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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.
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The anti-venom activity of Andrographis paniculata (Burm.f.) Nees roots (APR) dichloromethane crude extractsand a promising APR constituent, skullcapflavone I (SKI) was investigated by monitoring the inhibition of secretoryphospholipase A2 (sPLA2) of Naja philippinensis Taylor venom (NPV) crystallized samples. Gas chromatographymass spectrometry was used for the characterization of extracts, while molecular docking was utilized to understandanti-venom properties. Chromatographic analyses primarily revealed the presence of methoxylated flavones. NPV wasfound to have sPLA2 activity (0.0796 ± 0.0018 μmol/minutes/ml) that has been attributed to the poisonous effects.SKI (IC50: 51.1 ± 3.5 μg/ml), isolated from APR showed strong inhibitory effect on phospholipase activity comparedwith dichloromethane extracts of APR (IC50: 192.7 ± 10.9 μg/ml) indicating that SKI was the cause of the bioactivityin APR. Molecular docking simulations showed corresponding results with highly negative binding energies (−6.59 to−8.72 kcal/mol) predicted for the binding of SKI to PLA2 proteins. An important trend found was the presence of freebound Ca2+ lowered binding energies signifying that Ca2+ a has role in the binding of the SKI to PLA2 proteins. Theanti-venom property of APR and the pure compound SKI, upon further studies, could be the first line of defense in themedical protocol of snake venom neutralization.
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The anti-venom activity of Andrographis paniculata (Burm.f.) Nees roots (APR) dichloromethane crude extractsand a promising APR constituent, skullcapflavone I (SKI) was investigated by monitoring the inhibition of secretoryphospholipase A2 (sPLA2) of Naja philippinensis Taylor venom (NPV) crystallized samples. Gas chromatographymass spectrometry was used for the characterization of extracts, while molecular docking was utilized to understandanti-venom properties. Chromatographic analyses primarily revealed the presence of methoxylated flavones. NPV wasfound to have sPLA2 activity (0.0796 ± 0.0018 μmol/minutes/ml) that has been attributed to the poisonous effects.SKI (IC50: 51.1 ± 3.5 μg/ml), isolated from APR showed strong inhibitory effect on phospholipase activity comparedwith dichloromethane extracts of APR (IC50: 192.7 ± 10.9 μg/ml) indicating that SKI was the cause of the bioactivityin APR. Molecular docking simulations showed corresponding results with highly negative binding energies (−6.59 to−8.72 kcal/mol) predicted for the binding of SKI to PLA2 proteins. An important trend found was the presence of freebound Ca2+ lowered binding energies signifying that Ca2+ a has role in the binding of the SKI to PLA2 proteins. Theanti-venom property of APR and the pure compound SKI, upon further studies, could be the first line of defense in themedical protocol of snake venom neutralization.
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Objective To identify the active components of Andrographis paniculata coupled with Caco-2 cells screened by UPLC-Q-TOF-MS. Methods The cell-combining compounds from A. paniculata were screened based on drug uptake and transport absorption experiment using Caco-2 cells, then the cell dissociation solution was detected before and after treating with A. paniculata by using UPLC-Q-TOF-MS method. The cell-combining compounds from the ethanol extracts of A. paniculata were identified based on the retention time and mass spectrometry information of each chemical composition combined with comparing the extracted ion chromatograms and mass spectrometry datas of reference substances and the related articles. Results A total of 10 compounds combined with Caco-2 cells in ethanol extract of A. paniculata, which were detected and identified as 12,13- dihydroandrographolide (1), skullcapflavone I-2'-O-glucoside (2), andrographolide (3), 14-deoxy-11,12-didehydro andrographiside (4), 7-hydroxy dehydroandrographolide (5), neoandrographolide (6), 3-deoxy dehydroandrographiside (7), deoxyandrographolide (8), dehydroandrographolide (9), and one unidentified component. Conclusion This research establishes a method of screening the active components of A. paniculata coupled with Caco-2 cells using UPLC-Q-TOF-MS analysis, which can be used to screen out the active components of the complex system on traditional Chinese medicines, as well as to lay the foundation for further study of the synergetic compatibility effects among active ingredients.
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Objective To study the bitterness inhibition law of hydroxypropyl-β-cyclodextrin (HP-β-CD) concentration (C) on the bitter compounds and bitter Chinese herbal medicine, and to explore the application of electronic tongue in this study. Methods Berberine, oxymatrine, Sophora flavescens, and Andrographis paniculata decoction were used as bitterness vectors to establish two models of bitterness inhibition law about ΔI-C and ΔIe-C, and to explore the prediction model of bitterness inhibition effect about ΔI-ΔIe, based on the oral taste evaluation results (ΔI) and electronic tongue information (ΔIe). Then, fitting precision and fitting goodness of the prediction model were evaluated with cross-validation and residual analysis. Results In this study, good Weibull model of bitterness inhibition pattern about ΔI-C were established for all the four bitterness vectors, the decision coefficient R2 are as followed: 0.999 6, 0.987 9, 0.996 4, and 0.998 4 (P < 0.01); The decision coefficient R2 of six (two sensors per vector) models of bitterness inhibition law about ΔIe-C of berberine, S. flavescens, and A. paniculata decoctions were as followed: 0.996 5, 0.991 6, 0.997 3, 0.989 3, 0.999 6, and 0.999 1 (P < 0.01); The decision coefficient R2 of six corresponding linear prediction models of bitterness inhibition effect about ΔI-ΔIe were as followed: 0.989 1, 0.968 3, 0.989 0, 0.982 0, 0.977 9, and 0.986 1 (P < 0.01); The correlation coefficient R calculated by correlation coefficient of six prediction models above were as followed: 0.986 0, 0.997 3, 0.988 4, 0.960 8, 0.980 2, and 0.983 9 (P < 0.01); No model was established for oxymatrine within the range of tested concentration in this research, as it didn’t respond to the four sensors with varied concentration. Conclusion Based on this method, the bitterness inhibition law of HP-β-CD with changed concentration was obtained. Prediction models based on HP-β-CD concentration or electronic tongue data were also established, they can be used to predict the relative bitterness inhibition effect. Part of the bitter compounds didn't response to the electronic tongue regularly, remain further research and development of electronic tongue technology.
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Objective: To investigate the correlation of water and different volume fractions of ethanol boiling point and saturated steam pressure when using the vacuum extraction technology, and to compare the impact on saturation steam pressure of the addition of different medicine materials (Andrographis paniculata, Carthamus tinctorius, Angelica dahurica, Uncaria rhynchophylla, Scutellaria baicalensis, Salvia miltiorrhiza, Citrus reticulata) or the addition of materials with different particle sizes. Methods: To obtain the theoretical value and measured value of saturated steam pressure under different boiling points by the combined method of theoretical calculation, experiment measurement, and statistical analysis Results: The regression equation for the saturated steam pressure (P) with the volume fraction of ethanol solvent (V) and the boiling point (T) was P = 76.467 1+0.035 2 T2+1.201 0 TV-30.749 4 V2-3.123 9 T-14.966 7 V. The results showed that measured values of the saturated steam pressure of pure solvent, water and different concentration of ethanol, was less than the theoretical value. After adding herbs, the measured value of saturated steam pressure was higher than the pure solvent. The saturated steam pressure would reduce exactly as the extraction temperature reaches a certain temperature after the herbs were added. Conclusion: The addition and crush of herbs have influence on the saturated vapor pressure of solvent with small range.
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Objective: Andrographolide is the main bioactive substance in Andrographis paniculata, and popularly used as active pharmaceutical ingredient (API). We have cloned the gene encoding geranylgeranyl diphosphate synthase from A. paniculata and characterized its tissue expression pattern. Methods: Total RNA was extracted with CTAB-LiCl extraction method; Conserved fragment was amplified and cloned with degenerated primers, and a full length ORF encoding geranylgeranyl diphosphate synthase was obtained with RACE method and analyzed by bioinformatic softwares, e.g. ProtParam. Tissue expression pattern was predicted with real time PCR. Results: We have cloned a 1 047 bp GGPS gene encoding a sequence with 348 amino acids. This amino acid sequence contained a plastid targeted N-terminal signal peptide and has high similarities with the GGPS protein from Catharanthus roseus. The GGPS gene has expressed in a dynamic state in stems and leaves of A. paniculata. The expression reached a high level at bud stage, then decreased at early flowering stage, increased at flowering and early seed setting stage again, and finally decreased at seed setting stage. Considering the above expression characteristics, biosynthesis of metabolites regulated by GGPS was deduced more active at bud stage and flowering and early seed setting stage. Conclusion: GGPS is a key enzyme in biosynthesis of andrographolide. We have cloned the GGPS gene from A. paniculata, and provide a sharp tool in genetic engineering of andrographolide biosynthsis pathway.
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OBJECTIVE: To establish the relationship between the physical parameters of granules and their dissolution behavior. METHODS: Andrographis paniculata (Burm. f.) Nees extract was utilized as a model drug. The extract was mixed with different excipients (MCC/soluble starch) at the ratio of 1:1 by weight using different methods. The granules were prepared by using high shear granulation and extrusion granulation methods. Then the dissolution behavior of the granules was evaluated by monitoring equilibrium dissolution quantity and dissolution rate constant. The effect of physical properties on the dissolution behavior was analyzed through stepwise regression. RESULTS: Particle size, critical relative humidity (CRH), moisture content, pore volume, and tapped density were the main factors related to the dissolution behavior of granules. CONCLUSION: A higher equilibrium dissolution quantity could be obtained with higher moisture content, smaller particle size, and lower CRH. There is a positive correlation between dissolution rate constant and tapped density of granules, and a negative correlation with particle size (D[4, 3]) and pore volume (PV).
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Objective: To study the dynamic changes of the active components from Andrographis paniculata at various growth periods grown in Suixi in Guangdong province, Zhangpu in Fujian province as well as introduced and cultivated in Nanjing and Huaian in Jiangsu province, and to investigate the expression level of the key enzyme gene at the same time in order to obtain the accumulation trend of the active contents from A. paniculata and explore a reasonable harvest and its molecular mechanism. Methods: The contents of andrographolide and dehydroandrographolide were analyzed by HPLC. The expression of key enzyme gene, ent-copalyl diphosphate synthase (CPS) was measured at mRNA level with QRT-PCR. Results: The growth periods of A. paniculata were similar in different areas. It grew fast at vegetative stage while slowly at reproductive stage. The contents of active ingredients in A. paniculata from four areas showed the same change tendency. The content of andrographolide reached the highest level at flowering period. At the same time, the changes of dehydroandrographolide was flatting out. The expression of CPS reached its maximum at bud to flowering period. There was a significant positive correlation between the content of andrographolide and the expression of CPS. Conclusion: The optimal harvest time of A. paniculata should be judged on phenological period and the best harvest period is flowering period.
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Objective: To investigate the intestinal absorptive characteristics of andrographolide (A) and dehydroandrographolide (DDA) in the extract of Andrographis paniculata in rats. Methods: The intestine of rat was cannulated for in situ perfusion. UV was used to determine the concentration of phenol red, and HPLC was used to determine the concentration of A and DDA. The effects of drug concentration, pH value, and absorption site on the absorption had been studied. Results: In 111.22-335.78 μg/mL dose range of A. paniculata extract, Ka and P of A were reduced with the increase of concentration. With the pH value increasing from 5.34 to 6.38, Ka and P were increased slightly. With the pH value increasing from 6.38 to 7.40, Ka and P had a downward trend. A was absorbed in various sections of intestines, and the P descended in the order of duodenum, jejunum, ileum, and colon. In 111.22-222.78 μg/mL dose range of A. paniculata extract, Ka and P were reduced with the increase of concentration. In 222.78-335.78 μg/mL dose range, Ka and P had no change with the increase of concentration. Under the condition of pH 5.34, the absorption was better. With the pH value increasing from 5.34 to 7.40, Ka and P showed a slight downward trend, and P descended in the order of ileum, jejunum, colon, and duodenum. Conclusion: The intestinal absorption mechanism of A and DDA in A. paniculata extract is not just passive transport, but also including the carrier medium transport which are both affected by pH value. The best absorption site of A is duodenal, and the best absorption site of DDA is ileumother. Other components may have no significant influence to the intestinal absorption of A, but they may promote the intestinal absorption of DDA.
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[ Objective ] To supply a technological evidence for the rapid reproduction of Andrographis paniculata (Burm. f.) Nees (APN). [Methods] Different explants from sterile seedling of Andrographis paniculata (Burm. f.) Nees were cultured in vitro to induce the production of fascicular buds. [ Results ] Cotyledon with nodes and half cotyledon with nodes were the most suitable explants for culture, and the budding rate was 100% and over 90% respectively. 6-Benzylaminopurine (BA) in the concentrations of 0.5 ~ 1.0mg/L were beneficial for the reproduction and growth of buds. The age of explants had no correlation with budding. The optimal rooting medium was MT (Murashige and Tucher) medium added with Img/L NAA (naphthaleneacetic acid) or IBA (indolebutyric acid). A highest rooting rate as much as 61.9% was achieved after 10 days of culture. [Conclusion] An effective plant regeneration system has been established for explants of Andrographis paniculata (Burm. f.) Nees.
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Objective To study the effects of density,phosphorous(P),and nitrogen(N)fertilizers on yield and quality of Andrographis paniculata.Methods Randomized block design in yield was used and HPLC was used to analyze the content of andrographolide and dehydroandrographolide.Results Plant height,fresh weight,dry weight,content of dehydroandrographolide were influenced significantly by density.The quality character and economic yield of content of andrographolide,dehydroandrographolide,total content of both andrographolide and dehydroandrographolide were significantly influenced by P nutrition.The economic yield of dry weight,content of andrographolide and dehydroandrographolide,and total content of both andrographolide and dehydroandrographolide were influenced significantly or extreme significantly by N nutrition.P and the interaction of P and N fertilizer extremely influenced the economic yield.The relative influences between plant weight and dry weight was extremely significance,that between quality character and quality character,that between quality character and economic yield were significant or extremely significant.Conclusion When planted with 1.8?105 /hm2,fertilizered with P2O5 540 kg/hm2 and pure N 225 kg/hm2,the hightest economic yield is gained.
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AIM: To study the HPLC fingerprints and establish a sensitive and specific method for the quality control of Andrographis paniculata(Burm.f.) Nees. METHODS: All 23 samples of Andrographis paniculata(Burm.f.) Nees collected from 10 different places were determined by RP-HPLC.Shimadzu-ODS column was used with maxtures of 0.1% formic acid-acetonitrile and 0.2% formic acid-water as mobile phase in gradient mode.The flow rate was set at 1.0 mL/min.The column temperature was set at 25℃ and detecting wavelength at 254 nm.Hierarchical clustering,nonlinear mapping and similarity criteria were applied to evaluate the fingerprints. RESULTS: The simple and specific method with good repeatability was established.There were different contents of each component contained in habitat samples produced in individual area. CONCLUSION: The proceeding is suitable to differentiate Andrographis paniculata(Burm.f.) Nees from different places conveniently and can be used for the quality control of this herb.