ent-Labdane diterpenoids from Andrographis paniculata

Six new ent-labdane diterpenoids, 3-O-beta-D-glucopyranosyl-14,19-dideoxyandrographolide (1), 14-deoxy-17-hydroxyandrographolide (2), 19-O-[beta-D-apiofuranosyl(1-->2)-beta-D-glucopyranoyl]-3,14-dideoxyandrographolide (3), 3-O-beta-d-glucopyranosylandrographolide (4), 12S-hydroxyandrographolide (5), and andrographatoside (6), together with 17 known analogues, were isolated from the aerial parts of Andrographis paniculata. The structures of 1-6 were determined by spectroscopic data analysis. All compounds isolated were evaluated for their inhibitory activity against several bacterial and fungal strains.     

Anticancer and immunostimulatory compounds from Andrographis paniculata

Andrographis paniculata extract is traditionally used as a medicine to treat different diseases in India, China and Southeast Asia. In the present study, we evaluated the anticancer and immunomodulatory activity of the methanolic extract of Andrographis paniculata in human cancer and immune cells. The methanolic extract of Andrographis paniculata was fractionated into dichloromethane, petroleum ether and aqueous extracts and screened for bioactivity. Our results indicate that the dichloromethane fraction of the methanolic extract retains the active compounds contributing for both the anticancer and immunostimulatory activity. Dichloromethane fraction significantly inhibits the proliferation of HT-29 (colon cancer) cells and augments the proliferation human peripheral blood lymphocytes (HPBLs) at low concentrations. On further fractionation of the dichloromethane extract we could isolate three diterpene compounds, i.e. [1] andrographolide, [2] 14-deoxyandrographolide and [3] 14-deoxy-11,12-didehydroandrographolide. Andrographolide showed anticancer activity on diverse cancer cells representing different types of human cancers. Whereas all the three molecules showed enhanced proliferation and interleukin-2 (IL-2) induction in HPBLs.     

ent-Labdane diterpenoid lactone stereoisomers from Andrographis paniculata

Two pairs of ent-labdane diterpenoid lactone stereoisomers (1- 4) including three new compounds (1- 3) were isolated from the 85% EtOH extract of the aerial parts of Andrographis paniculata. The structures of these compounds were identified as 7 R-hydroxy-14-deoxyandrographolide (1), 7 S-hydroxy-14-deoxyandrographolide (2), 12 S,13 S-hydroxyandrographolide (3), and 12 R,13 R-hydroxyandrographolide (4) by spectroscopic data analyses and calculated (13)C NMR data at the B3LYP/6-311++G(2d,p)//B3LYP/6-31G* level using the GIAO method. The 12 S-configuration of 4 was revised to 12 R based on the spectroscopic data. The antiproliferative activities of the two pairs of stereoisomers and 14 other ent-labdane diterpenoid derivatives were determined in human leukemia HL-60 cells. Andrographolide (7) and isoandrographolide (12) exhibited higher antiproliferative activities than other ent-labdane diterpenoids with GI 50's of 9.33 and 6.30 microM, respectively.     

Choleretic action of andrographolide obtained from Andrographis paniculata in rats

Andrographolide, a diterpene, isolated from Andrographis paniculata exhibited a strong choleratic action when administered intraperitoneally (i.p.) to albino rats. This substance induced an increase in bile flow together with a change in the physical properties of the bile secretion.     

穿心莲总内酯的NO抑制活性研究

目的：考察穿心莲总内酯对巨噬细胞释放一氧化氮的抑制作用。方法：以脂多糖（LPS）刺激小鼠巨噬细胞RAW264.7 建立体外炎症模型,采用Griess Reagent法检测细胞培养上清一氧化氮（NO）含量并计算抑制率,通过MTT法评价细胞毒性。结果：穿心莲总内酯在 5-50 μmol·L^-1浓度范围内对LPS诱导的小鼠巨噬细胞RAW264.7释放的NO具有明显的抑制作用,并呈现良好的剂量依赖关系,IC50分别为8.58 μmol·L^-1,11.52 μmol·L^-1,8.94 μmol·L^-1;其主要活性成分穿心莲内酯（5-60 μmol·L^-1）、脱水穿心莲内酯（5-100 μmol·L^-1）、新穿心莲内酯（5-100 μmol·L^-1）对LPS诱导的小鼠巨噬细胞RAW 264.7释放的NO亦表现出抑制作用,并呈现良好的剂量依赖关系,其IC50分别为17.54 μmol·L^-1,49.54 μmol·L^-1,41.80 μmol·L^-1。结论：穿心莲总内酯的NO抑制作用优于三个单体,以上结果可为穿心莲总内酯的制剂开发及临床应用提供一定的理论基础及科学依据。     

广东等五省穿心莲药材中穿心莲内酯和脱水穿心莲内酯含量比较

目的通过比较不同产地穿心莲药材中穿心莲内酯和脱水穿心莲内酯含量,寻找穿心莲药材最适宜的栽培地区。方法收集全国5个省9个不同产地25个穿心莲样品,按2010版《中国药典》一部穿心莲项下方法进行检测与分析。结果不同产地及批次穿心莲样品中总内酯的含量在1.03%~6.98%之间;穿心莲不同部位总内酯的含量:叶在3.43%~6.98%之间,茎在0.93%~1.33%之间,全草在1.99%~4.41%之间;不同采集方式穿心莲样品总内酯含量:自采样品含量在3.20%~4.41%之间,产地供应商提供的样品含量在1.99%~3.36%之间。结论穿心莲不同产地、不同采集方式及不同部位的含量均存在着显著差异,选择适宜的产地建立基地、适时采收及加工对穿心莲药材的质量控制意义重大。     

Cytotoxic constituents from Andrographis paniculata induce cell cycle arrest in jurkat cells

Herbal medicines are now attracting attention as potential sources of anticancer agents. Andrographis paniculata is a traditionally used anticancer herb in Indian and Chinese herbal medicine. Phytochemical investigation of the ethanol extract of the aerial parts of this herb resulted in the isolation of 14 compounds including flavonoids and labdane diterpenoids. This is the first isolation of compound 6 from a natural source, and the aerial parts of A. paniculata are a rich source for the molecule andrographolide (9, 1.375%, w/w). The structures of the isolated compounds were established by means of spectral data. The cytotoxic activities of these isolates were evaluated against Jurkat, PC-3, HepG2 and Colon 205 tumor cells, and normal cells PBMCs. The bioactivity assays showed that metabolites 1-4 and 6-8 exhibited moderate cytotoxic activity against Jurkat, PC-3 and Colon 205 cell lines, where compound 6 had IC(50) values of 0.05, 0.07 and 0.05 mm, respectively. Further, among these effective compounds, 3 and 6 selectively blocked the cell cycle progression at G0/G1, while 1, 2, 4, 7 and 8 blocked the same at G2/M phase of the Jurkat cell line. This is the first cell cycle analysis for the above mentioned isolates on the Jurkat cells. Therefore, these plant-derived compounds may play a role in the prevention and/or management of cancer.     

Anti-malarial activity of some xanthones isolated from the roots of Andrographis paniculata

Four xanthones were isolated from the roots of Andrographis paniculata using a combination of column and thin-layer chromatographic methods. They were characterized as (i) 1,8-di-hydroxy-3,7-dimethoxy-xanthone, (ii) 4,8-dihydroxy-2,7-dimethoxy-xanthone, (iii) 1,2-dihydroxy-6,8-dimethoxy-xanthone and (iv) 3,7,8-trimethoxy-1-hydroxy xanthone by IR, MS and NMR spectroscopic methods. In vitro study revealed that compound 1,2-dihydroxy-6,8-dimethoxy-xanthone possessed substantial anti-plasmodial activity against Plasmodium falciparum with its IC(50) value of 4 microg ml(-1). Xanthones bearing hydroxyl group at 2 position demonstrated most potent activity while xanthones with hydroxyl group at 1,4 or 8 position possessed very low activity. In vivo anti-malarial sensitivity test of this compound on Swiss Albino mice with Plasmodium berghei infection using Peters' 4-day test gave substantial reduction (62%) in parasitaemia after treating the mice with 30 mg kg(-1) dose. In vitro cytotoxicity against mammalian cells revealed that 1,2-dihydroxy-6,8-dimethoxy-xanthone is non-cytotoxic with its IC(50) > 32 microg ml(-1).     

穿心莲联用TMP抗菌作用的实验研究

目的：探讨增强中药穿心莲抗菌作用强度的方法。方法：采用琼脂稀释法进行抗菌作用研究。结果：TMP与穿心莲联用对实验6种细菌均有明显的抗菌增效作用。结论：TMP可作为中药穿心莲的抗菌增效剂。     

穿心莲R2R3-MYB基因家族的鉴定与表达分析

R2R3-MYB是一类调控植物生长发育及次生代谢的重要转录因子.利用生物信息学手段从穿心莲全基因组中鉴定R2R3-MYB基因,分析其理化性质、染色体定位信息、基因结构和保守序列信息、系统进化信息及启动子区域的顺式作用元件.结合转录组测序技术和qRT-PCR分析其在非生物胁迫和激素诱导下的表达谱.结果 表明,穿心莲R2R...     

Inhibition of Helicobacter pylori and Its Associate Urease by Labdane Diterpenoids Isolated from Andrographis paniculata

The present study was carried out to evaluate anti‐Helicobacter pylori and its associated urease activity of labdane diterpenoids isolated from Andrographis paniculata. A molecular docking analysis was performed by using ArgusLab 4.0.1 software. The results obtained indicate that compound A possesses strong inhibition to H. pylori, 28 ± 2.98 (minimum inhibitory concentration, 9 µg/mL), and its urease, 85.54 ± 2.62% (IC₅₀, 20.2 µg/mL). Compounds B, C, and D also showed moderate inhibition to H. pylori and its urease. The obtained results were in agreement with the molecular docking analysis of compounds. The phytochemicals under investigation were found to be promising antibacterial agents. Moreover, the isolated compounds can be considered as a resource for searching novel anti‐H. pylori agents possessing urease inhibition. Copyright © 2015 John Wiley & Sons, Ltd.     

In vitro antiprotozoal activity of some xanthones isolated from the roots of Andrographis paniculata

Four xanthones isolated from the roots of Andrographis paniculata were tested in vitro for antiprotozoal activity against Trypanosoma brucei brucei, Trypanosoma cruzi and Leishmania infantum. Compound TDR13011 (1,2-dihydroxy-6,8-dimethoxy-xanthone) showed good activity against T. b. brucei and L. infantum with a 50% inhibitory concentration (IC50) of 4.6 microg/ml and 8 microg/ml respectively. Xanthones from the roots of Andrographis paniculata exhibited promising anti-protozoal activity and these compounds could be chemically modified to obtain a more potent product.     

Analysis on Isozyme and Diterpene Lactones Variability during Ontogenesis of Andrographis paniculata

Objective To understand the relationship between isozyme activities and diterpene lactone biosynthesis of Andrographis paniculata. Methods Plants were collected during ontogeny from seeding to seed maturity, and separated into leaves and stems for determination. Morphological and yield parameters were used to describe plant growing states. Isozyme changes were tested by polyacrylamide gel electrophoresis. HPLC was used to develop the fingerprints as well as to determine the diterpene lactone content. Results Significant increases were observed in the activities of isozymes, such as aspartate aminotransferase(AST), malate dehydrogenase(MDH), peroxidase(POD), and catalase(CAT), around the early stage of bud in leaves, and the activities of these four kinds of isozymes increased gradually as time progressed in stems. The content changes of diterpene lactones in leaves and stem were various. In the leaves, andrographolide(1) was recorded the highest [(23.63 ± 1.06) mg/g] at the early stage of bud, whereas deoxyandrographolide(2) was the lowest [(6.78 ± 0.27) mg/g] at this period and it reached the highest level at the seeding stage [(26.05 ± 1.04) mg/g]. Dehydroandrographolide(3) and neoandrographolide(4) fluctuated during growing stages. Meanwhile, the HPLC fingerprint showed that the content changes of two unknown compounds were related to that of dehydroandrographolide in leaves. In stems, andrographolide had increased gradually until the bud stage [(8.26 ± 0.33)mg/g], and other three diterpene lactones showed a trend of fluctuation. The yield of total diterpene lactones in aerial part reached the highest at the first flowering stage(806.71 mg/plant). Conclusion These results lay the foundation for the future research on the relationship of isozymes and diterpene lactones, and for determining the most favorable time for harvesting A. paniculata.     

密度、磷、氮对穿心莲产量和质量的影响

目的研究密度、磷、氮对穿心莲产量和质量的影响。方法田间随机区组设计,高效液相色谱法测定穿心莲内酯和脱水穿心莲内酯量。结果密度对穿心莲的株高、鲜质量、干质量、脱水穿心莲内酯的量有极显著影响;磷肥对穿心莲内酯量、脱水穿心莲内酯的量、内酯总量等质量性状及经济产量有极显著影响;氮肥对穿心莲的干质量、穿心莲内酯的量、脱水穿心莲内酯的量、内酯总量的经济产量有显著或极显著影响;磷肥,磷、氮互作对经济产量影响较大;鲜质量与干质量间相关极显著;质量性状间、质量性状与经济产量间相关显著或极显著。结论密度1.8×105株/hm2、PO540kg/hm2、纯氮225kg/hm2经济产量最高。     

Antioxidant,antioedema and analgesic activities of Andrographis paniculata extracts and their active constituent andrographolide

Andrographis paniculata (AP), a popular ingredient of Oriental folk medicine, is commonly used for treating infection, inflammation, fever and diarrhoea. In this study, extracts prepared from cultivated AP and their active constituent andrographolide were evaluated for antioxidant, antioedema and analgesic activities. The results showed that the aqueous AP extract (AP‐H₂O) exhibited a greater antioxidant activity than the ethanol AP extract (AP‐EtOH) in all model systems tested. At a concentration of 50 µg/mL, the free radical scavenging, xanthine oxidase inhibition and antilipid peroxidation activities for AP‐H₂O were 66.8%, 57.3% and 65.3%, respectively, and for AP‐EtOH were 57.8%, 52.6% and 34.2%, respectively. At a dosage of 100 mg/kg, AP‐H₂O and andrographolide, but not AP‐EtOH, showed antioedema and analgesic activities. In phytochemical analysis, AP‐H₂O showed a higher concentration of total flavanoid but a lower phenol content than AP‐EtOH. In conclusion, AP‐H₂O was more potent than AP‐EtOH in antioxidant activities. Furthermore, compared with andrographolide, AP‐H₂O as an extract also appears to possess potent antioedema and analgesic activities. Copyright © 2009 John Wiley & Sons, Ltd.     

Protective activity of andrographolide and arabinogalactan proteins from Andrographis paniculata Nees. against ethanol-induced toxicity in mice

To find out the active principles against ethanol-induced toxicity in mice, Andrographis paniculata Nees. (Ap) was chosen and isolated andrographolide (ANDRO) and arabinogalactan proteins (AGPs). ANDRO was detected by HPTLC, FTIR and quantified by HPLC (10mg/g of Ap powder). AGPs was detected by beta-glucosyl Yariv staining of SDS-PAGE gel, FTIR and quantified by single radial gel diffusion assay with beta-glucosyl Yariv reagent (0.5mg/g Ap powder). The mice are pretreated intra-peritoneally (i.p.) with different doses (62.5, 125, 250, and 500mg/kg) of body weight of mice] of ANDRO and AGPs for 7 days and then ethanol (7.5g/kg of body weight) was injected, i.p. Besides, silymarin was used as standard hepatoprotective agent for comparative study with ANDRO and AGPs. The ameliorative activity of ANDRO and AGP against hepatic renal alcohol toxicity was measured by assessing GOT, GPT, ACP, ALP and LP levels in liver and kidney. It has been observed that pretreatment of mice with ANDRO and AGPs at 500mg/kg of body weight and 125mg/kg of body weight respectively could able to minimize the toxicity in compare to ethanol treated group as revealed by the different enzymatic assay in liver and kidney tissues and the results were comparable with silymarin. Hence, out of several ill-defined compounds present in Ap, ANDRO and AGPs are the potential bioactive compounds responsible for protection against ethanol-induced toxicity.     

The screening of extracts from Goniothalamus scortechinii,Aralidium pinnatifidum and Andrographis paniculata for anti-malarial activity using the lactate dehydrogenase assay

Goniothalamus scortechinii, Andrographis paniculata and Aralidium pinnatifidum were selected for the study based on their ethnomedicinal values. They were screened for anti-malarial activity towards Plasmodium falciparum in vitro using the lactate dehydrogenase (LDH) assay. The crude extract of G. scortechinii exhibited the most potent schizonticidal activity compared to the other extracts. It is effective against both the chloroquine resistant isolate, Gombak A and the sensitive strain, D10 of Plasmodium falciparum. Furthermore a better IC(50) value was obtained against the resistant strain, (9 microg/ml) compared to the sensitive strain, 40 microg/ml. When the crude extract was fractionated into 3 fractions, the chloroform fraction yielded the best activity, exhibiting equipotency against both strains of parasite used; IC(50) of 23.53 microg/ml against Gombak A and 21.06 microg/ml against D10.     

不同产地穿心莲药材色谱指纹图谱的比较研究

目的:建立穿心莲药材的HPLC指纹图谱,为其全面质量评价及控制提供参考。方法:采用HPLC/UV法,测定了10个不同产地的23批穿心莲药材样品。色谱条件:Shimadzu ODS柱,流动相为0.1%甲酸乙腈与0.2%甲酸,梯度洗脱,流速1.0 ml/min,检测波长254 nm,柱温为室温。应用聚类分析、非线性映射和相似度计算等方法对所得指纹图谱进行了定性、定量评价。结果:建立了操作简单、分离度佳、重现性好的穿心莲药材指纹图谱检测方法,不同产地药材指纹图谱具有一定差异。结论:聚类分析、非线性映射和相似度计算等方法可实现对指纹图谱的定性、定量评价,所建立的色谱指纹图谱可用于穿心莲药材的鉴别及质量评价。     

Role of semi-purified andrographolide from Andrographis paniculata extract as nano-phytovesicular carrier for enhancing oral absorption and hypoglycemic activity

Objective: Andrographis paniculata is a well-known medicinal plant in Southeast Asia, India and China.The plant contains andrographolide(AN), a very important phytochemical used in various health problems. However, AN is low in oral absorption bioavailability of AN due to the rapid clearance and high protein binding capacity.Methods: The present study was aimed to develop a nano-phytovesicular formulation of semi-purified AN extracts from a naturally occurring phospholipid(soya phosphatidylcholine) in order to increase the oral absorption and antihyperglycemic activity in rats.Results: The nano-phyto vesicle of semi-purified AN extracts equivalent to 25 mg/kg AN significantly protected the hyperglycemic condition of rats. The in vitro and in vivo experiments results proved that the nano-phytovesicular system of plant extracts containing AN produced better oral absorption, bioavailability and improved antihyperglycemic activity compared with that of free AN at dose of 50 mg/kg.Conclusion: Hence, the prepared semi-purified extract nano-phytovesicular system is helpful in solving the problem of rapid clearance of AN.     

Antiviral properties of ent-labdene diterpenes of Andrographis paniculata nees, inhibitors of herpes simplex virus type 1

Andrographolide, neoandrographolide and 14-deoxy-11,12-didehydroandrographolide, ent-labdene diterpenes isolated from Andrographis paniculata showed viricidal activity against herpes simplex virus 1 (HSV-1). None of these compounds exhibited significant cytotoxicity at viricidal concentrations.