In silico antiplasmodial effects of phytocompounds derived from Andrographis paniculata on validated drug targets of different stages of Plasmodium falciparum

Background:Andrographis paniculata has been widely reported as an herbal plant for malaria treatment.The increasing rate of resistance to recommended antimalarial drugs has justified the need for a continuous search for new and more potent drugs that target all stages of the Plasmodium falciparum life cycle from natural plant sources.This study aimed to determine the antiplasmodial effect of phytocompounds derived from A.paniculata on the stages of plasmodium falciparum.Methods:Phytocompounds from A.paniculata were identified by Gas Chromatography-Mass Spectrophotometry(GCMS)analysis.The phytocompounds were screened for their druggability using Lipinski’s rule of five and subjected to Absorption,Distribution,Metabolism,Excretion,Toxicity(ADMET)and druglikeness analysis.The phytocompounds were docked against some validated drug targets at different stages of Plasmodium falciparum(hepatic,asexual,sexual,and vector targets)using PyRx software to analyze the inhibitory potential and protein-ligand interaction.Thereafter,the stability and flexibility of the best complexes were assessed through molecular dynamics simulations at 50ns using WebGRO.Result:The 7a-Isopropenyl-4,5-dimethyloctahydroinden-4-yl exhibited a higher binding affinity and better stability throughout the simulation period with P.falciparum dihydrofolate reductase-thymidylate synthase and Plasmodium falciparum M1 alanyl aminopeptidase for asexual blood stage and gametocyte stage of Plasmodium falciparum,respectively than the existing drugs.Meanwhile,N-Ethyl-3-methoxy-4-methylphenethylamine was also found to have a higher binding affinity and more stability throughout the simulation period with P.falciparum purine nucleoside phosphorylase and Plasmodium falciparum gametocyte surface protein for Hepatic schizonts stage of Plasmodium falciparum and gametocyte transmission blocking stage,respectively,than the existing drugs.Conclusion:The 7a-Isopropenyl-4,5-dimethyloctahydroinden-4-yl and N-Ethyl-3-methoxy-4 methylphenethylamine from A.paniculata are predicted as an antimalarial drug candidate.Thus,it is recommended that in vitro and in vivo bioassays be conducted on these hit compounds to validate these predictions.

Effects of Atmospheric Pressure Air Plasma Pretreatment on the Seed Germination and Early Growth of Andrographis paniculata

The objective of this paper is to demonstrate whether air plasma can change the seed germination characteristics, seedling emergence, as well as biochemical reactivity, in Andrographis paniculata （A. paniculata） seedlings by modifying the seed coat and finding a beneficial treatment dose. Eight treatment doses and one control were used to conduct electrical conductivity determination, a germination test, a seedling emergence test and a biochemical assay. The results showed that after being treated with air plasma excited at 5950 V for 10 s, the perme- ability of the seeds was improved significantly, resulting in the acceleration of seed germination and seedling emergence. In the meantime, the catalase activity and catalase isoenzyme expression were also improved, while the malondialdehyde content in the seedlings was decreased （which means greater counteraction with environmental stress）. After being treated with 4250 V for 10 s and 5950 V for 20 s, the seed germination was enhanced, but without an obvious change in seedUng emergence. However, after treatment with 3400 V for 20 s and 5100 V for 10 s, the permeability of the seeds was decreased, resulting in a delay in seedling emergence. These results indicate that air plasma can change the physiological and biochemical characteristics of Andrographis paniculata seeds by modifying the seed coat, combined with the effects of the active plasma species, and that different treating doses have different effects.

Mechanism of antagonistic effects of Andrographis paniculata methanolic extract against Staphylococcus aureus

Objective: To investigate the effects of Andrographis paniculata(Burm.f.) Wall. Ex Nees(A. paniculata) on expressions and activities of catalase, superoxide dismutase and alkylhydroperoxide reductase C in Staphylococcus aureus(S. aureus) with respect to its survival in vitro. Methods: Antioxidative property of methanolic leaves extract of A. paniculata(0.06 mg/m L). Minimum inhibitory concentration(MIC) was determined by its ability to reduce hydrogen peroxide(H_2O_2) toxicity against S. aureus ATCC 25923(3.8×10~8 cfu/m L). Effects of the extract on expressions of kat A(encoding catalase), sod A and sod M [encoding superoxide dismutases(SODs)], and ahp C [encoding alkylhydroperoxide reductase C(AhpC)] in S. aureuswere determined by RT-q PCR and corresponding enzyme activity assays were performed. Nitroblue tetrazolium reduction(NBT) assay was performed to determine effects of the extract on intracellular and extracellular levels of O_2-in S. aureus. Results: Cells challenged with 7.5 mmol/L H_2O_2 showed 0% survival in 30 min whereas 25% survived after treatment with the extract and H_2O_2. Cells that were treated with the extract alone had 43% survival in the same exposure period. Expressions of sod A and sod M genes in extract-treated cells were lowered 0.8-fold and 0.7-fold, respectively with decrease in total SOD activity of 26.8 U compared to untreated cells, 32.4 U(P<0.05). In contrast, extract-treated S. aureus cells showed 3.3-fold increase in kat A expression with corresponding increase in catalase activity of 1.828 U compared to untreated cells which was 1.248 U,(P<0.05). More profoundly, ahp C expression was increased 61-fold in extract-treated cells,(P<0.05) with corresponding increase in Ahp C activity of 0.018 U compared to untreated cells, 0.012U,(P<0.05). Extract-treated cells had significantly lower intra-and extracellular O_2-levels with absorbance readings(A_(575nm)) of 0.340 and 0.524 compared to untreated cells which were 0.516 and 0.928(P<0.05), respectively. Conclusions: Taken together these results suggest that the low MIC of A. paniculatamethanolic leaves extract(0.06 mg/m L) reduce H_2O_2 toxicity and more importantly, was in itself effectively inhibitory against S. aureus. Further, our observations suggest that a probable mode of its inhibitory mechanism against S. aureus is by reducing total SOD activity through downregulation of sod A and sod M expressions.

A new ent-labdane diterpenoid lactone from Andrographis paniculata

A new ent-labdane diterpenoid lactone was isolated from Andrographis paniculata.Its structure was identified on the basis of spectral data including 2D NMR.

A new ent-labdane eiterpenoid lactone from Andrographis paniculata

A new ent-labdane diterpenoid lactone with a new natural product was isolated from Andrographis paniculata. Their structures were elucidated on the basis of spectral evidences including 2D NMR.

Antidiabetic effect of combination of fractionated-extracts of Andrographis paniculata and Centella asiatica: In vitro study

Objective: To examine the effect of combination of Andrographis paniculata herb fraction(AHF) and Centella asiatica herb fraction(CHF) on PPAR γ and GLUT4 mRNA expression in 3 T3-L1 adipocyte, and its effect on insulin-stimulated glucose uptake. Methods: 3 T3 L1 adipocyte cells were used to investigate gene expression of PPARy reverse transcription-polymerase chain reaction method. The γ and GLUT4 protein b adipocyte cells were differentiated by using insulin, dexamethasone and 3-isobutyl-1-methylxanthine from 3 T3-L1 cells. Pioglitazone, AHF, CHF and the combination of both herbs were evaluated on glucose uptake activity, PPAR γ and GLUT4 mRNA expressions in 3 T3-L1 adipocyte. Results: The results showed that combination of AHF at 30 μg/mL and CHF at 10 μg/mL could enhance insulin-stimulated glucose uptake. The combination also increased PPAR γ and GLUT4 mR NA expressions significantly in comparison to those of negative control(DMSO). These effects were equal in comparison to those of pioglitazone(0.02 μM) and its single extracts Conclusions: The combination of AHF and CHF can increase glucose uptake and insulin sensitivity through up-regulation of PPAR γ and GLUT4 mRNA expressions in 3 T3-L1 adipocyte.

Metabolic disturbance and phytochemical changes in Andrographis paniculata and possible action mode of andrographolide

Objecive: To explore the effect of gibberellic acid(GA_3) and its inhibitor paclobutrazol(PBZ)on chemical composition and their pharmacological effects on Andrographis paniculata(Burm.f.) Wall. ex Nees, and to clarify action mode of andrographolide.Methods: The chemical composition was extracted by sequential extraction with hexane, dichloromethane, ethyl acetate and methanol, respectively. Andrographolide and its derivatives were evaluated by HPLC. Moreover, the metabolic profiling was analyzed by GC-MS. Inhibitory effect of crude extracts was tested against Staphylococcus aureus using agar well diffusion method. Mode of action was tested against mutant yeast by spotting assay. Andrographolide were tested for their mode of action against eukaryotes. Rsults: Among different solvents, dichloromethane gave the highest yield of crude(3.58% DW), with the highest andrographolide content(8.3 mg/g DW). The effect of plant hormone(10 mg/L GA_3 or PBZ) on phytochemical variations and bioactivity of Andrographis paniculata was demonstrated. It was found that PBZ promoted sesquiterpene compounds about 3.5 times over than GA_3 treatment. But inhibitory effect of extracts against Staphylococcus aureus was highest in GA_3 treated plants; andrographolide and 14-deoxy-11,12-didehydroandrographolide contents were significantly higher than those of water or PBZ. It was found that there were 11 strains involving in ergosterol biosynthesis, V-ATPase activity and homeostasis, and superoxide detoxification process. In this regard, andrographolide might cause the damage on the lipid bilayer of yeast cell and plasma membrane by interfering ergosterol biosynthesis.Conclusions: It is found that GA_3 promotes andrographolide and 14-deoxy-11,12-didehydroandrographolide content while PBZ promotes sesquiterpene content. Andrographolide might cause the damage on the lipid bilayer of yeast cell and plasma membrane by interfering ergosterol biosynthesis. It might also affect mitochondria electron transport chain, leading to the occurrence of ROS, which can further harm cell organelles. However, the library screening is the first step to investigate mode of action of andrographolide.

Comparison study of anti-microbial activity between crude extract of Kappaphycus alvarezii and Andrographis paniculata

Objective: To evaluate the antimicrobial properties of Kappaphycus alvarezii(K. alvarezii) and Andrographis paniculata(A. paniculata) and to compare the microbial inhibition activities between these two crude extracts.Methods: Both K. alvarezii and A. paniculata were extracted with methanol before the commencement of antimicrobial properties studies. There were a total of eight species of bacteria, including Gram-positive bacteria Bacillus cereus, Bacillus subtilis, Staphylococcus aureus and Staphylococcus epidermidis and Gram-negative bacteria, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Salmonella enterica. The antimicrobial activity was tested by disk diffusion method.Results: Crude extract of K. alvarezii was found not effective against both Grampositive and Gram-negative bacteria. In contrast, A. paniculata showed higher inhibition towards the growth of Gram-positive bacteria compared to Gram-negative bacteria.Results revealed that Bacillus subtilis was susceptible at lower concentration of A. paniculata crude extract however Staphylococcus epidermidis was the most susceptible towards A. paniculata at higher concentration. Although the inhibition zones produced by the crude extract were smaller than that of the positive control, streptomycin disc. A. paniculata crude extract still can be considered as potential antimicrobial agents either because it is a natural product or the active compound which is yet identified from its crude extract.Conclusions: Crude extract of K. alvarezii has zero inhibition in bacteria growth whereas A. paniculata exerted higher inhibition towards Gram-positive bacteria. The bioactive compounds contained by A. paniculata can be evaluated in order to yield a better vision towards the mode of action.

Harvesting Age and Time Effect of Andrographis paniculata Leaf on Its Anti-Inflammatory Activity

In this study, Fourier Transform Infrared (FTIR)-, 1H Nuclear Magnetic Resonance (NMR)-spectroscopy and multivariate statistical analysis were used for the screening of in vitro anti-inflammatory activity on three different germplasm accessions namely 11,341 (P), 11,248 (T) and 11,265 (H) of Malaysian Andrographis paniculata (A. paniculata) leaf. The anti-inflammatory activity by nitric oxide (NO) inhibition assay in relation to the different harvesting ages and times of A. paniculata leaf was determined through comparison by partial least-squares analysis (PLS) using SIMCA-P. The PLS allowed the separation and correlation between the NO assay with the phytochemical present due to the effects of different harvesting ages and times. From the PLS plots, conclusions were drawn with respect to the correlation between A. paniculata leaf metabolites with the anti-inflammatory results which showed that 180 days after transplanting (DAT) of morning session for accessions T and H, and evening for P gave the highest anti-inflammatory activity.

The Effect of Andrographis Paniculata Nees(APN)in Alleviating the Myocardial

In experimental dogs,the effect of APN in alleviating the ischemiareperfusion injury was prominent.Compared with the sustained isthmian group,superoxide dismutast(SOD)in the ischemic region of myocardial tissue in the ischemiareperfusion group was significantly decreased and malondialdehyde(MDA)marktdly increased Ca2+in myocardial cells was increased. and ultrastructural changes of myocardial tisstes were severe in the APN-pretreated ischemia-reperfusion group ,on the contrary,all the above parameters showed reversely,i,e.,SOD increased ,MDA and intracellular Ca2+ decreased.the ultrastructure changes were less distorted.

Two new diterpenoid lactones isolated from Andrographis paniculata

In the present study, two new diterpenoid lactones, 3-deoxy-andrographoside(1) and 14-deoxy-15-methoxy-andrographolide(2), were isolated from the aerial parts of Andrographis paniculata. Their structures were elucidated by combination of NMR, MS, and chemical methods. The configurations of 1 and 2 were established based on the analysis of ROESY data and single crystal X-ray diffraction experiment.

New Diterpenoids from Andrographis paniculata （Burm. f.） Nees

To investigate diterpenoids from the aerial parts of Andrographis paniculata （Burm. f.） Nees, three new ent-labdane diterpenoids, namely 19-norandrographolides A-C （compounds 1-3）, were isolated from the ethanolic extract of A. paniculata. Their structures were established by HRESIMS and NMR spectral data in combination with X-ray crystallographic analysis.

A new biocompatible microemulsion increases extraction yield and bioavailability of Andrographis paniculata

The purpose of this study was to design and prepare a biocompatible microemulsion of Andrographis paniculata(BMAP) containing both fat-soluble and water-soluble constituents. We determined the contents of active constituents of BMAP and evaluated its bioavailability. The biocompatible microemulsion(BM), containing lecithin and bile salts, was optimized in the present study, showing a good physical stability. The mean droplet size was 19.12 nm, and the average polydispersity index(PDI) was 0.153. The contents of andrographolide and dehydroandrographolide in BMAP, as determined by high performance liquid chromatography(HPLC), were higher than that in ethanol extraction. The pharmacokinetic results of BMAP showed that the AUC0-7 and AUC0→∞ values of BMAP were 2.267 and 27.156 μg·m L-1·h 1, respectively, and were about 1.41-fold and 6.30-fold greater than that of ethanol extraction, respectively. These results demonstrated that the bioavailability of and rographolide extracted by BMAP was significantly higher than that extracted by ethanol. In conclusion, the BMAP preparation displayed ann improved dose form for future clinical applications.

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 Andrographispaniculata. 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.

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.

ABA and GA_(3)affect the growth and pigment composition in Andrographis paniculata Wall.ex Nees.,an important folk herb

In this study,5μmol·L^(-1)abscisic acid(ABA)and gibberellic acid(GA3)were used to study the effect of both growth regulators on the morphological parameters and pigment composition of Andrographis paniculata.The growth regulators were applied by means of foliar spray during morning hours.ABA treatment inhibited the growth of the stem and internodal length when compared with control,whereas GA3 treatment increased the plant height and internodal length.The total number of leaves per plant decreased in the ABA-treated plants,but GA3 treatment increased the total number of leaves when compared with the control.Both growth regulators(ABA and GA3)showed increased leaf area.ABA and GA3 treatments slightly decreased the total root growth at all the stages of growth.The growth regulator treatments increased the whole plant fresh and dry weight at all stages of growth.ABA enhanced the fresh and dry weight to a larger extent when compared with GA3.An increase in the total chlorophyll content was recorded in ABA and GA3 treatments.The chlorophyll-a,chlorophyll-b,and carotenoids were increased by ABA and GA3 treatments when compared with the control plants.The xanthophylls and anthocyanin content were increased with ABA and GA3 treatments in A.paniculata plants.

Assessing suitability of Andrographis paniculata genotypes for rain-fed conditions in semi-arid climates

Medicinal plants are generally suggested for degraded land or areas having low rain-fall.Andrographis paniculata is an important medicinal plant known for its diterpene lactone i.e.andrographolide.We exposed 26 genotypes of A.paniculata to rain-fed condition during early-season rain-fed(ESRF)and mid-season rain-fed(MSRF)conditions and compared with the control to assess their suitability in semi-arid climate region of Gujarat,India.The study was performed during two consecutive years 2016–2017 in split plot design with rain-fed conditions as main-plot treatment and genotypes as sub-plot treatment in three replications.The gaseous exchange parameters taken at 85100 days after transplanting exhibited mean photosynthesis rate was 20.5 lmol CO_(2) m^(-2)s^(-1) ranging between 15.9 lmol CO_(2) m^(-2)s^(-1) and 24.0 lmol CO_(2) m^(-2)s^(-1).Exceeding 7%decrease in stomatal conductance was recorded compared to the control plants under imposed rain-fed condition.The mean value of transpiration rate was 4.8 mmol H2O m^(-2)s^(-1).The mean value of water use efficiency was 4.6.MSRF condition decreased leaf water potential from-0.517 to-1.189 Mpa.The dry herbage decreased by 29.2%due to MSRF condition as compared the control.The andrographolide yield per plant suffered significantly and under ESRF condition it reduced mainly due to reduction in andrographolide content.Reduced andrographolide yield per plant in MSRF condition was mainly due to reduction in herbage yield.Results revealed that ESRF and MSRF condition characterized by erratic rainfall pattern and dry spell is not beneficial for quality production in A.paniculata.Based on the least decrease in andrographolide,yield two genotypes viz.,AP 6 and AP 12 identified to be suitable for ESRF condition and two genotypes viz.,AP 19 and AP 6 suitable for MSRF condition.Overall,four genotypes,AP 35,AP 39,AP 61 and AP 24 were suitable as the promising genotypes having high andrographolide yield.

An Experimental Study of the Mechanism of Andrographis Paniculata Nees（APN） in Alleviating theCa^（2＋）－overloading In the Proc

The aim of this experiment is to study the mechanism of APN in alleviating the Ca2+-overloading in dog model during the process of ischemic reperfusion. In comparison with the sustained ischemic group, the parameters in the ischemic reperfusion group demonstrated: Ca2+ of ischemic region of myocardial cell increased (P<0.05) , Na+ increased remarkably(P<0. 01), the activity of Ca2+-ATPase dropped remarkably(P<0. 01),and MDA increased significantly(P<0. 01). Whereas in the group pretreated with APN, the Ca2+ in the relevant area reduced (P<0.05), Na+ decreased significantly(P<0. 01), the activity of Ca2+-ATPase and Na+-K+ ATPase increased remarkably(P<0.01) ,and MDA decreased significantly (P<0. 01). These findings indicate that APN may improve the activity of sarcolemma ATPase in alleviating the Ca2+ and Na+-overloading by decreasing the harmful effect of oxygen free radicals.

穿心莲种质产量性状与药用成分分析

穿心莲(Andrographis paniculata(Burm.f.)Nees)是我国传统的中药材,应用广泛。为了评价穿心莲种质的遗传多样性,收集了广东湛江和广西贵港两个主要产区的种质材料,对其产量性状和主要药用成分进行调查和分析。结果表明,同一来源穿心莲材料的产量性状及药用成分均呈现较大变异。产量性状中,单株生物量变幅最大为100~624 g,其余从大到小依次为叶片质量、叶面积、一次分枝数、生育期、株高。药用成分性状中,穿心莲内酯变幅最大为1.5385~6.5944 mg/g,其余依次为新穿心莲内酯、14-去氧穿心莲内酯、脱水穿心莲内酯。供试材料分析发现种质纯和度差,具有显著的遗传多样性,为材料鉴选及遗传育种提供了丰富的材料基础。此外,相关性分析表明,穿心莲主要产量性状与主要药用成分含量间呈显著负相关。材料选育时需要注重产量性状和药用成分之间的平衡。最后,进一步按产量性状及药用成分指标进行聚类分析,筛选重要材料。本研究结果可为穿心莲的种质资源保护及良种选育提供育种研究材料基础。

穿心莲对斑点叉尾鮰消化酶活性的影响

为研究穿心莲(Andrographis paniculata)对斑点叉尾鮰(Ictalurus punctatus)消化酶活性的影响,试验选取平均体重为(5.92±0.63)g的健康斑点叉尾鮰270尾,随机分成9组(G0、G1、G2、G3、G4、G5、G6、G7、G8组),每组设3个重复,每个重复10尾鱼,每组分别投喂添加不同比例穿心莲粉(0%、0.38%、0.75%、1.13%、1.50%、1.88%、2.25%、2.63%、3.00%)的饲料。饲养试验时间为30 d。结果发现,肠道中的消化酶,G0和G1组的脂肪酶活性显著高于其它各组(P<0.05);G8组的淀粉酶活性最高,显著高于G0、G1、G4和G7组(P<0.05);胰蛋白酶活性总体呈现出先上升后下降的趋势,以G6组的最高。结果表明,在斑点叉尾鮰饲料中添加1.88%~2.25%的穿心莲能提升胰蛋白酶的活性。