Cytotoxic Activity of Acetone Extracts of Podophyllum peltatum L. Plant Organs and Cell Cultures in Relation to Normal and Tumor Cells

Sinopodophyllum hexandrum (S. hexandrum) is an endangered traditional Chinese medicine as abundant podophyllotoxin with powerful anticancer activity. In this study, the rootstalks of S. hexandrum from different geographical locations in China [S1 (Gansu) and S2 (Shaanxi)] were used as research materials to clone the key gene pluviatolide O-methyltransferase 3 (ShOMT3) in the podophyllotoxin biosynthetic pathway. Subsequently, bioinformatics analysis of the ShOMT3 gene and its encoded protein was subjected to bioinformatics analysis using various analysis software including ProtParam, DeepTMHMM, SubLoc, Signal-P 5.0, and Swiss-model. The results of the analysis revealed that the CDS region of the ShOMT3 gene is 1119 bp long, encoding 372 amino acids. The theoretical molecular weight of the ShOMT3 protein is 41.32784 kD, and the theoretical isoelectric point (pI) is 5.27. The instability coefficient of the protein is 46.05, the aliphatic index is 93.58, and the grand average of hydropathicity (GRAVY) is 0.037, indicating that it is an unstable hydrophobic protein. The protein does not contain transmembrane domains or signal peptides, indicating that it is a non-secreted protein. Secondary structure prediction results suggests that the protein consists of alpha helices, random coils, extended strands, and beta-turns. Tertiary structure prediction results suggests that the protein functions as a monomer. In the phylogenetic tree, the ShOMT3 protein has the highest homology with Podophyllum peltatum (P. peltatum). The successful cloning and bioinformatics analysis of the ShOMT3 gene provide theoretical basis and excellent genetic resources for the molecular regulatory mechanism analysis of the podophyllotoxin biosynthetic pathway and molecular breeding in S. hexandrum.

In the field of oncology, the use of herbal medicines as a supplemental or alternative treatment option has been generally accepted (Catharanthus roseus, Podophyllum peltatum L., Taxus brevifolia Nutt., Taxus baccata, etc.). As a result, several brand-new cytotoxic chemicals are discovered each year in plants, opening up fresh avenues for the treatment of cancer. The examination of naturally occurring molecular entities that could benefit the pharmaceutical business is a focus of many researchers. The search for clinical efficacy validation follows the discovery of drugs with anticancer activity in preclinical trials. Only 29 of the 240 anticancer medicines licensed in the previous 40 years are entirely synthetic, which could be attributed to the advantages of natural substances such as lower side effects and the ability to affect numerous Signaling pathways involved in the carcinogenesis process. In addition, synthetic compounds with natural pharmacophores that imitate the effects of natural products have been licensed as anticancer medications throughout the past ten years. Since the start of cancer research, phytochemicals have been a focal point because they were some of the first antineoplastic medicines found (e.g., leucovorin in 1950, carzinophilin in 1954, vincristine in 1963, actinomycin D in 1964, etc.). Additionally, their research is still ongoing today. It is crucial to note that natural substances are used both as adjuvants and chemotherapeutic agents in the treatment of cancer. By summarizing the three aspects of natural chemicals' anticancer action, this review offers a fresh viewpoint on their use in the field of oncology. Chemotherapeutic drugs because of their inherent antitumor effects, chemopreventive drugs, and sensitizers for multi-drug resistance are the first three categories.

Podophyllotoxin (PPT), a naturally occurring aryltetralin-type lignan obtained from Podophyllin, an ethanolic extract of Podophyllum peltatum L. (syn. P. hexandnum Royle), exhibits marked biological activity as strong antineoplastic drugs and antiviral agents. Chemical transformations performed on PPT resulted in analogs which also display potent cytotoxic, antiviral, and immunosuppressive activities. In the present research, ten new compounds of PPTs 2a-l bearing pyrimidine moiety were synthesized by the reaction of PPT (1) with 2-methyl sulphonyl pyrimidines in the presence of tetrahydrofuran solvent and base sodium hydride. All the synthesized compounds were tested for antimicrobial activities.. KEYWORDS :4,6-Dimethoxy-2-methylsulfonylpyrimidine, Antimicrobial activity, Podophyllotoxin.

Little is known about how carbon integration and storage dynamics affect and are affected by demography in field populations. We sought to elucidate this link by examining dynamic patterns of carbon integration relative to the timing of demographically significant developmental decisions regarding shoot type determination in mayapple, Podophyllum peltatum, a clonal plant with large and persistent rhizomes. Individual rhizome systems growing in natural populations were fed 14CO2 either in late-April, early-May, or mid-June, then harvested at intervals throughout the current season and into the next. When distribution of label was examined we found that carbon fixed at different times in the growing season is used differently: April-fixed assimilate remained in the labeled shoot or was moved into the old rhizome, May-fixed assimilate was found predominantly in the old rhizome, while early-June fixed assimilate moved into the old rhizome and the extending new ramet. Movement of assimilate into the old rhizome appeared to have precedence over formation of additional new ramets. Despite significant within season changes in location of dominant sinks within rhizome systems, there was little redistribution of labeled assimilate: early fixed assimilate was not used to fuel later within season growth, however, assimilate was redistributed between seasons. Vegetative and sexual systems differed in the distribution only of April-fixed assimilate. This was observed even though early labeling occurred prior to anthesis. Sexual systems retained a greater proportion of assimilate in the stem than did vegetative ones, which exported more to the old rhizome. 14C-distribution patterns did not vary between systems differing in future demographic status suggesting that the developmental decision regarding shoot type is based on resources acquired in prior years. We explore the hypothesis that preformation and storage are functionally linked traits that permit plants to coordinate the developmental determination of structures differing in cost and demographic function with known resource status. We conclude that demography influences and is influenced by integrative physiology and that physiological restrictions on within season redistribution of assimilates constrain plants' capacities to respond to short-term environmental variation. Such constraints may affect plants' abilities to respond to rapid environmental change in the Anthropocene.

Podophyllotoxin (PPT) is used in the industrial production of efficient anticancer, antiviral and other drugs. Sinopodophyllum hexandrum or Podophyllum peltatum are natural sources of PPT, but at present they are considered as endangered species. Their PPT content is variable, depending on the growing conditions. Searching for new sources of PPT, some representatives of the genus Juniperus were found to exhibit efficient PPT biosynthesis. However, PPT is highly toxic and poorly soluble in water compound, which limits its clinical applications. In this connection, amphiphilic polymer micelles are considered to be suitable PPT carriers, aimed at increase in water solubility and decrease in toxicity. The present research deals with the evaluation of MPEG-polycarbonate block copolymer micelles loaded with PPT or juniper extracts. The active component-loaded polymer nanocarriers were characterized by dynamic and electrophoretic light scattering, as well as by transmission electron microscopy. The active component loading efficiency and loading capacity were also determined. Highly efficient antiproliferative activity of the loaded micelles was determined in a panel of cancer cell lines. The obtained amphiphilic nanocarriers, loaded with PPT-containing bioactive components, have application in future in vivo preclinical trials of their pharmacokinetics and pharmacodynamics as potential therapeutical agents in the prospective nanomedicine.

Glacial/interglacial cycles and topographic complexity are both considered to have shaped today's diverse phylogeographic patterns of taxa from unglaciated eastern North America (ENA). However, few studies have focused on the phylogeography and population dynamics of wide-ranging ENA herbaceous species occurring in forest understory habitat. We examined the phylogeographic pattern and evolutionary history of Podophyllum peltatum L., a widely distributed herb inhabiting deciduous forests of ENA. Using chloroplast DNA (cpDNA) sequences and nuclear microsatellite loci, we investigated the population structure and genetic diversity of the species. Molecular dating, demographic history analyses, and ecological niche modeling were also performed to illustrate the phylogeographic patterns. Our cpDNA results identified three main groups that are largely congruent with boundaries along the Appalachian Mountains and the Mississippi River, two major geographic barriers in ENA. Populations located to the east of the Appalachians and along the central Appalachians exhibited relatively higher levels of genetic diversity. Extant lineages may have diverged during the late Miocene, and range expansions of different groups may have happened during the Pleistocene glacial/interglacial cycles. Our findings indicate that geographic barriers may have started to facilitate the population divergence in P. peltatum before the Pleistocene. Persistence in multiple refugia, including areas around the central Appalachians during the Quaternary glacial period, and subsequent expansions under hospitable climatic condition, especially westward expansion, are likely responsible for the species' contemporary genetic structure and phylogeographic pattern.

Since times immemorial, mankind has been dependent on Mother Nature in terms of natural resources for alleviating its sufferings. As per W.H.O., developing countries should fully explore their flora and fauna to become economically self-reliant. Country India enjoys having diverse climatic conditions of heavy rainfall, sandy deserts & snow-clad mountains, which play a perfect condition and host for the growth of plenty of medicinal plants with desirable phytoconstituents. Presently mankind is suffering from many diseases that were not so common in the olden days. Does it is a signal from our mother Nature to once again rediscover its treasure of natural resources for exploring answers for various prophylaxis and therapeutic needs of modern-day disorders and diseases? Cancer is the most fatal disease worldwide caused due to the uncontrolled growth of abnormal cells in the body. Ovarian Cancer is the fifth leading cause of death in females. The incidence and mortality rate of Ovarian Cancer is increasing day by day across the world. Biomarkers such as CA125, HE4, OVA1, and Cancer stem cells have been used for the detection of Ovarian Cancer. The current treatment of Ovarian Cancer is surgery and some common treatments include chemotherapy, radiation therapy, adoptive cell therapy, use of medicinal plants and their derivatives. Some prominent medicinal plants like Quercus tinctoria, Curcuma longa, Taxus brevifolia, Allium sativum, Asparagus racemosa, Camptotheca acuminate, Symplocus racemosa, Ginkgo biloba, Genistein, Zingiber officinale, Azadirachta indica, Embilica officinalis, Podophyllum peltatum, Camellia sinensis, Saraca indica. A large number of phytochemicals which has anticancer compounds are flavonoids, alkaloids, terpenoids, carotenoids, phenolics, and organosulfur compounds. Ayurvedic drugs highly effective on Ovarian Cancer are Arka, Bhallataka, Ahipena, Bhanga, Vishamushti, and Jayapala. The diagnosis of Ovarian Cancer in the initial stage is not possible mainly due to a lack of awareness on patients’ behalf. The main effective parameter is imaging for checking the extent and location of the disease. Some other parameters also included physical examination and transvaginal ultrasonography. Thus, the present review is an attempt to compile published reports through authentic sources regarding the current management scenario of Ovarian Cancer with the help of natural drugs or medicinal plants that inhibits or hinders the growth of cancer cell, which may serve as a ready reference for future researchers.

Changes in the Contents of Podophyllotoxin and other Phenolic Compounds in Acetone Extracts of Suspension Cell Cultures of Podophyllum peltatum L. at Different Stages of Cultivation

Podophyllotoxin (PPT) is a precursor for the synthesis of drugs against cancer and other diseases. The present sources of PPT (Sinopodophyllum hexandrum and Podophyllum peltatum) are endangered species, with PPT production highly dependent on their growing conditions. In connection with the identification of new sources of PPT, the present study aimed to recover PPT from Juniperus virginiana leaves via atmospheric or high pressure extraction methods with a focus on using eco-friendly solvents. PPT quantification was determined by UHPLC/HRMS/MS. A thorough study of conventional extraction was carried out to reveal the optimal conditions (solvent ethyl acetate at room temperature and a duration of 1 h) for maximizing the PPT recovery (about 30 mg/g of dry extract and 3 mg/g of dry initial plant material). Peleg’s equation was applied for process kinetics modeling. The best PPT content in the final dry extract (42–45 mg/g of dry extract) was obtained by high pressure methods under supercritical (scCO2 with ethanol or ethyl acetate, 30 MPa, 50 °C and 100 min) or accelerated solvent extraction conditions (solvent ethyl acetate, 10.35 MPa, 20 °C and 3 cycles for 15 min). Seasonal stability and storage stability of the raw material were also determined. The present results have potential applications in the pharmacy for the delivery of PPT from juniper leaves.

Aim: The purpose of this research is to develop a new apple vinegar based on Podophyllum peltatum and analyze the fermentation process using a new methodology. Materials and Methods: The vinegar-making process began with the addition of Saccharomyces cerevisiae (yeast) and l (acetic bacteria) with apple. Chemical titration method is used to find out the apple juice microbial and compared to the microbial in vinegar. The microorganisms Saccharomyces cerevisiae and Acetobacter pasteurianus were found respectively. Fermentation began with fewer Saccharomyces cerevisiae and Acetobacter pasteurianus, but they were present throughout the alcoholic and acetic phases. Both microbial strains produced ethanol and acetic acid using apple as a substrate. The vinegar also produced acetate, volatile alcohols, and aldehydes. Two groups are taken with 9 sample size per group, G power 80%, coincidence intervals will be 95%. Results and Discussion: The innovative apple vinegars had an acetic acid yield of 8.1%. In the sensory evaluation, apple vinegar was well received. The technology proposed here is innovative in that it employs mixed inocula instead of separate ones (p=0.001). It also uses cell biomass instead of centrifugation at the end of the fermentative process. When compared to commercial vinegar (5%), apple vinegar has a high acetic acid concentration (8.1%). Conclusion: Reduce the working time and also be less expensive. Higher acetic acid concentration in vinegar is helpful to improve the taste of the food.

Increases in soil pH and P availability in a temperate hardwood forest affect mycorrhizal colonization and nutrient content of the herbaceous wildflower Podophyllum peltatum (Mayapple)1

Cytotoxic Activity of Extracts from the Intact Plant and Cell Cultures of Podophyllum peltatum Against Cervical Cancer Cells

Phylogenomics and diversification drivers of the Eastern Asian – Eastern North American disjunct Podophylloideae

Abstract In clonal plants, persistent rhizomes can serve multiple purposes, including resource storage, modulation of heterogeneous resource distributions, maintenance of bud banks and promotion of recovery from disturbance. Clonal plants are commonly long‐lived and, in temperate zones, often exhibit organ preformation. Thus, investigations of how the timing of disturbance to the rhizome affects plant performance must occur over multiple growing seasons, but these types of studies are rare. We conducted a field experiment to examine how the persistent rhizome supports the existing shoot, new ramet production and recovery from damage using mayapple Podophyllum peltatum (Berberidaceae), a common herbaceous perennial of low‐light forest understories in Eastern North America. Mayapple maintains a long‐lived rhizome and exhibits a developmentally programmed seasonal pattern of resource transport and new ramet initiation. We varied both the position and timing of rhizome severing in rhizome systems with terminal sexual or vegetative shoots, and tracked plants for 2 years following severing. The location and timing of severing affected both plant persistence (production of new shoots) and performance (leaf area), with effects differing for new shoots that developed at the front versus the back of the rhizome system. Across years, severing location and past years’ shoot size influenced plant persistence and performance, while the effect of timing of severing diminished. Initial sexual status had little effect on rhizome system response that was not accounted for by initial leaf area. Severing generally led to the establishment of two independent rhizome systems. Relative to unmanipulated control systems, these two systems had more total leaf area, but less average leaf area per system. Synthesis. Our results point to the rhizome as a resource integrator that affects plant responses to disturbance immediately following damage and in subsequent growing seasons. Rhizome bud age and/or subtending rhizome size, and developmental programme influence responses to disturbance. While the effects of experimental disturbance on plant performance decreased 2 years after disturbance, further long‐term investigation is needed to fully understand the demographic consequences of damage to persistent rhizomes. A free Plain Language Summary can be found within the Supporting Information of this article.

Podophyllotoxin (PT), a lignan compound from the roots and rhizomes of Podophyllum peltatum, has diverse pharmacological activities including anticancer effect in several types of cancer. The molecular mechanism of the anticancer effects of PT on colorectal cancer cells has not been reported yet. In this study, we sought to evaluate the anticancer effect of PT on human colorectal cancer HCT116 cells and identify the detailed molecular mechanism. PT inhibited the growth of cells and colony formation in a concentration-dependent manner and induced apoptosis as determined by the annexin V/7-aminoactinomycin D double staining assay. PT-induced apoptosis was accompanied by cell cycle arrest in the G2/M phase and an increase in the generation of reactive oxygen species (ROS). The effects of PT on the induction of ROS and apoptosis were prevented by pretreatment with N-acetyl-L-cysteine (NAC), indicating that an increase in ROS generation mediates the apoptosis of HCT116 cells induced by PT. Furthermore, Western blot analysis showed that PT upregulated the level of phospho (p)-p38 mitogen-activated protein kinase (MAPK). The treatment of SB203580, a p38 inhibitor, strongly prevented the apoptosis induced by PT, suggesting that PT-induced apoptosis involved the p38 MAPK signaling pathway. In addition, PT induced the loss of mitochondrial membrane potential and multi-caspase activation. The results suggested that PT induced cell cycle arrest in the G2/M phase and apoptosis through the p38 MAPK signaling pathway by upregulating ROS in HCT116 cells.

Juniper representatives are natural sources of plenty of bioactive metabolites and have been used since ancient times as folk remedies against tapeworms, warts, cancer, etc. The antiproliferative activities of junipers are attributed to podophyllotoxin (PPT), which is a precursor for the synthesis of efficient anticancer drugs. However, the natural sources of PPT, Sinopodophyllum hexandrum (Royle) T. S. Ying and Podophyllum peltatum L., are already endangered species because of their intensive industrial exploitation. Therefore, identification of other sources of PPT is necessary. This study is a broad comparative investigation of junipers, for which original sources have been accessed from different continents of the world. The present research is aimed at the identification of species, producing PPT and other lignans at concentrations that are sufficient for the high antiproliferative activity of the corresponding extracts. Cytotoxic juniper leaf extracts demonstrated a broad spectrum of activity on a panel of cancer cell lines. The antiproliferative properties of junipers were attributed to the combined activity of great diversity of lignans (podophyllotoxin, deoxypodophyllotoxin, β-peltatin, yatein, matairesinol, anhydropodorhizol, etc.), detected by UHPLC-HRMS and LC-ESI-MS/MS in the corresponding extracts. Several species of the genus Juniperus L. were outlined as perspective sources of drug precursors with potential pharmaceutical applications.

Pinoresinol-lariciresinol reductase (PLR) is a key enzyme in podophyllotoxin (PTOX) biosynthesis pathway, which catalyzes the conversion of pinoresinol into secoisolariciresinol, a central precursor for the PTOX biosynthesis. Podophyllum peltatum is an important anticancer species, and PTOX content is obviously different from various plant organs. Therefore, investigation of PLR gene expression facilitates the understanding of the molecular mechanisms of the variation of the PTOX content in different organs from P. peltatum as well as its biosynthesis mechanisms, suggesting that it is an important gene for metabolic engineering of the PTOX. The PTOX was found in all the test organs (root, rhizome, petiole, fruit, leaf and flower), with significant differences among them (p<0.05). The highest PTOX content (2.8018 ± 0.23%) was found in rhizome, which was 30 times higher than the lowest PTOX content in fruit. Furthermore, tissue expression profile showed that P. peltatum PLR (PpPLR) was expressed in all the test organs except for the flower. The highest and lowest expression level was found in rhizome and leaf, respectively. The expression profile of PpPLR was inconsistent with the PTOX content variation in the test organs except for the rhizome, suggesting that the PTOX biosynthetic organ is not necessarily its storage organ. After the initial synthesis, PTOX can be transferred to its storage organ, such as the rhizome.

Cytotoxic activity of extracts obtained from intact plant and callus cell culture Podophyllum peltatum in human leukemic cells

We test whether the invasive earthworm Lumbricus terrestris and leaf litter of the invasive herbaceous plant Alliaria petiolata interact to influence the native plant, Podophyllum peltatum, using both observational field data and a multi-year experiment. We hypothesized invader interactive effects on the native plant might result from either changes in allelochemical distribution in the soil or nutrient availability mediated by the invasive earthworm pulling leaf litter down into the soil. Within the field data we found that Alliaria petiolata presence and higher soil nitrogen correlated with reduced Podophyllum peltatum cover, and no evidence for an invader–invader interaction. Within the factorial experiment, we found a super-additive effect of the two invaders on plant biomass only when activated carbon was present. In the absence of activated carbon, there were no differences in Podophyllum peltatum biomass across treatments. In the presence of activated carbon, Podophyllum peltatum biomass was significantly reduced by the presence of both Lumbricus terrestris and Alliaria petiolata leaf litter. The absence of an effect of Alliaria petiolata leaves without activated carbon, combined with a failure to detect arbuscular mycorrhizal colonization, suggests that indirect effects of allelochemicals on arbuscular mycorrhizal fungi were not the primary driver of treatment responses. Rather direct nutrient availability might influence a potential interaction between these invaders. Leaf nitrogen content was higher and leaf CO2 concentration was lower in the presence of Lumbricus terrestris, but treatment did not influence maximum photosynthetic rate. While the field data do not suggest a negative interaction between these invaders, the experiment suggests that such an interaction is possible with greater environmental stress, such as increasing nitrogen deposition. Further, even plants with rapid physiological responses to increased nitrogen availability may have other physiological limits on growth that prevent them from compensating from the harm caused by multiple invaders.