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Background: Crateva unilocularis, Millettia extensa, and Plumbago zeylanica are commonly used traditional medicinal plants in Nepal for treating various skin ailments. Objectives: The ethanol extracts of these plants were analyzed for qualitative and quantitative phytochemical content, antioxidant potential, and anti-tyrosinase activity. Methods: The Folin-Ciocalteu technique was used to measure the total phenolic content, and the aluminum chloride method was used to assess flavonoid content.In vitro antioxidant activity was evaluated via 2,2- diphenyl-1-picrylhyrazyl (DPPH), nitric oxide (NO), and hydrogen peroxide (H2O2) assays. A mushroom tyrosinase inhibition assay was employed to assess anti-tyrosinase activity. Molecular docking was performed using the tyrosinase enzyme (PDB ID: 2Y9X) for the five selected phytoconstituents of Millettia extensa. Results: The ethanolic extract of Millettia extensa leaves exhibited high concentrations of phenolic compound (273.85 ± 4.05 mg GAE/g dry extract weight) and flavonoids compound (1787.35 ± 0.22 mg QE/g dry extract weight). The roots of Plumbago zeylanica showed the highest levels of DPPH and NO radical-scavenging activities, with percentages of 95.83% and 43.83%, respectively. Similarly, the stem bark of Crateva unilocularis showed notable H2O2 radical scavenging activity (67.81%). Millettia extensa root bark demonstrated the most potent tyrosinase inhibition activity (99.31 ± 0.16), comparable to the standard drug Kojic acid (99.56 ± 0.08). Molecular docking corroborates the in vitro anti-tyrosinase activity, with isoauriculatin and auriculatin demonstrating the highest binding affinities of −9.9 and −9.7 kcal/mol, respectively, compared to Kojic acid (−5.6 kcal/mol). Conclusion: These findings highlight the significant tyrosinase inhibitory potential of Millettia extensa root bark, suggesting further exploration of its cosmetic application and safety evaluation. [ABSTRACT FROM AUTHOR]

AbstractThe study utilizes an analytical quality-by-design (AQbD) approach to establish a robust and efficient HPTLC method for plumbagin quantification in plumbago species. The highest-risk critical method parameters (CMP; concentration of plumbago and composition of mobile phase) and most appropriate critical analytical attributes (CAA; Peak area and retention factor [Rf]) were employed in the AQbD methodology. These CMP were tuned for higher plumbagin recovery using the central composite design (CCD) experiment. The created approach was validated by the requirements outlined in the International Conference on Harmonization (ICH) Q2 (R2). Every plate involved in the proposed method validation has been subjected to the passing requirements of the system suitability test (SST). The low LOD (16.93 ng/mL) and LOQ (49.68 ng/mL) values and the minimal divergence in intra- and inter-day studies validate the developed method’s high precision and sensitivity. Further experiments employing the proposed method to determine the amount of plumbagin from the two distinct plant sources harvested under two different climate conditions determined the highest plumbagin content in P. indica, which is in line with the results. To summarize, the AQbD method may be used in other plant systems to optimize various factors and get a greater concentration of the desired plant metabolite. [ABSTRACT FROM AUTHOR]

This study aimed to develop and evaluate a niosomal formulation loaded with Plumbago zeylanica extract (PZE). The solvent evaporation method was used to prepare niosomes and optimized using the Box–Behnken design. The optimized formulation (PZE-Ns-Opt) was characterized for drug release, DPPH assay, α-amylase inhibition assay, α-glucosidase inhibition assay, and confocal laser scanning microscopy (CLSM) study. PZE-Ns-Opt showed a vesicle size of 253.6 nm, PDI of 0.108, entrapment efficiency of 62.4%, and drug release of 84.01%. The CLSM image of the rat's intestine suggested that the rhodamine red B-loaded PZE-Ns-Opts showed superior penetration compared to the control. Further, the antioxidant activity of the prepared formulation was exhibited as 89.46%±0.016 as compared to PZE (78.10%±0.005). In addition to this, α- amylase activity was inhibited by 95.11%±4.62, 85.88%±2.56, and 89.87%±3.65 by acarbose, PZE, and PZE-Ns-Opt, respectively. In comparison, the α-glucosidase activity was inhibited by 88.47%±1.04, 81.07%±0.50 and 85.78%±0.71 by acarbose, PZE, and PZE-Ns-Opt, respectively. To conclude, the development of PZE-Ns-Opt formulation and its characterization showed the establishment's authenticity. The in vitro antidiabetic, antioxidant studies expressed the protective effect from oxidative stress, CLSM of rat's intestine ability to penetrate to great extent and could be a promising candidate for the management of diabetes mellitus. [ABSTRACT FROM AUTHOR]

Simple Summary: Diabetic retinopathy is a severe complication of diabetes that can lead to vision loss due to the abnormal growth of blood vessels in the retina. This study investigates how two traditional medicinal plants, Solanum xanthocarpum and Plumbago zeylanica, might be used to treat this condition. We focused on identifying the active compounds in these plants and understanding how they interact with proteins related to diabetic retinopathy. Using advanced computer simulations, we found that these plant compounds effectively bind to and influence key proteins involved in the progression of the disease. Specifically, our research highlighted that compounds from Solanum xanthocarpum and Plumbago zeylanica could target several crucial pathways and proteins associated with diabetic retinopathy. These findings suggest that these plant-derived compounds have significant potential as therapeutic agents for diabetic retinopathy. By offering a new approach to treatment, this research could help improve the vision and overall quality of life of individuals suffering from this challenging condition. (1) Background: Diabetic retinopathy (DR) is a major complication of diabetes, marked by abnormal angiogenesis, microaneurysms, and retinal hemorrhages. Traditional Ayurvedic medicine advocates multi-target strategies for DR management. However, the mechanisms by which Solanum xanthocarpum (SX) and Plumbago zeylanica (PZ) exert therapeutic effects are not well understood; (2) Methods: To investigate these mechanisms, we employed network pharmacology and molecular docking techniques. Phytochemicals from SX and PZ were identified using the IMPPAT database and Swiss Target Prediction tool. DR-related protein targets were sourced from the GeneCards database, and common targets were identified through Venn diagram analysis. STRING and Cytoscape were used to construct and analyze protein–protein interaction networks. Pathway enrichment was performed with Gene Ontology and KEGG databases; (3) Results: We identified 28 active phytoconstituents, targeting proteins such as EGFR, SRC, STAT3, AKT1, and HSP90AA1. Molecular docking and dynamics simulations confirmed the strong binding affinities of these compounds to their targets; (4) Conclusions: The study highlights the multi-target activity of SX and PZ, particularly in pathways related to EGFR tyrosine kinase inhibitor resistance and PI3K–AKT signaling. These findings provide valuable insights into their therapeutic potential for DR, suggesting the effective modulation of key molecular pathways involved in the disease. [ABSTRACT FROM AUTHOR]

Objective: The concurrent usage of oral hypoglycemic agents produces side effects, and the herbal medicine Plumbago zeylanica L. (PZ) is being studied to reduce these effects. The antioxidant and antidiabetic effects of PZ roots on streptozotocin-nicotinamide-induced diabetic rats were investigated. Materials and Methods: Triple maceration method was used for the development of PZ extracts. Standardization of the plant extract and in vitro study was carried out by the physicochemical study, 1, 1-diphenyl-2-picryl-hydrazyl, and α-amylase methods. Animals were divided into seven groups, and the experiment was designed into two parts. Seven groups were taken normal control, diabetic control, hydroalcoholic extract of PZ at different doses (100–200 mg/kg), and metformin (MET, 25 mg/kg) for 21 days. The coadministration of a low dose of PZ (100 mg/kg) and MET (100 mg/kg) was administered orally. Blood glucose level (BGL) was measured continuously for 3 weeks of overnight-fasted animals. Biochemical estimation was assessed by total protein, catalase, nitric oxide, and reduced glutathione content. Histological features of the β-cells were observed through histopathological study. Results: Hydroalcoholic PZ (HAPZ) showed the highest antioxidant and antidiabetic activities, respectively. Oral administration of HAPZ extract (200 mg/ml) for 21 days diminished the BGL significantly from 298 ± 7.05 to 230 ± 5.69 mg/dL, whereas coadministration of HAPZ (100 mg/kg) with MET (10 mg/kg) also reduced BGL from 286 ± 4.10 to 231 ± 5.75 mg/dL. The best outcomes were obtained when PZ extract (100 mg/kg) was combined with a low dose of MET (10 mg/kg). Conclusions: We can conclude that PZ might be the alternative to synthetic medicines for the management of diabetic disorders.

Metabolite stuffing may depend upon the session of the collection, soil condition, and geographical regions of the collection site, and this impacts the quantitative production and biological potential of the plant species. A total of 14 accessions of Plumbago zeylanica were collected from different locations in Central India to cover the entire range of topography from the foothills up to the highest peak. The instrumental thin-layer chromatography‒ultraviolet detection (instrumental TLC‒UV) method was used to quantify plumbagin and unweighted pair group method with arithmetic mean (UPGMA) cluster analysis to identify elite sources. The present investigation aims to identify the best quality planting material of P. zeylanica, in terms of elite chemotypes collected from different locations of Central India. UPGMA cluster analysis revealed the variation of the plumbagin content (0.0038‒0.6091% dry weight basis) among the collected accessions of P. zeylanica. The cluster analysis of plumbagin content depicted the distinctness in plant metabolite contents between the individuals as they were clustered by difference between accessions. This study will also promote the use of P. zeylanica in herbal drug development and will aid in the site-specific exploration of elite chemotype(s) with validated pharmacological action to meet the medicinal and commercial demands. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica L., a wild shrub, is a vital natural source of plumbagin, a potent 1,4-naphthoquinone renowned for its anti-cancer properties, notably effective against breast, prostate, and ovarian cancers. Traditional plumbagin extraction, involving root uprooting and plant destruction, raises ecological concerns. The primary objective of this study is to enhance plumbagin production by incorporating the elicitation process into in vitro cultivation with regenerated plants that retain all of their intact organs. Seven different elicitors categorized into three distinct groups were employed to stimulate plumbagin content. Among the various elicitors used, this study marks the first application of biogenic silver nanoparticles (AgNPs) from Curcuma amada in stimulating plumbagin production in this plant. The maximum plumbagin content, recorded at 8.98 ± 0.24 mg/g dry weight basis, was found in the roots when elicited with AgNPs at a concentration of 15 mg/l. In addition to that, biotic elicitors (yeast extract, chitosan and casein hydrolysate) and heavy metals (lead, cobalt and nickel) also successfully elicit plumbagin in the root and aerial parts of the plants, quantified through High Performance Liquid Chromatography (HPLC). In our study, we found that certain elicitors induced root browning and tissue necrosis, as confirmed by propidium iodide (PI) staining. The most significant browning effects were observed with chitosan from biotic sources and lead from heavy metals, while no such effects were associated with AgNPs at any concentration. Utilizing intact, entire plants as the subjects for elicitation in our study is a valuable aspect. This approach closely replicates the natural process occurring in intact plants, enhancing the relevance of our findings to practical situations.Key message: Utilizing silver nanoparticles alongside diverse biotic and heavy metal elicitors applied to entire plants in liquid cultures facilitates the sustainable production of plumbagin from various plant organs. [ABSTRACT FROM AUTHOR]

Objectives: We investigated the anticancer activity of the PzMH fraction (hexane fraction) extracted from the roots of Plumbago zeylanica L., an ethnomedicinally significant plant widely distributed in India. Methods: The PzMH fraction was obtained through rigorous extraction and purification processes. In vitro cytotoxicity assays were performed to assess its effects on Ehrlich ascites carcinoma (EAC) cells. Acute toxicity studies were conducted to evaluate its safety profile. An in vivo study was carried out on EAC-bearing Swiss albino mice to assess its anticancer efficacy. Flow cytometric and microscopic analyses were done to examine the induction of cell death by the PzMH fraction. Western blot analysis was used to investigate the molecular mechanisms involved. Results: The PzMH fraction exhibited a significant cytotoxic effect in vitro, resulting in 50% cell death in EAC cells at low concentrations. The calculated GI50 value for the PzMH fraction was 42.74 µg/ml, demonstrating a comparable efficacy to the standard drug 5-fluoro uracil (GI50 = 43.38 µg/ml). The safety of therapeutic doses was confirmed through acute toxicity studies, which yielded an LD50 value of 500 mg/kg body weight. In the in vivo study, the PzMH fraction demonstrated a substantial 79.05% inhibition in the growth of EAC cells at the 300 mg/kg body weight dose of PzMH fraction. Flow cytometric and microscopic analyses revealed distinct apoptotic features in EAC cells treated with the PzMH fraction. Cell cycle analysis showed a significant arrest at the G0/G1 stage following treatment. PzMH treatment elicited a response characterized by escalated levels of cleaved caspases-3 and −9, while concurrently leading to a decreased expression of the Bcl2 protein, as evidenced by Western blot analysis. Conclusions: The current research provides empirical evidence supporting the anticancer activity of the PzMH fraction extracted from P. zeylanica. The observed cytotoxicity, safety, and apoptosis-inducing properties make it a promising candidate for further investigation as a potential cancer therapy. Further exploration of its phytochemical composition, including major compounds such as 4-hydroxybenzaldehyde, trans-cinnamic acid, plumbagin and lawsone, contributes to our understanding of its mechanisms of action.

Plumbago zeylanica has been used in the traditional system of medicine from thousands of years owing to its potential therapeutic properties, however the anticancer and anti-metastatic effects are largely unknown against cervical cancer cells. In this study we demonstrated the cytotoxicity activity, inhibition of cell migration, induction of S-phase cell cycle arrest and the down regulation of expression level of epidermal growth factor receptor (EGFR) by ethanol extract of the root of P. zeylanica (ERPZ). The cytotoxicity effects were analyzed by MTT assay the IC50 was determined to be 10.49 ​μg/ml after 72 ​h of incubation. Besides, strong inhibition of wound healing activity was observed at 20 ​μg/ml concentration, where the wound size was reduced to 33%. The ethanol root extract of P. zeylanica also showed significant S-phase cell cycle arrest of 68.9% at 20 ​μg/ml concentration in HeLa cells which was accompanied by the down regulation of EGFR. Ethanol extracts of the root of P. zeylanica (ERPZ) treatment inhibited the growth of cervical cancer cells. The ERPZ arrested cells at the S-phase of the cell cycle. Besides the HRLC-MS analysis of ERPZ identified 15 compounds and among which the five major compounds such as 7,8-dihydroxy-4-methylcoumarin, neodiosmin, diosmetin, hispidulin, and formononetin were reported to possess antioxidant and anticancer activities, could plausibly induce cell death in HeLa cells.

The present study describes a simple, reproducible, and cost-effective regeneration system using seaweed liquid extract (SLE) of Sargassum ilicifolium supplemented in Murashige and Skoog (MS) medium for in vitro seed germination, multiple shoot and root induction, and plumbagin accumulation in P. zeylanica. High rate of seed germination and seedling length was recorded on ½ − MS + SLE (2%) which was 1.11- and 1.28-fold higher as compared to GA3 (2.89 µM). The best response in terms of explants producing maximum number of shoots was recorded on MS medium supplemented with 4% SLE that was 2.26- and 1.37-fold higher than BAP (3.33 µM) alone and in combination with IAA (4.28 µM), respectively. High frequency root induction and plumbagin accumulation was recorded on ½ − MS + 2.5% SLE which showed 1.34-, 1.49-, and 1.44-fold increase for the roots per shoots, average root length, and plumbagin, respectively when compared with IBA (6.15 µM). Besides being an alternative to commercially available PGRs, this is the first report demonstrating the potential of SLE for enhanced growth and plumbagin accumulation in P. zeylanica. Key message: Efficient in vitro mass propagation protocol of P. zeylanica is developed using S. ilicifolium SLEs. SLEs with MS media were a substitute to the commercially available PGRs for micro-propagation and plumbagin production. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica shows the presence of many phytochemical constituents of which plumbagin, a type of naphthoquinone, is vital and has major biological effects. Plumbagin is majorly present in the root of the plants, with a minimal concentration in other parts of the plant. However, extracting plumbagin from the roots is destructive harvesting. Thus, micropropagation could be a viable alternative to reduce the collection of plant from wild. Study from micropropagated leaf calli was considered as a steady and sustainable source of phytochemical. P. zeyalnica callus was subcultured in an appropriate medium with plant growth regulators to initiate root cultures. A comparative study was performed to estimate the quantities of plumbagin obtain from the wild plant root part and root created by callus culture differentiation. [ABSTRACT FROM AUTHOR]

There has been tremendous spread of antimicrobial resistance globally, mainly due to the excessive and unnecessary use of antibiotics, making the situation alarming. This has created a need for the development of alternative strategies to selectively target the bacterial pathogenicity without exerting selection pressure for the development of antimicrobial resistance. Targeting quorum sensing (QS)-mediated virulence and biofilms by nontoxic natural products is gaining importance as new control strategy to combat the virulence and biofilms of pathogenic bacteria. In this study, the crude extract of Plumbago zeylanica was fractioned in different solvents using liquid-liquid partitioning to obtain the most bioactive fraction. The inhibitory effect of the bioactive extract of P. zeylanica on QS at sub-minimum inhibitory concentrations (MICs) was studied against Chromobacterium violaceum 12472, Pseudomonas aeruginosa PAO1, and Serratia marcescens MTCC 97. Biofilm inhibition was studied using microtiter plate assay, scanning electron microscopy, and confocal laser scanning microscopy. Major phytocompounds detected were cinnamaldehyde dimethyl acetal, plumbagin, asarone, 4-chromanol, phthalic acid, palmitic acid, ergost-5-en-3-ol, stigmasterol, and ß-sitosterol. The violacein production in C. violaceum 12472 was reduced by >80% in the presence of P. zeylanica hexane fraction (PZHF; 200 µg/ml). The most active PZHF inhibited QS-mediated virulence factors of P. aeruginosa PAO1 such as pyocyanin, pyoverdin, rhamnolipid production, motility, etc., significantly at sub-MICs. Similarly, PZHF showed 59 to 76% inhibition of biofilm formation of above test pathogens. The findings revealed that active fraction of P. zeylanica was effective against the QS-regulated functions and biofilms development of Gram -ve pathogenic bacteria. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica L., commonly known as leadwort or Ki Encok plant in Indonesia, has some beneficial effects on human health, such as anticancer, anti-microbial, anti-inflammatory, wound healing, and anti-diabetic. Therefore, this experimental study aims to evaluate the chloroform extracts of P. zeylanica L. leaves for the isolation of the phytoconstituents. The crushed form of the leaves from South Sumatera (6.5 g) was macerated in 65 ml of chloroform for 5 days, while a total of 80 g leaves powder from Central Java was dissolved in 800 ml of chloroform. The extracts were filtered and evaporated to dryness using a rotary evaporator. Subsequently, the identification of secondary metabolites was carried out using Gas Chromatography-Mass Spectrometry (GC-MS). In conclusion, the leaves of P. zeylanica L. contain β -sitosterol, lupeol, plumbagin, phytol, and lauric acid. [ABSTRACT FROM AUTHOR]

Abstract Background Plumbagin is one of the pharmaceutically important biomolecule with anticancer potential. Among the plants reported to produce plumbagin, P. zeylanica topped the list. The plumbagin production is very slow with low yield and maximum 0.5% (of dry weight) was reported in P. zeylanica. To meet the increasing demand of the plumbagin at global level, the P. zeylanica are exploited at commercial level, which may pose serious threat on the germplasm of the plant populations. So, it is needed to enhance the contents of plumbagin in P. zeylanica using biotechnological approaches. Among the various methods used to enhance the contents of plumbagin in P. zeylanica, utilization of fungal endophytes to enhance the plumbagin contents is a widely accepted approach. As fungal endophytes have the potential to synthesize various secondary metabolites and also reported to influence the synthesis of the secondary metabolites in plants. In the present study, an attempt was made to assess the effect of fungal endophytes of the Plumbago zeylanica L. on enhancement of plumbagin contents at in vivo level. Results Total 3 fungal endophytes were recorded from the roots of P. zeylanica collected from Khadki, Pune. The fungal endophytes were identified at morphological and molecular level. After 1 year of the treatment with fungal endophytes, significant enhancement of plumbagin was recorded in the roots of the P. zeylanica. Plumbagin contents in each were quantified against the standard plumbagin by employing LCMS-MS technique. Among the three fungal endophytes, the maximum enhancement of plumbagin content (122.67%) was reported with the treatment of Alternaria sp. (Isolate-3) in the roots of the P. zeylanica compared to control. Conclusion Among the three fungal endophytes, the maximum enhancement of plumbagin content (122.67%) was reported with Alternaria sp. (Isolate 3) in the roots of the pot-grown plants of P. zeylanica at in vivo level.

A high-performance thin-layer chromatography (HPTLC) method has been developed for the quantitative determination and validation of plumbagin in the methanolic fraction of Plumbago zeylanica. For achieving good separation, the mobile phase of toluene‒ethyl acetate (9:2, V/V) for plumbagin was used. Densitometric determination of plumbagin was carried out at 270 nm in reflection/absorption mode; the calibration curves were linear in the range of 100‒600 ng/spot. During analysis, the methanolic fraction of P. zeylanica L. showed the presence of plumbagin. The method is specific, simple, precise, and accurate. The obtained data can be used for the routine analysis of the reported biomarkers in crude drugs and extracts. The quantification and the method validation of these biomarkers have not yet been reported in P. zeylanica which can be utilized for the proper standardization of the plant. [ABSTRACT FROM AUTHOR]

The present study was framed with the aim to develop a simple, efficient, and robust reverse-phase HPLC method for the quantification of plumbagin utilizing an analytical quality-by-design (AQbD) approach. HPLC was performed in Waters made high-performance liquid chromatography equipped with Waters 2996 photodiode array detector (detection at 254 nm), and reversed-phase Waters inertsil C18 column. The AQbD approach helps in finding the most suitable critical analytical attributes (CAAs) as well as high-risk critical method parameters (CMPs). Using the Box-Behnken Design experiment, the selected CMPs were optimized for the higher recovery of plumbagin. Further, the developed method was validated following International Conference on Harmonization (ICH) Q2 (R1) guidelines. The low divergence in intra- and inter-day experiments, and low LOD (2.4 µg/mL) and LOQ (7.2 µg/mL) values justify the high precision and sensitivity of the developed method. The developed method was further tested for the quantification of plumbagin from the two different plant source collected from two different climatic conditions. Results show that the maximum plumbagin content was found in P. indica roots. In conclusion, the AQbD approach could be used in other plant systems for the optimization of different variables to obtain a higher amount of targeted plant metabolite. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica is an effective medicinal plant and a potent herb for various diseases. The leaves of P. zeylanica were subjected to crude extraction using Ethyl acetate. Chromatographic analysis of the crude extracts of P. zeylanica were studied in thin layer chromatography on a silica gel matrix. Further the extracts were purified in column chromatography and the fractions were eluted in Ethyl acetate, Chloroform solvent system. The molecules eluted in the fractions were checked in UV-spectroscopy and further characterised by GC-MS analysis. Phenol, 2,4, bis (1,1-dimethyl ethyl)-(7311) highlighted from the chromatographic studies was studied for in silico anticancer effect on matrix metalloproteins, whose dysfunction cause oral squamous cell carcinoma. [ABSTRACT FROM AUTHOR]

Two cDNAs encoding type Ш polyketide synthase (PKS1) and chalcone synthase (CHS, PKS2), were cloned from fresh leaves of Plumbago zeylanica L. (P. zeylanica). Their heterologous expression revealed that PKS1 catalyzed the formation of five α-pyrones from three to six acetate units by accepting acetyl-CoA and malonyl-CoA. In contrast, PKS2 catalyzed the formation of naringenin and bisnoryangonin by accepting p-coumaroyl-CoA and malonyl-CoA. Naringenin is thought to be involved in the biosynthesis of various bioactive flavonoids. PKS2 can be used to molecular breeding to enhance the production of these useful secondary metabolites via its overexpression. [ABSTRACT FROM AUTHOR]

In the present work, a simple, rapid, reliable and accessible high-performance thin-layer chromatography (HPTLC) technique has been established for the concurrent quantification of plumbagin. The well-known medicinal plant, Plumbago zeylanica L., has been traditionally used to treat a variety of ailments in India and other countries. Chromatographic separation was conceded on pre-coated silica gel plates F254. The apposite mobile phase toluene‒acetic acid (9.0:0.5, V/V) was developed to expand the plates which separated components according to the marker compounds. Densitometric scanning was performed by CAMAG Scanner V and measured at wavelength 272 nm. The marker compounds were practically resolved with RF 0.44 ± 0.02 for plumbagin. Further, the antioxidant potential of the methanolic extract of P. zeylanica was also assessed with 1,1-diphenyl-2-picrylhydrazyl (DPPH) by using an ultraviolet (UV) spectrophotometer. The percentage inhibition of free radical was found to be 76.5%. The developed HPTLC method was validated for accuracy, linearity, precision and specificity. The extract has revealed considerable antioxidant activity. The reported existing phenolic and flavonoids compounds are responsible for the antioxidant activity of the plant extract as reported in the literature. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica L. (Pz) is one of the most important medicinal plant belonging to the family Plumbaginaceae. It is a perennial shrub, growing throughout India and most places of Sri Lanka. It contains various bioactive compounds like alkaloids, flavonoids, naphthoquinones, glycoside, saponins, steroids, tri-terpenoids, coumarins, phenolic compounds etc. Of all the chemical constituents, plumbagin is the principal active compound. Plumbagin (5-hydroxy-2-methyl-1, 4-naphthoquinone-C11H8O3) is primarily present in roots in higher amounts with only about 1% in the whole plant. The literature reveals that the root and root bark have a wider claim in traditional medicines against various diseases as a memory enhancer, anti-inflammatory, anti-microbial, wound healing, anti-malarial, anti-infertility, anticancer, blood coagulation, and anti-oxidant activities. The present study aims to evaluate the anti-inflammatory and antimicrobial properties of this plant. [ABSTRACT FROM AUTHOR]

Key message: The present study provides comparative transcriptome analysis, besides identifying functional secondary metabolite genes of Plumbago zeylanica with pharmacological potential for future functional genomics, and metabolomic engineering of secondary metabolites from this plant towards diversified biomedical applications. Plumbago zeylanica is a widely used medicinal plant of the traditional Indian system of medicine with wide pharmacological potential to treat several disorders. The present study aimed to carry out comparative transcriptome analysis in leaf and root tissue of P. zeylanica using Illumina paired end sequencing to identify tissue-specific functional genes involved in the biosynthesis of secondary metabolites, contributing to its therapeutic efficacy. De novo sequencing assembly resulted in the identification of 62,321 "Unigenes" transcripts with an average size of 1325 bp. Functional annotation using BLAST2GO resulted in the identification of 50,301 annotated transcripts (80.71%) and GO assigned to 18,814 transcripts. KEGG pathway annotation of the "Unigenes" revealed that 2465 transcripts could be assigned to 242 KEGG pathway maps wherein the number of transcripts involved in secondary metabolism was distinct in root and leaf transcriptome. Among the secondary metabolite biosynthesis pathways, the cluster of "Unigenes" encoding enzymes of 'Phenylpropanoid biosynthesis pathway' represents the largest group (84 transcripts) followed by 'Terpenoid Backbone biosynthesis' (48 transcripts). The transcript levels of the candidate unigenes encoding key enzymes of phenylpropanoid (PAL, TAL) and flavanoid biosynthesis (CHS, ANS, FLS) pathways were up-regulated in root, while the expression levels of candidate "Unigenes" transcript for monoterpenoid (DXS, ISPF), diterpenoid biosynthesis (SPS, SDS) and indole alkaloid pathways (STR) were significantly higher in leaf of P. zeylanica. Interestingly, validation of differential gene expression profile by qRT-PCR also confirmed that candidate "Unigenes" enzymes of phenylpropanoid and flavonoid biosynthesis were highly expressed in the root, while the key regulatory enzymes of terpenoid and indole alkaloid compounds were up-regulated in the leaf, suggesting that (differences in) the levels of these functional genes could be attributed to the (differential) pharmacological activity (between root and leaf) in tissues of P. zeylanica. [ABSTRACT FROM AUTHOR]

The metabolite contents of the plants may vary with season and geographic variations, and this induces the biological potential of the plant species. The present study aimed to identify the elite chemotype(s) of Plumbago zeylanica, collected from the natural habitat of Gangetic plains on the basis of plumbagin content. The effect of intra-specific variability in the plumbagin content among the population was estimated by using high-performance thin-layer chromatography (HPTLC) densitometric method. Roots of thirteen naturally occurring populations of P. zeylanica from the entire range were collected, covering the wide topography from foothills up to the highest peak. The method developed for quantification of plumbagin was validated and found to be linear, specific and accurate with precision and accuracy. LOD and LOQ were 1.04 and 3.16 ng/spot. Precision studies (both inter-day and intra-day) were within the standard limit of API, with RSD (%) less than 3%. Quantification of plumbagin within population reveals that it varies from 0.0048 ± 0.0005 μg (NBP-22) to 0.1696 ± 0.003 μg (NBC-25) percent on per dry weight basis. Analysis of variance (ANOVA) suggests that there is a significant difference in plumbagin content among the population. UPGMA cluster analyses based on plumbagin content bifurcate the populations into two major clusters, representing the variation in metabolite content. The first major cluster having NBP-25 and NBP-18 accessions, NBP-25 having maximum plumbagin content was collected from Bankura, West Bengal. The study suggests the presence of chemotaxonomic variation among the population which may be due to the change in environmental/phytogeographical factors. [ABSTRACT FROM AUTHOR]

Global emergence and persistence of the multidrug-resistant microbes have created a new problem for management of diseases associated with infections. The development of antimicrobial resistance is mainly due to the sub-judicious and unprescribed used of antimicrobials both in healthcare and the environment. Biofilms are important due to their role in microbial infections and hence are considered a novel target in discovery of new antibacterial or antibiofilm agents. In this article, zinc oxide nanoparticles (ZnO-NPs) were prepared using extract of Plumbago zeylanica. ZnO-NPs were characterized and then their antibiofilm activity was tested against Gram-positive and Gram-negative bacteria. The ZnO-NPs were polydispersed, and the average size was obtained as 24.62 nm. The presence of many functional groups indicated that phytocompounds of P. zeylanica were responsible for the synthesis, capping, and stabilization of ZnO-NPs. Synthesized NPs inhibited the biofilm formation of E. coli, S. aureus, and P. aeruginosa by 62.80%, 71.57%, and 77.69%, respectively. Likewise, concentration-dependent inhibition of the EPS production was recorded in all test bacteria. Microscopic examination of the biofilms revealed that ZnO-NPs reduced the bacterial colonization on solid support and altered the architecture of the biofilms. ZnO-NPs also remarkably eradicated the preformed biofilms of the test bacteria up to 52.69%, 59.79%, and 67.22% recorded for E. coli, S. aureus, P. aeruginosa, respectively. The findings reveal the ability of green synthesized zinc oxide nanoparticles to inhibit, as well as eradicate, the biofilms of Gram-positive and Gram-negative bacteria.

Forty two accessions of Plumbago zeylanica were collected and evaluated for morphological, yield and quality traits. The results revealed that the accession Pz 27 registered the highest mean value for plant height (178.33cm) and fresh weight of root (518.60g).The same accession registered the maximum number of roots (32.33). Plumbagin content ranged from 0.19 - 0.34 % and was maximum in Pz 15 (0.34%) followed by Pz 17 (0.33%). Correlation between dry root weight and morpho - economic characters revealed that dry root weight exhibited positive and highly significant correlation with fresh root weight, root number and plant height. Based on the dry root weight and plumbagin content the accessions in Part III (Pz 19 and Pz 20) and Part IV (Pz 25, Pz 27, Pz 39, Pz 40 and Pz 41) were identified as best performing accessions

Objectives: The current investigation was carried out to determine the cytotoxic and the antimicrobial activities of methanolic extracts of Plumbago zeylanica. Methods: The stems, leaves, and whole plants were air dried and extracted with methanol by using a Soxhlet extractor for 72 hours at 55 - 60°C. The antimicrobial activities were determined from the zones of inhibition, which were measured by using the agar well diffusion method, and the cytotoxicity assays were performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method. Results: The methanolic extracts of the stem and the leaves of Plumbago zeylanica were tested against six bacterial species and nine fungal species, and both extracts showed antimicrobial activity in a dose-dependent manner. The leaf extract of Plumbago zeylanica showed maximum antimicrobial activity against both Staphylococcus aureus sub sp aureus and Fusarium oxysporum. The stem extract was found to be more antimicrobial against the Pseudomonas aeruginosa and the Penicillium expansum species. MTT assays were used to test the cytotoxicity of the whole plant extract in the HCT-116 and the K-562 cell lines, and that extract was shown to have weak cytotoxicity in both cell lines. Conclusion: In the present study, the methanolic stem extracts of Plumbago zeylanica were found to possess remarkable antibacterial activities against many human and agricultural pathogens. The extracts were also found to possess significant antifungal activities, but the antifungal activities were less than the antibacterial activities. Finally, the extracts were found to have weak cytotoxicities in the HCT-116 and the K-562 cell lines.

Traditionally, the leaves of Plumbago Zeylanica were used for the treatment of several diseases. Herein, we report phytochemical analysis, total flavonoid and phenolic content, antioxidant capacity, and antibacterial activities of the leaf extracts of the plant. The phytochemical analysis test of the extracts showed the presence of biologically important compounds such as phenolics, flavonoids, alkaloids, terpenoids, and glycosides. Moreover, the total flavonoid content of 69.566, 66.966, and 59.133 mgQE/100 g was obtained from methanol, ethyl acetate, and petroleum ether extracts respectively. And the total phenolic content of methanol, ethyl acetate, and petroleum ether extracts were 82.738, 69.784, and 51.630 mgGAE/g dry samples. The extracts have promising antioxidant and antibacterial activity against S.aureus (Staphylococcus aureus), S. pyogens (Streptococcus pyogens), E. coli (Escherichia coli), and K. peneumona (Klebsiella pneumonia). The high antibacterial and antioxidant activity was found for methanol extract of the leaf extract of P. Zeylanica is attributing to the presence of polar phytochemicals such as; flavonoids, phenolic, and glycosidic groups in the crude extract obtained from maceration of polar solvents. [ABSTRACT FROM AUTHOR]

Plumbago scandens L. (Plumbaginaceae) occurs in all regions of Brazil. It has been described as toxic to cattle and goats. Caustic lesions in the upper digestive tract characterize poisoning. P. scandens contains a naphthoquinone named plumbagin, which presents high cytotoxic activity. Plumbago auriculata Lam., a widely used ornamental plant, is considered potentially toxic, but there is limited data about its toxicity. This work aimed to validate analytical methodologies for determining the levels of plumbagin in samples of leaves, stems, and rumen content to be used as an auxiliary chemical marker in the laboratory diagnosis of intoxication. One methodology used thin layer chromatography (TLC), and another used high-performance liquid chromatography (HPLC). The presence of palisade grass (Urochloa brizantha (Hochst. ex A.Rich.) R.D.Webster), Guinea grass (Megathyrsus maximus (Jacq.) B.K.Simon & S.W.L.Jacobs), corn silage, and rumen content did not interfere with plumbagin in the two methodologies. The TLC methodology generates qualitative results but is simple to implement and has a low cost. The HPLC methodology showed a limit of detection (LOD) of 0.01 μg/mL and a limit of quantification (LOQ) of 0.05 μg/mL. Leaf and stem samples of P. scandens evaluated showed high levels of plumbagin (0.261 ± 0.087 % and 0.327 ± 0.055 %, respectively). In contrast, leaves of P. auriculata did not show detectable levels of the toxin, and some stem samples showed low levels (up to 0.000114 %). Thus, these methodologies can be used to confirm or rule out the consumption of P. scandens in rumen content from animals suspected of poisoning. [Display omitted] • TLC and HPLC methods for detecting plumbagin from Plumbago scandens in rumen and plant samples were validated. • The leaves and stem of Plumbago scandens showed high levels of plumbagin. • Plumbago auriculata showed no detectable levels in the leaves and low levels in the steams. [ABSTRACT FROM AUTHOR]

We investigated the anticancer activity of the PzMH fraction (hexane fraction) extracted from the roots of Plumbago zeylanica L., an ethnomedicinally significant plant widely distributed in India. The PzMH fraction was obtained through rigorous extraction and purification processes. In vitro cytotoxicity assays were performed to assess its effects on Ehrlich ascites carcinoma (EAC) cells. Acute toxicity studies were conducted to evaluate its safety profile. An in vivo study was carried out on EAC-bearing Swiss albino mice to assess its anticancer efficacy. Flow cytometric and microscopic analyses were done to examine the induction of cell death by the PzMH fraction. Western blot analysis was used to investigate the molecular mechanisms involved. The PzMH fraction exhibited a significant cytotoxic effect in vitro , resulting in 50% cell death in EAC cells at low concentrations. The calculated GI 50 value for the PzMH fraction was 42.74 µg/ml, demonstrating a comparable efficacy to the standard drug 5-fluoro uracil (GI 50 = 43.38 µg/ml). The safety of therapeutic doses was confirmed through acute toxicity studies, which yielded an LD 50 value of 500 mg/kg body weight. In the in vivo study, the PzMH fraction demonstrated a substantial 79.05% inhibition in the growth of EAC cells at the 300 mg/kg body weight dose of PzMH fraction. Flow cytometric and microscopic analyses revealed distinct apoptotic features in EAC cells treated with the PzMH fraction. Cell cycle analysis showed a significant arrest at the G 0 /G 1 stage following treatment. PzMH treatment elicited a response characterized by escalated levels of cleaved caspases-3 and −9, while concurrently leading to a decreased expression of the Bcl2 protein, as evidenced by Western blot analysis. The current research provides empirical evidence supporting the anticancer activity of the PzMH fraction extracted from P. zeylanica. The observed cytotoxicity, safety, and apoptosis-inducing properties make it a promising candidate for further investigation as a potential cancer therapy. Further exploration of its phytochemical composition, including major compounds such as 4-hydroxybenzaldehyde, trans -cinnamic acid, plumbagin and lawsone, contributes to our understanding of its mechanisms of action. [ABSTRACT FROM AUTHOR]

Plasmid curing is the process of obviating the plasmid encoded functions such as antibiotic resistance, virulence, degradation of aromatic compounds, etc. in bacteria. Several plasmid curing agents have been reported in literature, however, no plasmid curing agent can eliminate all plasmids from different hosts. Hence, there is always a need for novel plasmid curing agents that can be effectively used for reversal of plasmid encoded functions such as virulence, antibiotic resistance, etc. In the present study, an active principle responsible for the plasmid curing activity was purified from roots of Plumbago zeylanica by bioassay guided fractionation and identified as 2-hydroxy-1,4-naphthoquinone (lawsone), on the basis of spectral and analytical data such as NMR, GCMS, FTIR. Plasmid curing activity of lawsone was observed against reference as well as wild plasmids (pBR322, pRK2013, R136, pUPI281, and pUPI282) residing in a range of hosts. Curing of plasmid was confirmed by agarose gel electrophoresis. MICs of antibiotics against A. baumannii A24 (pUPI281) and E. coli (pRK2013) decreased significantly in presence of lawsone suggesting synergy between lawsone and antibiotics. Lawsone also inhibited transfer of plasmid pRK2013 to E. coli either by transformation or conjugation. Viability assays (MTT) revealed that lawsone was not toxic to mammalian cells. Thus, the present investigation has revealed lawsone as an effective plasmid curing agent capable of suppressing development and spread of antibiotic resistance. Further, lawsone has important application in basic research to identify phenotypes encoded by the plasmids in plasmid curing experiments. To the best of our knowledge this is the first report of plasmid curing activity of lawsone isolated from roots of P. zeylanica.

Plasmid curing is the process of obviating the plasmid encoded functions such as antibiotic resistance, virulence, degradation of aromatic compounds, etc. in bacteria. Several plasmid curing agents have been reported in literature, however, no plasmid curing agent can eliminate all plasmids from different hosts. Hence, there is always a need for novel plasmid curing agents that can be effectively used for reversal of plasmid encoded functions such as virulence, antibiotic resistance, etc. In the present study, an active principle responsible for the plasmid curing activity was purified from roots of Plumbago zeylanica by bioassay guided fractionation and identified as 2-hydroxy-1,4-naphthoquinone (lawsone), on the basis of spectral and analytical data such as NMR, GCMS, FTIR. Plasmid curing activity of lawsone was observed against reference as well as wild plasmids (pBR322, pRK2013, R136, pUPI281, and pUPI282) residing in a range of hosts. Curing of plasmid was confirmed by agarose gel electrophoresis. MICs of antibiotics against A. baumannii A24 (pUPI281) and E. coli (pRK2013) decreased significantly in presence of lawsone suggesting synergy between lawsone and antibiotics. Lawsone also inhibited transfer of plasmid pRK2013 to E. coli either by transformation or conjugation. Viability assays (MTT) revealed that lawsone was not toxic to mammalian cells. Thus, the present investigation has revealed lawsone as an effective plasmid curing agent capable of suppressing development and spread of antibiotic resistance. Further, lawsone has important application in basic research to identify phenotypes encoded by the plasmids in plasmid curing experiments. To the best of our knowledge this is the first report of plasmid curing activity of lawsone isolated from roots of P. zeylanica. [ABSTRACT FROM AUTHOR]

Study of the chemical constituents of the roots of Plumbago zeylanica L. collected in Taiwan led to the isolation and identification of a new naphthoquinone dimer, plumzeylanone (1), along with eight known compounds (2-9). Nine naphthoquinones isolated from this plant were assayed for cell growth inhibition activity using NALM-6 (human B cell precursor leukaemia), A549 (human lung adenocarcinoma), Colo205 (human colorectal adenocarcinoma) and KB (human epidermoid carcinoma). Plumzeylanone (1), a novel plumbagin dimer, suppressed cell proliferation in only NALM-6 cells (IC50 3.98 μM). However, maritinone (9) showed strong inhibition of cell growth in all cell lines tested (0.12 < IC50 < 9.06 μM). This compound appeared to affect the cell cycle. [ABSTRACT FROM AUTHOR]

This investigation aimed to optimize the extraction of plumbagin by the microwave-assisted extraction (MAE) method and optimize the extraction parameters using response surface methodology (RSM) and adaptive neuro-fuzzzy inference system (ANFIS). A single-factor experiment was conducted to examine the range of the extraction parameters. Box and Behnken's design (BBD) which is a three-level factorial experiment was selected to obtain the best combination of three extraction parameters, namely extraction time, solvent volume, and particle size. The experimental data obtained was analyzed and fitted in the second-order polynomial equation, the r 2 value of 0.993 for plumbagin yield was obtained. The model was found significant and all three parameters had a significant effect on the plumbagin yield. The optimal parameters were extraction time of 4 min, the solvent volume of 20 mL, and particle size 0.6 mm. At these optimal conditions, the plumbagin yield was 0.992% which was found to be close to the predicted values of RSM and ANFIS. The results obtained showed that MAE can be a highly efficient method for plumbagin extraction by reducing time and solvent consumption by many folds. There is a rising demand for plumbagin in the pharmaceutical industry; this study will allow a time-efficient and cost-effective extraction process. [Display omitted] • First report on plumbagin estimation in different parts of Plumbago zeylanica and P auriculata. • Microwave-assisted extraction of plumbagin. • Optimization of microwave-assisted extraction of plumbagin by RSM and ANFIS. [ABSTRACT FROM AUTHOR]