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.

人参基因组DNA的提取及RAPD-PCR反应条件的优化

探讨了人参基因组DNA提取方法及RAPD-PCR反应条件的优化.结果表明,用改良的CTAB方法提取的DNA均达到了RAPD-PCR分析的要求;优化的RAPD-PCR反应条件为:在25μL RAPD-PCR反应体系中,模板DNA 20 ng,dNTP 200μmol/L,MgCl21.5mmol/L,10×Buffer 2.5μL,引物浓度0.4μmol/L,Taq DNA聚合酶1.0 U,其余以ddH2O定容至25μL.PCR反应程序为:94℃预变性5 min;94℃45 s,40℃退火1 min,72℃1 min,共40个循环;最后在72℃延伸5 min.

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.

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。

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。

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。

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.

Identification of microRNA and analysis of target genes in Panax ginseng

Objective: Ginsenosides, polysaccharides and phenols, the main active ingredients in Panax ginseng, are not different significantly in content between 3 and 5 years old of ginsengs called Yuan ginseng and more than ten years old ones called Shizhu ginseng. The responsible chemical compounds cannot fully explain difference in efficacy between them. According to reports in Lonicerae Japonicae Flos(Jinyinhua in Chinese) and Glycyrrhizae Radix et Rhizoma(Gancao in Chinese), microRNA may play a role in efficacy,so we identified microRNAs in P. ginseng at the different growth years and analyzed their target genes.Methods: Using high-throughput sequencing, the RNA-Seq, small RNA-Seq and degradome databases of P. ginseng were constructed. The differentially expressed microRNAs was identified by qRT-PCR.Results: A total of 63,875 unigenes and 24,154,579 small RNA clean reads were obtained from the roots of P. ginseng. From these small RNAs, 71 miRNA families were identified by bioinformatics target prediction software, including 34 conserved miRNAs, 37 non-conserved miRNA families, as well as 179 target genes of 17 known miRNAs. Through degradome sequencing and computation, we finally verified 13 targets of eight miRNAs involved in transcription, energy metabolism, biological stress and disease resistance, suggesting the significance of miRNAs in the development of P. ginseng. Consistently, major miRNA targets exhibited tissue specificity and complexity in expression patterns.Conclusion: Differential expression microRNAs were found in different growth years of ginsengs(Shizhu ginseng and Yuan ginseng), and the regulatory roles and functional annotations of miRNA targets in P. ginseng need further investigation.

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.

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。

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.

Effects of Raphani Semen on anti-fatigue and pharmacokinetics of Panax ginseng

Objective: To explain the phenomenon that Panax ginseng is not compatible with Raphani Semen based on pharmacodynamics and pharmacokinetics.Methods: The forced swimming time and biochemical parameters such as blood lactate(BLA), serum urea nitrogen(SUN), and hepatic glycogen(GLU) were determined for anti-fatigue experiment. The UPLCMS/MS was used to analyze the pharmacokinetics of Rg1, Re, Rb1, and Rd after orally administration of P. ginseng and P. ginseng combined with Raphani Semen to rats. Pharmacokinetic differences of four ginsenosides between single uses of P. ginseng and combined with Raphani Semen were investigated.Results: The results showed that Raphani Semen tended to significantly reduce the anti-fatigue activity of P. ginseng. Co-administration of P. ginseng and Raphani Semen had significant effects on the pharmacokinetics of the four ginsenosides in rats compared to that observed with P. ginseng extract alone. The AUC0–12 hvalues of the four ginsenosides in PG group were higher than the corresponding values in the PR group. It can be inferred that Raphani Semen decreased the blood exposure of the four ginsenosides in rats when it combined with P. ginseng.Conclusion: The anti-fatigue activity and pharmacokinetic results showed that Raphani Semen may reduce the pharmacological actions of P. ginseng.

人参茎叶挥发油中倍半萜烯化合物的分离与鉴定

对人参茎叶挥发油的有效成分进行分离纯化 ,利用薄层层析 (TL C)、红外光谱 (IR)、气相色谱 (GC)、气相色谱 -质谱 (GC/MS)联机技术从该挥发油中分离鉴定 6个倍半萜烯类化合物 .

高架十字迷路课题中人参根和茎叶皂苷抗焦虑效果的研究

目的 研究人参根和茎叶皂苷抗焦虑效果。方法 高架十字迷路法。结果 中枢兴奋药咖啡因 40～ 80 m g/kg ig给药能使小鼠在开放通路中连续停留时间缩短 ,在封闭通路中停留时间延长。而中枢抑制药安定 1.5～ 3.0mg/ kg却能明显地延长小鼠在开放通路中的停留时间。人参根和茎叶皂苷 5 0～ 2 0 0 m g/ kg ig给药后同样地使小鼠在开放通路中连续停留时间延长 ,并且能使小鼠在开放路中往返次数增加。

复方人参注射液粉末X-射线衍射指纹图谱研究

目的 :建立复方人参注射液的粉末X 射线衍射指纹图谱。方法 :采用粉末X 射线衍射衍射法测定药材、中间体和注射液的衍射图谱 ,对所得到的图谱进行特征峰指认和对比分析 ,获得各自的标准指纹图谱。结果 :10个批次的药材、中间体和注射液均有非常好的重现性 ,相似度达 85 %以上。结论 :粉末X 射线衍射方法可用于复方人参注射液的质量控制。

人参总皂苷诱导K562细胞凋亡的实验研究

目的 研究人参总皂苷诱导白血病细胞凋亡及其机制 ,为进一步开发人参总皂苷提供实验依据。方法 采用细胞体外培养、图象分析技术、流式细胞术、形态学观察与免疫细胞化学等方法 ,研究人参总皂苷诱导 K5 6 2细胞凋亡。结果 人参总皂苷对 K5 6 2细胞有增殖抑制作用 ,并可诱导 K5 6 2细胞凋亡明显增加 ,同时 K5 6 2细胞癌基因产物 C- MYC,BCL- 2表达明显降低。结论 人参总皂苷能诱导 K5 6 2细胞凋亡 ,其机制可能与调控癌基因产物的表达有关。

响应曲面优化超微粉碎参须中人参总皂苷的溶出工艺

目的:研究超微粉碎的粉碎度、提取时间、乙醇体积分数、提取温度等因素对参须中人参总皂苷溶出的影响,并探讨不同因素之间的交互作用。方法:通过单因素试验探讨粉碎度、提取时间、乙醇体积分数、提取温度等因素对参须中人参总皂苷溶出率的影响,在单因素试验基础上,利用响应曲面法对影响人参总皂苷溶出率的3个主要因素即超微粉碎时间、提取时间和乙醇体积分数进行优化。结果:人参总皂苷的含量与所考察的3个因素之间关系符合二次模型。确定最佳溶出工艺参数为:参须超微粉碎时间为9 min,70%乙醇为溶剂,提取温度50℃,提取时间70 min。实验结果显示,此条件下人参总皂苷平均溶出率为94.81%,与模型预测值非常接近,说明该模型比较可靠。结论:该优化工艺溶出快速、高效、简单、稳定。