Paris Polyphylla Research Articles

Qualitative and quantitative analysis of Paris polyphylla var. chinensis by UPLC-Q-TOF-MS/MS and HPLC

Paridis Rhizoma(PR) is prepared from the dried rhizome of Paris polyphylla var. yunnanensis(PPY) or P. polyphylla var. chinensis(PPC) in Liliaceae family. The rapid development of PPY or PPC planting industry resulted from resource shortage has caused the waste of a large number of non-medicinal resources. To clarify the chemical compositions in rhizomes, fibrous roots, stems, leaves, seeds and pericarps of PPC, and explore the comprehensive application value and development prospect of these parts, the qualitative and quantitative analyses on the different parts of PPC were carried out by ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) and high performance liquid chromatography(HPLC). A total of 136 compounds were identified, including 112 steroidal saponins, 6 flavonoids, 11 nitrogen-containing compounds and 7 phytosterols. Rhizomes, fibrous roots, and seeds mainly contained protopennogenyl glycosides and pennogenyl glycosides; leaves and stems mainly contained protodiosgenyl glycosides and diosgenyl glycosides; pericarps mainly contained pennogenyl glycosides, followed by diosgenyl glycosides. The total level of four saponins was the highest in fibrous roots and rhizomes, followed by those in the pericarps and arillate seeds, and the lowest in the stems and exarillate seeds. This study can provide data support for the comprehensive development and rational application of non-medicinal parts of PPC.

Study on the identification and evaluation of growth years for Paris polyphylla var. yunnanensis using deep learning combined with 2DCOS

Paris polyphylla var. yunnanensis, as perennial plants, its quality is closely related to growth period. Different harvest years determine the dry matter accumulation of its medicinal parts and the dynamic accumulation of active ingredients, as well as its economic value and medicinal value. Therefore, it is necessary to establish a systematic evaluation method for the identification and evaluation of P. polyphylla var. yunnanensis with different growth years. Deep learning has a powerful ability in recognition. This study extends it to the identification analysis of medicinal plants from the perspective of spectrum. For the first time, two-dimensional correlation spectroscopy (2DCOS) based on the attenuated total reflection Fourier transformed infrared spectroscopy (ATR-FTIR) combined with residual neural network (Resnet) was used to identify growth years. 525 samples were collected, 4725 2DCOS images were drawn, and the dry matter accumulation in rhizomes of different growth years and different sampling sites were briefly analyzed. The results show that the eight-year-old P. polyphylla var. yunnanensis in Dali has higher economic value and medicinal value. The synchronous 2DCOS models based on ATR-FTIR can realize the identification of growth years with accuracy of 100%. Synchronous 2DCOS are more suitable for the identification of medicinal plants with complex systems. 2DCOS images with different colors and second derivative processing cannot optimize the modeling results. In summary, the method we established is innovative and feasible. It not only solved the identification of growth years, expanded the application field of deep learning, but could also be extended to further research on other medicinal plants.

Polyphyllin Ⅲ-Induced Ferroptosis in MDA-MB-231 Triple-Negative Breast Cancer Cells can Be Protected Against by KLF4-Mediated Upregulation of xCT.

Ferroptosis, which is characterized by the accumulation of intracellular iron and subsequent lipid peroxidation, is a newly discovered form of regulated cell death and plays an important role in tumor suppression. Herein, we showed that Polyphyllin III, which is a major saponin extracted from Paris polyphylla rhizomes, exerted its proliferation-inhibitory effect on MDA-MB-231 triple-negative breast cancer cells mainly through ACSL4-mediated lipid peroxidation elevation and ferroptosis induction. ACSL4 deletion partly attenuated Polyphyllin III-induced ferroptosis. Polyphyllin III treatment also induced KLF4-mediated protective upregulation of xCT, which is the negative regulator of ferroptosis. Interestingly, combination with the xCT inhibitor sulfasalazine (SAS) or downregulation of KLF4 sensitized MDA-MB-231 cells to Polyphyllin III. Furthermore, in vivo xenograft models, SAS significantly sensitized MDA-MB-231 breast cancer cells to Polyphyllin III, likely by enhancing intracellular lipid peroxidation and ferroptosis. The results of this study collectively demonstrated that Polyphyllin III exerts its anticancer effect by inducing ferroptosis via ACSL4 in MDA-MB-231 breast cancer cells. More importantly, we observed for the first time that KLF4-mediated xCT upregulation serves as negative feedback during ferroptosis progression, which might contribute to drug resistance in cancer treatment.

Complete genome sequence analysis of a novel coguvirus isolated from Paris polyphylla var. yunnanensis.

A novel negative-stranded (ns) RNA virus tentatively named "Yunnan paris negative-stranded virus" (YPNSV), was isolated from Paris polyphylla var. yunnanensis plants exhibiting leaf chlorosis and mosaic symptoms in Yunnan. Its complete genome sequence was determined using Illumina and Sanger sequencing. YPNSV has a bipartite genome that consists of a negative-stranded (ns) RNA1 encoding the viral RNA-dependent RNA polymerase (RdRp, p251), an ambisense RNA2 coding for the putative movement protein (MP, p46) and nucleocapsid protein (NP, p39), with the two open reading frames separated by a long intergenic region that is rich in A and U. Sequence comparisons showed that the RdRp, MP, and NP of YPNSV are most similar to those of watermelon crinkle leaf-associated virus 2 (WCLaV-2), with 69.1%, 50.4%, and 60.9% amino acid sequence identity, respectively. Phylogenetic analysis based on deduced amino acid sequences of RdRp and NP showed that YPNSV clustered in a clade with coguviruses and that its closest known relative is WCLaV-2. Based on the above results, YPNSV should be regarded as a new member of genus Coguvirus, family Phenuiviridae.

Gibberellin A3 induces polyaerial shoot formation and increases the propagation rate in Paris polyphylla rhizomes

Paris polyphylla var. yunnanensis is a rhizomatous geophyte in high demand for industrial and medicinal purposes. It grows very slowly, and thus, rhizome cutting is a promising propagation technique. However, whether and how polyapical shoots can be artificially produced with rhizomes is unclear. This study reported that a P. polyphylla rhizome mainly sprouts one shoot but could occasionally grow polyapical shoots. Application of phytohormones, including auxins, cytokinins, abscisic acids and gibberellins on P. polyphylla rhizomes showed that GA3 could break dormancy in two weeks and induced polyapical shoots at a rate of 100 %. The apical shoot number per GA3-treated rhizome reached 7.2. The GA3-induced occurrence of polyapical shoots derived from the repetitive de novo formation of apical buds. GA3 application changed endogenous auxin, cytokinin or ABA concentrations slightly or not at all but dramatically increased endogenous GA3 and GA1 levels in rhizomes. Transcriptional profiling showed there were few shared differentially expressed genes and significant differences in the expression patterns of hormone-related genes between rhizomes with mono- and polyapical shoots. These results suggest that P. polyphylla has the genetic potential to produce polyapical shoots. GA3 treatment can quickly break dormancy and strongly induce polyapical shoots in this species, providing an efficient rhizome cutting technique for P. polyphylla propagation.

Polyphyllin I reverses the resistance of osimertinib in non-small cell lung cancer cell through regulation of PI3K/Akt signaling

Lung cancer is considered the main cause of cancer mortality worldwide. Osimertinib, a third-generation EGFR-TKI, has been approved and administrated for treating patients with either EGFR T790M mutation or EGFR sensitive mutation. However, resistance to osimertinib emerges and has been considered to be the main obstacle in lung cancer treatment. Polyphyllin I is isolated from the natural herb Paris polyphylla and exhibits anti-cancer activities. In the present study, we identify Polyphyllin I to reverse the resistance of osimertinib in vitro and in vivo. The results showed that Polyphyllin I reversed the resistance of osimertinib through promoting apoptosis, modulating the PI3K/Akt signaling, and regulating the expression of apoptosis-related proteins in osimertinib-resistant cell lines. In vivo study confirmed the results, showing that the tumor growth was significantly suppressed in the Polyphyllin I/osimertinib group compared to the osimertinib group. It has been clarified that Polyphyllin I could reverse the resistance of osimertinib in osimertinib-resistant non-small cell of lung cancer in vitro and in vivo. The underlying mechanism might be related to the downregulation of the PI3K/Akt signaling and increase of the expression of apoptosis-related proteins, suggesting that Polyphyllin I was a promising therapeutic agent for reversing the resistance of osimertinib.

DNA barcoding aids in identification of adulterants of Trillium govanianum Wall. ex D. Don

The correct scientific identification of the plant material is crucial to the safety and efficacy of herbal products. Trillium govanianum Wall. ex D. Don, a pharmaceutically prized medicinal plant species endemic to the Himalaya, has been recently subjected to large-scale extraction in the wild due to increasing market demand. Consequently, adulteration and/or substitution of the traded material of T. govanianum with closely-related plants such as Paris polyphylla Sm. has become a quality control problem for the herbal industry. Therefore the present study aimed to (i) develop a reference DNA barcode of T. govanianum and its potent adulterant P. polyphylla for correct identification, (ii) and check adulteration of market samples of T. govanianum. Reference DNA barcode library of T. govanianum and P. polyphylla was successfully established, using Internal transcribed spacer (ITS), Maturase K (matK), chloroplast intergenic spacer (trnH-psbA), and ribulose-bisphosphate carboxylase (rbcL) regions. The ITS, matK and trnH-psbA were found to be ideal reference barcodes for T. govanianum, while ITS and trnH-psbA were suitable for P. polyphylla. All the trade samples (i.e. dried rhizomes), investigated during the present study, got putatively identified with the respective reference barcodes; and thereby indicating the ITS and trnH-psbA regions as the potential DNA barcodes for the identification of the trade samples/adulterants. The ITS region showed the highest mean intra- and inter-specific distances, which proves its high efficiency in differentiating closely related species. Phylogenetic trees were also constructed following the neighbor-joining (NJ) method, based on ITS, rbcL, and trnH-PsbA regions of Trillium and Paris species, which distinguished Trillium species from those of Paris. The availability of a novel DNA barcode for this important medicinal plant species will be helpful for correct identification of its raw plant material, checking illegal trade, and to regulate its sustainable collection from the natural habitats.

Tissue distribution, metabolism and absorption of Rhizoma Paridis Saponins in the rats

Ethnopharmacological relevanceParis polyphylla var yunnanensis as a traditional Chinese medicine has been used in the treatment of liver disease for thousands of years. Rhizoma Paridis saponins (RPS) were the main active ingredients in Paris polyphylla with an excellent antitumor effect. However, metabolic and distribution of RPS has not been known.Aim of the study: The objective of this study was to research metabolic and distribution of RPS. Materials and methodsIn this study, the separation and simultaneous determination of RPS in rat plasma and tissues were developed and validated by LC–MS/MS. The permeability and recovery of RPS were tested by Caco-2. S9 assay suggested the metabolic mode of RPS in rats. ResultsAfter oral administration of RPS, the metabolic compound like diosgenin was detected in different tissues although there was none in RPS. The concentration of PI, PII, PVI, PVII, PH and gracillin in the spleen was the highest among these organs. The content of diosgenin were the highest in lung and brain. Caco-2 test indicated that PI, PII, PVI and PVII were low permeability and low recovery. Efflux ratio indicated that PVI should be a potential P-gp substrate. Potential P-gp substrate may be PVI. S9 assay suggested that RPS possess slow metabolic and moderate metabolic compounds. ConclusionsIntegrated LC–MS/MS analysis of serum samples, together with Caco-2 and S9 assays provided a theoretical basis for the application of RPS in the future.

Molecular-networking-guided discovery of species-specific markers for discriminating five medicinal Paris herbs

BackgroundParidis Rhizoma (PR) is a famous traditional herbal medicine. Apart from two officially recorded species, viz. Paris polyphylla Smith var. yunnanensis (Franch.) Hand. - Mazz. (PPY) and P. polyphylla Smith var. chinensis (Franch.) Hara (PPC), there are still many other species used as folk medicine. It is necessary to understand the metabolic differences among Paris species. PurposeTo establish a strategy that can discover species-specific steroidal saponin markers to distinguish closely-related Paris herbs for quality and safety control. MethodsA new strategy of molecular-networking-guided discovery of species-specific markers was proposed. Firstly, the ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) was applied to obtain the MS and MS/MS data of all samples. Then, molecular networking (MN) was created using MS/MS data to prescreen the steroidal saponins for subsequent analysis. Next, the principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA) models were established to discover potential markers. Finally, the verification, identification and distribution of chemical markers were performed. ResultsA total of 126 steroidal saponins were screened out from five species using MN. Five species were classified successfully by OPLS-DA model, and 18 species-specific markers were discovered combining the variable importance in the projection (VIP) value, P value (one-way ANOVA) and their relative abundance. These markers could predict the species of Paris herbs correctly. ConclusionThese results revealed that this new strategy could be an efficient way for chemical discrimination of medicinal herbs with close genetic relationship.

Screening and identification of potassium-dissolving bacteria from different rhizosphere soil of Paris polyphylla var. yunnanensis

The study aiming at exploring the potassium-dissolving capacity of rhizosphere potassium-dissolving bacteria from diffe-rent sources and screen the strains with high potassium-dissolving ability, so as to lay a theoretical foundation for cultivation and quality improvement of Paris polyphylla var. yunnanensis sources. The rhizosphere soil of 10 wild and transplanted species from Yunnan, Sichuan and Guizhou provinces was used as the research object. Potassium-dissolving bacteria were isolated and purified, and their potassium-dissolving capacity was determined by flame spectrophotometry, and identified by physiological, biochemical and molecular biological methods. Twenty-six potassium-dissolving bacteria were purified and 13 were obtained from wild and transplanted strains respectively. It was found through the determination of potassium-dissolving capacity that the potassium-dissolving capacity of 26 strains was significantly different, and the mass concentration of K~+ in the fermentation broth were 1.04-2.75 mg·L~(-1), the mcentration of potassium were 0.01-1.82 mg·L~(-1). The strains were identified as Bacillus, Agrobacterium rhizome and Staphylococcus by physiological, biochemical and 16 S rDNA molecular methods, among them Bacillus amylolyticus(4 strains) was the dominant bacterium of Bacillus. The physiology and biochemistry of rhizosphere potassium-dissolving bacteria in P. polyphylla var. yunnanensis rhizosphere were diffe-rent, and the living environment were different, so the potassium-dissolving capacity also changed. Strain Y4-1 with the highest potassium decomposability was Bacillus amylolytic with a potassium increase of 1.82 mg·L~(-1). The potassium-dissolving ability and the distribution of potassium-dissolving bacteria were different in various habitats. The screening of potassium-dissolving bacteria provided a new strain for the preparation of microbial fertilizer. It is expected that B. amyloidococcus Y4-1 can be used as an ideal strain to cultivate mycorrhizal seedlings of P. polyphylla var. yunnanensis.

Phylogeography of Paris poliphylla var. yunnanensis based on chloroplast gene trnL-trnF sequences

Phylogeography is a research hotspot in the field of the genetic diversity and core germplasm construction of endangered rare plants. Paris polyphylla var. yunnanensis is a rare plant species mainly distributed in China. Wild individuals have been overexploited for the last few decades because of increasing demand for such medicines. Therefore, it is great significance to study the phylogeography of P. poliphylla var. yunnanensis based on chloroplast gene trnL-trnF sequences. In this study, chloroplast genes trnL-trnF were used in the phylogeography analysis of 15 wild and 17 cultivated populations of P. polyphylla var. yunnanensis. This study revealed that based on the results of neutrality tests and mismatch analysis, the rapid expansion of wild population has not been detected in P. polyphylla var. yunnanensis. After aligning and sorting the obtained cpDNA sequences, a total of 15 haplotypes were detected in all 32 populations. One haplotype was unique to the wild population, and 5 haplotypes were unique to the cultivated population. It can be seen that the haplotype richness of cultivated population was higher than that of wild population. The wild populations of P. polyphylla var. yunnanensis were divided into two groups according to evolutionary relationship of haplotypes and distribution map of haplotypes. The haplotype of branch Ⅰ was mainly distributed in Guizhou, and the haplotype of branch Ⅱ was located in Yunnan and Huidong, Sichuan. Therefore, it's speculated that Guizhou and the west Yunnan region may be glacial refuge in the evolutionary history of wild populations of P. polyphylla var. yunnanensis, and in order to protect the wild resources more effectively, wild populations of P. polyphylla var. yunnanensis in these two areas should be included in the protection zone.

Geographical Variation and Correlation Analysis of the Main Bioactive Components of Paris polyphylla from Different Provenances

Geographical Variation and Correlation Analysis of the Main Bioactive Components of Paris polyphylla from Different Provenances

Pharmacokinetics profiles of polyphyllin II and polyphyllin VII in rats by liquid chromatography with tandem mass spectrometry.

Polyphyllin II (PII) and polyphyllin VII (PVII) are the main active ingredients in Paris Polyphylla with an excellent antitumor effect in vitro and in vivo. In this study, a rapid and precise LC-MS/MS method was developed and validated for the separation and simultaneous determination of PII and PVII in rat plasma, tissues, feces and urine using ginsenoside Rg3 as the internal standard. Positive linearity ranged from 1 to 1,000 ng/ml in samples. At the same time, intra- and inter-day precisions were in range of 1.8-12.0%. The accuracy ranged from 95.9 to 100.8%. Mean extraction recoveries of PII and PVII ranged from 86.6 to 96.4%. The analytical method has been successfully applied to the pharmacokinetic studies of PII and PVII in rats after their i.v. administration. After entering systemic circulation, PII and PVII were rapidly distributed in organs, mainly including liver, lung and spleen. Their elimination rate was slow. All of these data provided a theoretical basis for the application of PII and PVII in the treatment of liver- and lung-related diseases.

Factors that influence the plant use knowledge in the middle mountains of Nepal.

An account of total of 58 plant species including 57 genera and 43 families was reported as useful in ethnomedicine from semi-structured questionnaire survey to the 76 participants of Kaski and Baitadi districts, Nepal. Fieldwork and participatory meetings were carried out between September 2017 and January 2018. A total of 419 emic use reports including 150 from Kaski and 269 from Baitadi were reported from 58 ethnomedicinal plant species. Each species was reported for 2-43 use reports and each participant recorded 1-12 use reports. About 25% (n = 104) use reports were associated with the treatment of digestive system disorders followed by 83 for general complaints. Of the species assessed, 53 species had IASc value < 0.25 and only five species had > 0.25. Species Swertia chirayita, Paris polyphylla, Bergenia ciliata, Valeriana jatamansi and Centella asiatica with > 0.25 IASc were found to be highly consented; however they were incongruent between the sample groups and sites. Divergent plant use knowledge specific to each sample district and group was corresponding to the heterogeneity of socio-economy and culture of the sites. Gender, ethnicity, household economy and food availability of the respondents were leading factors affecting the plant use knowledge. Despite the sites were relatively homogenous in eco-physiography, they possessed the distinct plant use knowledge, hinted that the socio-economic factors are more explanatory in plant use knowledge.

Isolation and identification of phosphatolytic bacteria in Paris polyphylla var. yunnanensis

The wild resources of Paris polyphylla var. yunnanensis, a secondary endangered medicinal plant, are severely scarce. Introduction and cultivation can alleviate market demand. To screen phosphatolytic bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis and provide data support for the development of high-efficiency microbial fertilizer, in this study, the dilution plate coating method was used to isolate and screen the phosphorus solubilizing bacteria with the ability of mineralizing organic phosphorus from the rhizosphere soil of wild and transplanted varieties of P. polyphylla var. yunnanensis in 10 different locations in Yunnan, Sichuan and Guizhou. After separation and purification, the phosphatolytic capacity was analyzed by qualitative and quantitative analysis. Combined with physiological and biochemical experiments, the strains were identified using 16 S rDNA sequencing analysis. Forty one strains were selected from the rhizosphere soil of P. polyphylla var. yunnanensis from 10 different habitats. Among them, 21 strains were obtained from the rhizosphere soil of the wild variety P. polyphylla var. yunnanensis and 20 strains were obtained from the rhizosphere soil of the transplanted variety. And significance analysis found that 41 organophosphate solubilizing strains had significant differences in their ability to solubilize phosphorus. The amount of phosphate solubilizing was 0.08-67.61 mg·L~(-1), the pH value was between 4.27 and 6.82. The phosphatolytic amount of strain Y3-5 was 67.61 mg·L~(-1), and the phosphorus increase amount was 57.57 mg·L~(-1). All 41 strains were identified as Gram-positive Bacillus. Combining physiological characteristic and phylogenetic trees, Bacillus mobilis Y3-5 was finally selected as the candidate rhizosphere phosphatolytic bacteria of P. polyphylla var. yunnanensis. The distribution of phosphorus solubilizing bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis was different, and there were significant diffe-rences in phosphorus solubility. Organophosphate-dissolving strain Y3-5 is expected to be a candidate strain of P. polyphylla var. yunnanensis microbial fertilizer.

Influence of drying process on furostanoside and spirostanoside profiles of Paridis Rhizoma by combination of HPLC, UPLC and UPLC-QTOF-MS/MS analyses

Drying method is one of the important factors affecting quality of traditional Chinese medicine. To study the effect of shaded drying and hot air drying on steroidal saponins of Paridis Rhizoma (PR), high performance liquid chromatography (HPLC) analysis was used to investigate the difference of Paris polyphylla var. chinensis (PPC) samples treated by different methods, and then, a rapid and reliable ultra-high performance liquid chromatography (UPLC) method was established to quantitatively analyze the content change of ten steroidal saponins. Hot air drying at 50 ℃ could obviously improve the content of polyphyllin Ⅶ, 17-hydroxygracillin and polyphyllin H, which were major steroidal saponins in PPC. Based on that, the main component changes induced by different drying methods were further analyzed using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS), and the structural identification of varied components revealed that hot air drying could promote the transformation of proto-pennogenyl glycosides to pennogenyl glycosides. This phenomenon was also found in other plants of genus Paris rich in diosgenyl glycosides. The present study provided a useful method for improving quality of PR and valuable information for TCM containing steroidal saponins.

Phytosociological and niche distribution study of Paris polyphylla smith, an important medicinal herb of Indian Himalayan region

The prediction of species distribution is emerging as an important tool for conservation and rehabilitation planning of economically important plant species including medicinal and aromatic plants. Paris polyphylla, an economically and high valued medicinal plant of Indian Himalayan Region (IHR) is reported to lose its habitat area due to over-exploitation. In this study, attempts were made to understand vegetation pattern and find the potential distribution of Paris polyphylla, using MaxEnt in Pindar Valley (2000–4000 m ASL), Bageshwar district of Uttarakhand state, India. MaxEnt is a machine learning data mining technique that estimates a species distribution by finding the distribution of maximum entropy, based on existing knowledge. Phytosociological studies revealed the varying density of target species ranging from 0.50 ind/m2 (P1; 2000 m asl) to 2.00 ind/m2 (P6; 2500 m asl). Among the different geographical provinces of Uttarakhand, MaxEnt predicted wider potential distribution of P. polyphylla in temperate forest with an overall estimated area of 790.85 km2 as suitable. It was found that Precipitation, Elevation and Vegetation type were the most important variables for predicting the habitat suitability. Dwali (ca. 2500 m ASL) site was predicted as a potential distribution area of P. polyphylla with ground-based validation and further suggested this area as a potential reintroduction site. The results of this study can be used to plan protection and re-introduction of P. polyphylla in the region. MaxEnt approach is thus a very promising tool in species conservation planning and in predicting the potential distribution of other high valued medicinal plants of IHR.

Evaluation of antioxidant, anti-inflammatory and anticancer activities of diosgenin enriched Paris polyphylla rhizome extract of Indian Himalayan landraces

Ethnopharmacological relevanceTraditional medicinal plants have gained attention as a potential therapeutic agent to combat cancer and inflammation. Diosgenin rich fresh extracts of Paris polyphylla rhizome from Indian Himalaya is traditionally used as wound healing, anti-bleeding, anti-inflammatory and anti-cancer agent by the folk healers. Aim of the studyPresent study was aimed to prepare two types of extracts from Paris polyphylla rhizome of Indian Himalayan landraces – 1. ethanolic extract of Paris polyphylla rhizome (EEPPR) and 2. Diosgenin enriched Paris polyphylla rhizome extract (DPPE), quantification of diosgenin content, and to evaluate their in vitro anti-oxidant, in vivo anti-inflammatory and in vitro cytotoxicity and anti-cancer activities of the DPPE. Materials and methodsDiosgenin content of EEPPR was quantified through GC-MS while diosgenin content of DPPE was quantified through HPTLC, and the diosgenin yield from EEPPR and DPPE were compared. In vitro antioxidant activities of DPPE were performed using DPPH, NOD, RP and SOD assay while in vivo anti-inflammatory activity of DPPE were evaluated in dextran induced hind paw edema in rats. In vitro cytotoxicity and anti-cancer activities of DPPE were evaluated in human breast cancer cell lines (MCF-7, MDA-MB-231), cervical cancer cell lines (HeLa) and Hep-2 cell lines. ResultsEEPPR obtained through cold extraction method using 70% ethanol showed maximum diosgenin content of 17.90% quantified through GC-MS while similar compounds pennogenin (3.29%), 7β-Dehydrodiosgenin (1.90%), 7-Ketodiosgenin acetate (1.14%), and 7 β-hydroxydiosgenin (0.55%) were detected in low concentration, and thus confirmed diosgenin as major and lead phytochemical. However, DPPE obtained through both cold and repeated hot extraction with the same solvent (70% ethanol) showed diosgenin content of 60.29% which is significantly higher (p < 0.001) than the diosgenin content in EEPPR. DPPE demonstrated significant in vitro antioxidant activities by dose-dependently quenched (p < 0.001) SOD free radicals by 76.66%, followed by DPPH (71.43%), NOD (67.35%), and RP (63.74%) at a max concentration of 2 μg/μl of ascorbic acid and test drugs with remarkable IC50 values (p < 0.01). Further, DPPE also showed potent anti-inflammatory activities by dose-dependently suppressed dextran induced paw edema in rats (p < 0.01) from 2 h to 4 h. DPPE suppressed the proliferation of MCF-7, MDA-MB-231, Hep-2 and HeLa cell lines. Maximum activity was observed in MCF-7 cells. The DPPE also induced apoptosis in MCF-7 cell lines as measured by AO/PI and DAPI staining, as well as DNA laddering, cell cycle analysis and phosphatidylserine externalization assay. The growth-inhibitory effect of DPPE on MCF-7 breast cancer cells was further confirmed from the colony-formation assay. DPPE upregulated expression of Bax and downregulated Bcl-2 and survivin mRNA transcripts. ConclusionDPPE obtained through both cold and repeated hot extraction using ethanol showed significantly higher content of diosgenin than the diosgenin content detected in EEPPR. However, diosgenin yield of both the extracts (EEPPR & DPPE) clearly confirmed diosgenin as major and lead phytochemical of Paris polyphylla rhizome of Indian Himalayan landraces. Further, DPPE also demonstrated potent in vitro anti-oxidative and in vivo anti-inflammatory activities and showed in vitro cytotoxicity and significant anti-cancer (apoptosis) effects in MCF-7 breast cancer cells.

Evaluation of Trace Element Contamination and Health Risks of Medicinal Herbs Collected from Unpolluted and Polluted Areas in Sichuan Province, China

Trace element contamination in Chinese herbal medicines has been recognized as a potential health concern for consumers. To assess the health risk to the herb-consuming population, nine trace elements (Cu, Cd, Cr, Mo, Ni, Pb, Sr, Zn, and As) were investigated based on their concentrations in three common medicinal plants (Astragalus membranaceus, Codonopsis tangshen, and Paris polyphylla var. chinensis) and soils from unpolluted and polluted areas in the Sichuan Province, China. The results showed that the metal content differed significantly in medicinal plants and soils from unpolluted versus polluted areas. No significant differences in metal accumulation were observed for these CHMs grown in either unpolluted or polluted areas. Evaluation of the health risk index suggested that soil ingestion and medicated diet represented the dominant exposure routes, indicating that trace metal(loids) in local soil might pose potential risks through soil-food chain transfer. Hazard quotient values for AM (1.473) and CT (1.357) were higher than the standard value (HQ > 1), whereas the hazard indices for PC, AM, and CT were 13.18, 14.33, and 14.01 times higher than the safe limit (HI > 1) in the polluted area, indicating non-cancer-related health hazards. Ingestion of soil was responsible for 36.39 to 91.06% of the total cancer risk and medicated diet accounted for 6.35 to 62.71%, compared with inhalation and dermal contact, suggesting carcinogenic health risks in herbs from polluted soils. In this study, Pb showed relatively higher non-carcinogenic risks, while Cr and Ni posed the highest cancer risks. Therefore, we propose more effective measures, which should be considered for Cr, Ni, and Pb remediation in soil to reduce their pollution in the studied areas.

Mycorrhizal Fungal Effects on Growth, Antioxidant Capacity, and Medicine Quality of Paris polyphylla var. yunnanensis

A field experiment was conducted to determine the effects of two commercial strains composed of mulple arbuscular mycorrhizal fungi (AMF) species on plant growth, antioxidant capacity, and medicine quality of <i>Paris polyphylla</i> var. <i>yunnanensis</i> in three subtropical soils from Wanzhou, Anshun and Baoshan in fields. The results showed that AMF inoculation enhanced the fungal colonization rate and activities of both succinate dehydrogenase and alkaline phosphatase, thereby, enhancing the mycorrhizal viability of <i>P. polyphylla</i> var. <i>yunnanensis</i>. The concentrations of photosynthetic pigments (chlorophyll <i>a</i>, <i>b</i>, <i>a</i>+<i>b</i> and carotenoids), soluble sugar, soluble protein and photosynthetic capacity were higher in AMF-inoculated plants than in non-AMF-treated plants in field. AMFtreated plants recorded higher activities of catalase, peroxidase and superoxide dismutase, and caused the reduction in malondialdehyde content, indicating lower oxidative damage, compared with non-AMF plants. Polyphyllin I, Polyphyllin II, Polyphyllin III, Polyphyllin IV and total polyphyllin contents were increased by AMF treatment. In conclusion, AMF improved the plant growth, antioxidant capacity and medicinal quality of <i>P. polyphylla</i> var. <i>yunnanensis</i> seedlings. Hereinto, AMF effects on the soil from Wanzhou was relatively greater than on other soils.