类分析结果表明，同种不同居群的样本首先聚在一起，种间遗传分化明显大于地理环境造成的分化。由OPF02，OPF04，OPF07，OPF20，OPG12，OPG13，OPG14扩增的指纹图谱中可以找到不同种，不同居群的标记带型及居群内部的丰富遗传多样性。

人参毛状根及愈伤组织中ATP动态变化的生物发光法测定

ATP content of cultured callus and hairy root introducted by Ri plasmid of \%Panax ginseng \%C. A. Mey was detected by bioluminescence. The result showed that ATP content in hairy root is obviously higher than that in callus within a growth cycle. The highest ATP content in hairy root is 26.6×10\+\{-12\}mmol/g (fresh weight), while ATP in callus is 2.68×10\+\{-12\}mmol/g (fresh weight). Total saponin content in hairy root reaches 2.486% (dry weight), while the saponin in callus is 1.105% (dry weight). The hairy root inducted by Ri plasmid has vigorous ability in secondary metabolism for ginseng saponin synthesis.

人参根际土壤微生物PCR-DGGE反应体系优化

[目的]优选人参根际土壤微生物的PCR-DGGE反应体系。[方法]以种植1年的人参土壤为材料,利用PCR-DGGE技术,对反应体系中的几种重要参数不同梯度进行了优化研究,包括模板、dNTPs、引物及Mg2+的用量。[结果]最适宜的反应体系为:模板DNA浓度为0.8μg/μl,dNTPs浓度为0.2 mmol/L,引物浓度为0.4μmol/L,Mg2+浓度为1.5 mmol/L。[结论]该方法简便快捷,为进一步研究人参根际土壤微生物多样性奠定了基础。

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.

Metabolomics analysis reveals the renal protective effect of Panax ginseng C. A. Mey in type 1 diabetic rats

The dry root and rhizome of Panax ginseng C. A. Mey has garnered much interest owing to its medicinal properties against diabetes and cardiovascular diseases. In this study, an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS)-based metabolomics approach was used to illustrate the therapeutic mechanisms of ginseng extract on the serum and urinary metabolic profiles in streptozotocin-induced type 1 diabetes mellitus (T1DM) rats. Pharmacological and renal parameters in response to the administration of ginseng were also evaluated. In total, 16 serum endogenous metabolites and 14 urine endogenous metabolites, including pyruvic acid, indoleacetic acid, and phenylacetylglycine, were identified as potential biomarkers for diabetes. Pathway enrichment and network analysis revealed that the biomarkers modulated by ginseng were primarily involved in phenylalanine and pyruvate metabolism, as well as in arginine biosynthesis. Moreover, the levels of several renal injury-related biomarkers in T1DM rats were significantly restored following treatment with ginseng. The administration of the extract helped maintain tissue structure integrity and ameliorated renal injury. The findings suggest that the regulatory effect of ginseng extract on T1DM involves metabolic management of diabetic rats, which subsequently attenuates T1DM-induced early renal dysfunction.

Identification of anti-inflammatory components in Panax ginseng of Sijunzi Decoction based on spectrum-effect relationship

Objective: This study aimed to identify the main medicinal active components of Panax ginseng(P. ginseng) in the compatibility environment of clinical application. For this purpose, the anti-inflammatory ingredients of P. ginseng were investigated based on its therapeutic effect in Sijunzi Decoction(SJD) which is a widely used traditional Chinese formula.Methods: The fingerprints of 10 batches of SJD consisting of different sources of P. ginseng were established by UPLC technique to investigate the chemical components. At the same time, the antiinflammatory effects of these components were evaluated by dextran sulfate sodium-induced ulcerative colitis mouse model. Grey relational analysis was applied to explore the correlation degree between fingerprints and anti-inflammatory effects in SJD. Lipopolysaccharide-stimulated RAW264.7 murine macrophages were established to evaluate the anti-inflammatory action of the screened effective substances of P. ginseng.Results: According to grey relational analysis, notoginsenoside R1, ginsenoside Rg2 and ginsenoside Rb3of P. ginseng were the major anti-inflammatory contributions in SJD. They had been proven to be closely associated with the anti-inflammatory process of SJD and displayed a close effect compared with SJD by LPS-stimulated RAW264.7 murine macrophages.Conclusion: Our work provides a general strategy for exploring the pharmacological ingredients of P. ginseng in traditional Chinese formulas which is beneficial for establishing the quality standards of traditional herbs in traditional Chinese medicine prescription based on their clinical therapeutic effect.