Inhibition of prostaglandin E2 production by platycodin D isolated from the root of Platycodon grandiflorum.

Platycodin D, isolated from the root of Platycodon grandiflorum A. DC. (Campanulaceae) suppressed prostaglandin E2 production at 10 and 30 microM in rat peritoneal macrophages stimulated by the protein kinase C activator 12-O-tetradecanoylphorbol 13-acetate (TPA). Platycodin D3 and oleanolic acid showed no effect at these concentrations. Western blot analysis revealed that the induction of COX-2 protein by TPA was inhibited by platycodin D in parallel with the inhibition of prostaglandin E2 production. Platycodin D showed no direct effect on COX-1 and COX-2 activities. TPA-induced release of [3H]arachidonic acid from pre-labeled macrophages was also not inhibited by platycodin D.     

Isolation of a new saponin and cytotoxic effect of saponins from the root of Platycodon grandiflorum on human tumor cell lines

A novel triterpenoid saponin, deapioplatycoside E (1) was isolated from the root extract of Platycodon grandiflorum, together with the seven known saponins 2 - 8, i. e., platycoside E (2), deapioplatycodin D3 (3), platycodin D3 (4), polygalacin D2 (5), platycodin D2 (6), deapioplatycodin D (7) and platycodin D (8). The structure of the new saponin 1 was determined on the basis of spectral analysis and chemical evidence. The crude saponin fraction (ED50: ca. 10 - 15 microg/mL) and compounds 6 - 8 (ED50: ca. 4 - 18 microg/mL) exhibited significant inhibition on the proliferation of five kinds of cultured human tumor cell lines, i. e., A549 (non-small cell lung), SK-OV-3 (ovary), SK-MEL-2 (melanoma), XF498 (central nerve system) and HCT-15 (colon), in vitro.     

反相高效液相色谱法测定桔梗中3种三萜皂苷类化合物的含量

目的建立RP-HPLC法同时测定桔梗中去芹糖桔梗皂苷E、桔梗皂苷E和桔梗皂苷D3的含量。方法以Hypersil-ODS(4.6 mm×250 mm,5μm)为色谱柱,流动相为乙腈(A)-水(B),梯度洗脱,流速为0.8 mL.min-1,检测波长210 nm,温度为室温。结果去芹糖桔梗皂苷E、桔梗皂苷E和桔梗皂苷D3分别在4.160~83.2、10.75~215.0、4.080~81.6μg.mL-1与峰面积线性关系良好,平均回收率分别为96.5%、97.0%和97.3%,RSD分别为1.2%、1.0%和1.3%。结论该方法快速稳定且分离度好,为桔梗的质量控制提供可借鉴的分析方法。     

Platycodin D and D3 isolated from the root of Platycodon grandiflorum modulate the production of nitric oxide and secretion of TNF-alpha in activated RAW 264.7 cells

Platycodon D (PD) and D3 (PD3) isolated from Platycodon grandiflorum has been previously reported to show anti-inflammatory activities in rats. In this study, the production of proinflammatory cytokines, nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) was examined in a macrophage like cell line, RAW 264.7 cells, in the presence of PD and PD3, oligosaccharide derivatives of oleanolic acid. RAW 264.7 cells activated with lipopolysaccharide (LPS; 1 microg/ml) and recombinant interferon-gamma (rIFN-gamma; 50 U/ml) were treated with various doses of PD and PD3 for 24 h. Supernatants were analyzed for the production of NO and TNF-alpha using Griess reagent and enzyme-linked immunosorbent assay (ELISA), respectively. NO was inhibited in a dose-dependent manner by PD and PD3 (IC50 of platycodin D approximately 15 uM, IC50 PD3 approximately 55 uM). The expression of inducible NOS (iNOS) was inhibited by these compounds, as measured by Western blot analysis, as well as the expression of iNOS mRNA, as measured by Northern blot analysis. RAW 264.7 cells were treated at various times after LPS and activation with PD. Treatment with PD up to 8 h after activation showed significant inhibition of NO, indicating that early signal transduction of NOS synthesis may be inhibited by PD. In contrast to NO, secretion of TNF-alpha as well as expression of TNF-alpha mRNA was increased by PD and PD3. TNF-alpha secretion from RAW 264.7 cells was measured at various times after LPS and rIFN-gamma activation. Secretion of TNF-alpha was also increased up to 8 h postactivation, suggesting that PD may stimulate TNF-alpha synthesis or inhibit degradation of TNF-alpha mRNA. Oleanolic acid was without effect on both the production of NO and secretion of TNF-alpha. These data suggest a dichotomous regulation of these important proinflammatory mediators by PD and PD3.     

桔梗同源四倍体株系的鉴定和桔梗皂甙D含量的比较

对组织培养条件下诱导获得的桔梗(Platycodon grandiflorum A．DC)四倍体株系，进行了农艺性状的鉴定，同时采用HPLCELSD测定了其桔梗皂甙D的含量。结果表明：在供试的21个桔梗四倍体株系中，18个株系的桔梗皂甙D含量高于二倍体株系；9个株系的单株产量高于二倍体株系；15个株系的桔梗皂甙D的生产量高于二倍体株系。这一结果为今后进一步选育出产量高、皂甙含量高的优良桔梗品种展示了美好前景。     

Lead compounds for anti-inflammatory drugs isolated from the plants of the traditional oriental medicine in Korea

Effects of compounds isolated from medicinal plants in Korea on prostaglandin E(2) (PGE(2)) production in rat peritoneal macrophages were examined, and mechanism of action of the active constituents was analyzed. The active constituents were as follows; tectorigenin and tectoridin isolated from the rhizomes of Belamcanda chinensis, platycodin D isolated from the roots of Platycodon grandiflorum, imperatorin isolated from the roots of Angelica dahurica, and hyperin isolated from the roots of Acanthopanax chiisanensis. These compounds inhibit the induction of cyclooxygenase-2 (COX-2), thus inhibiting PGE(2) production. The chemically synthesized chalcone derivative, 2'-hydroxy-4'-methoxychalcone, also inhibits PGE(2) production by suppressing COX-2 induction. In contrast, taiwanin C isolated from the roots of Acanthopanax chiisanensis inhibited PGE(2) production by direct inhibition of COX-1 and COX-2.     

Evaluations of saponin properties of HPLC analysis of Platycodon grandiflorum A.DC

Platycodon root, one of the most important Chinese herbal medicines, has been used as an antiphlogistic, antitusivie, and expectorant agent since ancient times. In the Japanese Pharmacopoeia XIV this is listed as the root of Platycodon grandiflorum A. De Candolle (Campanulaceae) and called KIKYOU (Platycodi Radix) in Japanese. HPLC analysis showed that commercial samples of P. Radix all contained platycodins, and a total of 12 peaks were identified by co-HPLC analysis with authentic samples isolated earlier from this laboratory. The peak purity and identity were checked with a photodiode array detector. The contents of the major saponins, platycodins A, C, and D, were determined and the peak-area ratios of platycodins A, C, and D, were shown to be correlated with their sources of origin. Fourteen commercial samples of Platycodon root, the origin of which was Platycodon grandiflorum, were collected from China (5 samples), Korea (5 samples), and Japan (4 samples). The commercial samples from China, Korea, and Japan each gave a distinct HPLC pattern with peak-area ratio of platycodins A, C, and D, of 1:2:3 and 2:8:1, respectively. HPLC analysis showed that those on the Japanese market were either imported from China or Korea based upon their HPLC patterns.     

Activation of mitogen-activated protein kinases and AP-1 by polysaccharide isolated from the radix of Platycodon grandiflorum in RAW 264.7 cells

The root of Platycodon grandiflorum has been widely used for the treatment of various diseases in oriental medicine. Our previous study showed that the PG, a polysaccharide isolated from P. grandiflorum, activates macrophages via Toll-like receptor 4 (TLR4). However, the associated biological mechanisms are not fully understood. To elucidate the molecular mechanism responsible for the macrophage activation, we investigated the effect of PG on the activity of mitogen-activated protein kinases (MAPKs) and activator protein-1 (AP-1) in RAW 264.7 cells, a murine macrophage cell line. Treatment of RAW 264.7 cells with PG produced a marked induction of AP-1 DNA binding activity. Moreover, all three MAPKs were activated by PG, and PG-induced activation of MAPKs was abrogated by the treatment of PD98059, curcumin, and SB203580, specific inhibitors of MEK-1/2, stress-activated protein kinases/jun N-terminal kinase (SAPK/JNK), and p38 MAP kianse, respectively. The induction of AP-1 DNA binding activity by PG was also inhibited by these MAPK inhibitors. Moreover, supershift analysis identified that JunB and Fra-1 are major components involved in the PG-mediated induction of AP-1 DNA binding. Additionally, curcumin and SB203580 suppressed PG-induced production of nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha), whereas PD98059 showed an inhibitory effect only on the TNF-alpha production. Taken together, these results suggest that macrophage activation by PG is mediated, at least in part, by MAPKs and AP-1.     

Anti-atherosclerotic Activity of Platycodin D Derived from Roots of <i>Platycodon grandiflorum</i> in Human Endothelial Cells

This study examined the effects of platycodin D (PD), a triterpene saponin from the the root of Platycodon grandiflorum A.DC on human umbilical vein endothelial cells (HUVECs) in vitro, which were pre-treated with PD (0.01, 0.15, 0.25?mg/mL), respectively, and treated with 50?mg/L oxidized low-density lipoprotein (oxLDL). The levels of nitric oxide (NO) and malonaldehyde (MAD) in the culture medium, vascular cell adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) mRNA expression in endothelium cells and the adhesion of monocytes to endothelial cells were measured. The results showed that PD increased NO concentration and decreased MDA level induced by oxLDL in the medium of endothelial cells. Moreover, PD significantly inhibited the oxLDL-induced increase in monocyte adhesion to endothelial cells as well as decreasing mRNA expression levels of VCAM-1 and ICAM-1 on these cells. Based on these results, it is suggested that PD is a promising anti-atherosclerotic activity, which is at least in part the result of its increasing NO concentration, reducing the oxLDL-induced cell adhesion molecule expression in endothelial cells and the endothelial adhesion to monocytes.     

天王补心丸全方及方中缺桔梗对小鼠镇静催眠作用的影响

目的:探讨中药古方天王补心丸对小鼠中枢神经系统的药理影响及舟楫之剂桔梗在该方剂中的作用.方法:采用小鼠自发活动计数、延长戊巴比妥钠睡眠时间及抗惊厥试验等方法,观察天王补心丸方、缺桔梗的天王补心丸方、天王补心丸中臣药酸枣仁、酸枣仁与桔梗配伍及其桔梗单独使用时的镇静、催眠与抗惊厥作用.结果:天王补心丸有良好的镇静、催眠与抗惊厥作用,但该方去掉桔梗后,镇静、催眠与抗惊厥效果明显下降;天王补心丸中重要成分酸枣仁本身有较好的镇静催眠作用加上桔梗后效果得到加强,但不如天王补心丸全方,桔梗本身没有镇静、催眠与抗惊厥作用.结论:在天王补心丸方剂的配伍中,桔梗作为舟楫之剂确实能载药上浮,增强该方对中枢神经系统的作用.     

Effects of baicalein,berberine, curcumin and hesperidin on mucin release from airway goblet cells

Baicalein, berberine, curcumin and hesperidin are the major components derived from Scutellaria baicalensis, Coptis japonica, Curcuma longa and Poncirus trifoliata, respectively. These plants have been used for the treatment of diverse chronic inflammatory diseases including respiratory disease in oriental medicine and their respective major components were reported to have various biological effects including anti-inflammatory activity. In the present study, we investigated whether these four natural products affect mucin release from airway goblet cells and compared the possible activities of these agents with the inhibitory action on mucin release by PLL and the stimulatory action by ATP. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled using 3H-glucosamine for 24 h and chased for 30 min in the presence of varying concentrations of each agent to assess the effects on 3H-mucin release. The results were as follows: (i) baicalein did not affect mucin release significantly; (ii) berberine, curcumin and hesperidin increased mucin release at the highest concentration (10 - 4 M); (iii) PLL inhibited and ATP increased mucin release. We conclude that berberine, curcumin and hesperidin can increase mucin release by directly acting on airway mucin-secreting cells and suggest that these agents be further studied for possible use as mild expectorants during the treatment of chronic airway diseases. Abbreviations. PLL:poly- L-lysine ATP:adenosine triphosphate HTSE:hamster tracheal surface epithelial DMSO:dimethylsulfoxide IL-12:interleukin-12 PBS:phosphate-buffered saline     

桔梗的药理学研究进展

桔梗在药理方面的作用有镇咳、祛痰、抗炎、保肝、抗癌等药理活性。桔梗作为一种药食两用的植物,具有广阔的开发应用前景。本研究通过查阅近些年关于桔梗研究的文献,对桔梗药理活性的新进展进行了系统的总结,为桔梗在药理方面的研究和开发利用提供科学依据。     

不同加工方法对桔梗根中多糖含量的影响

目的了解不同加工方法对桔梗多糖含量的影响。方法采用苯酚-硫酸法测定多糖的含量。结果不同干燥方法间桔梗多糖含量差异显著,依次为：切片干燥〉去皮干燥〉带皮干燥;桔梗鲜根收获后多糖含量随着放置时间的延长显著降低;鲜根放置去皮与否对多糖含量无显著影响。结论为保证桔梗根中多糖含量,桔梗鲜根收获后应及时切片烘干。     

天王补心丸中丹参与桔梗配伍后的化学成分变化研究

目的研究天王补心丸中丹参与桔梗两味药共同煎煮时所产生的化学成分变化情况。方法分别制备桔梗单煎液、丹参单煎液、丹参和桔梗混煎液、丹参和桔梗单煎后混合液4个样品,采用高效液相色谱（HPLC）法分别建立色谱图并比较其差异,测定样品中丹参素钠的含量。结果丹参和桔梗混煎液的色谱峰数目明显多于丹参和桔梗单煎后混合液,丹参素钠的含量前者为0．20％,后者为0．11％。结论丹参与桔梗配伍后,共同煎煮过程中化学成分发生了一些变化,不等同于单味药材煎煮后化学成分的简单相加。     

Effect of processing method on platycodin D content in <Emphasis Type="Italic">Platycodon grandiflorum</Emphasis> roots

Platycodon grandiflorum root is a traditional medicine and food material rich in triterpenoid saponins. Its major constituent, platycodin D (PD), is known to have various pharmacological properties, but processing methods may influence the PD content. In this study, a fully validated HPLC–ELSD method was developed for the quantification of PD in various states of 73 P. grandiflorum root samples from East Asia, and it exhibited a marked variation of the content. Furthermore, the effects of processing procedures such as peeling and drying temperature on the PD content were investigated using UPLC–ELSD analysis, and as a result, a significant influence of processing methods such as peeling and heating of samples on the content was confirmed. Specifically, unpeeled samples that were dried at 40 °C showed the greatest PD content. The obtained results could facilitate the reliable standardization of P. grandiflorum for precise authentication and efficacious applications.     

Cholesterol-lowering effect of platycodin D in hypercholesterolemic ICR mice

This study investigates the in vivo hypocholesterolemic action of platycodin D and its in vitro evidence for the cholesterol-lowering properties. In order to examine the effects of platycodin D on hypercholesterolemia in male ICR mice, platycodin D with doses of 15, 30 or 50 mg/kg was orally administered for 8 weeks. Changes in body weight and daily food intake were measured regularly during the experimental period. Final contents of triglyceride and different types of cholesterol in the serum, livers and feces were determined. The effects of platycodin D on cholesterol metabolism were further investigated with several in vitro assays, including antioxidant effect on low density lipoprotein oxidation, inhibition of human acyl-coenzyme A:cholesterol acyltransferase (hACAT) and serum lipoprotein associated-phospholipase A(2) (Lp-PLA(2)), as well as the regulation of farnesoid X receptor. The formation of insoluble complex between platycodin D and cholesterol was also investigated. Following an eight week experimental period, the body weights of platycodin D-fed mice were less than those of control mice on a high cholesterol diet by 11.2+/-5% (P<0.01) with 15 mg/kg platycodin D, 11.7+/-5% (P<0.01) with 30 mg/kg platycodin D, and 23.4+/-7.9% (P<0.0001) with 50 mg/kg platycodin D, respectively. A decrease in daily food consumption was also noted in most of the treated animals. Triglyceride and cholesterol concentrations were decreased in serums and livers, but increased in feces. Some of the in vitro observations revealed that the hypocholesterolemic effect of platycodin D is partly associated with inhibition to hACAT activity and antagonism to the farnesoid X receptor as well as the formation of insoluble complex with between platycodin D and cholesterol. Both in vivo and in vitro results demonstrate a potential value of platycodin D as a novel cholesterol-lowering and anti-atherogenic candidate.     

桔梗中远志酸型皂苷的化学研究

目的分离、鉴定桔梗［Platycodon grandiflorum (Jacq.)A.DC］根中的远志酸型皂苷类化学成分。方法 采用乙醇提取、乙酸乙酯萃取，大孔树脂柱色谱、硅胶柱色谱及高效液相色谱等方法进行分离，得到3个化学成分，运用IR，MS，¹H NMR和¹³C NMR等光谱法鉴定化合物的结构。结果分离鉴定了3种三萜皂苷：3-O-β-D-laminaribiosyl polygalacic acid (I)，3-O-β-D-glucopyranosyl polygalacic acid (II)及polygalacin D (III)。结论化合物I是新化合物，II，III为已知化合物。化合物II为首次从该植物中分得，化合物I和II也是首次从桔梗中分离得到的单糖链糖苷。     

中药桔梗对丝裂霉素诱发小鼠骨髓微核率的影响

[摘 要] 目的：观察中草药桔梗对丝裂霉素（MMC）诱发小鼠微核的抑制作用。方法： 50只小鼠随机分为阴性对照组（N?S）,阳性对照组（MMC,30mg? kg-1）;桔梗水煎液诱变组（1.0、2.0、4.0和8.0 g?kg-1 ,相当于人5×、10×、20×、40×临床常用剂量）; 桔梗水煎液抗诱变组（MMC 0.4mg?kg-1＋桔梗各剂量组）。每组5只动物,雌雄兼用。MMC腹腔注射给药,其他药物均为灌胃给药,抗诱变实验组桔梗和MMC同时给药,小鼠骨髓嗜多染红细胞微核实验按常规方法进行。结果： 桔梗各剂量组所诱导的小鼠微核频率与正常对照组比较,差异无显著性(P>0.05)。桔梗本身不能诱发小鼠微核,并且4.0g.kg-1,8.0g.kg-1组与丝裂霉素一同给药时,可使丝裂霉素所诱发的小鼠微核率明显降低(P＜0.05)。结论：桔梗对遗传物质无诱变作用,对小鼠遗传物质具有一定的保护作用。     

桔梗对大鼠血浆中异烟肼浓度的影响

目的:研究桔梗提取液对大鼠血浆中异烟肼浓度的影响。方法:以异烟肼单独给药作为对照组,以异烟肼与桔梗提取液同时给药作为实验组,采用衍生化法分别测定对照组与实验组不同时间点大鼠的血药浓度。结果:实验组与对照组均在灌胃1.0h达到最大血药浓度,实验组血药浓度比对照组高出(65.8±1.05)%(n=5,P<0.05)。结论:桔梗与异烟肼合用后可显著提高异烟肼在大鼠体内的血药浓度。     

桔梗的化学成分和抗肿瘤活性研究进展

桔梗作为一种药食两用的植物,具有显著的镇咳祛痰、抗肿瘤、抗氧化及降血糖等作用。迄今为止,从桔梗中共分离得到55种皂苷类化合物,三萜皂苷是其主要的活性成分。本文对近年来桔梗的化学成分研究进展进行了总结,并从诱导肿瘤细胞凋亡、增强机体免疫功能、抑制端粒酶活性、抑制肿瘤细胞浸润与转移以及阻滞细胞周期等方面,对国内外桔梗抗肿瘤活性的研究进展进行了阐述。