Rauwolfia polysaccharide can inhibit the progress of ulcerative colitis through NOS2-mediated JAK2/STAT3 pathway.

BACKGROUND: Ulcerative colitis (UC) is an inflammatory disease of the digestive tract. Rauwolfia polysaccharide (Rau) has therapeutic effects on colitis in mice, but its mechanism of action needs to be further clarified. In the study, we explored the effect of Rau on the UC cell model induced by Lipopolysaccharide (LPS). METHODS: We constructed a UC cell model by stimulating HT-29 cells with LPS. Dextran sodium sulfate (DSS) was used to induce mice to construct an animal model of UC. Subsequently, we performed Rau administration on the UC cell model. Then, the therapeutic effect of Rau on UC cell model and was validated through methods such as Cell Counting Kit-8 (CCK8), Muse, Quantitative real­time polymerase chain reaction (RT-qPCR), Western blotting, and Enzyme-linked immunosorbent assay (ELISA). RESULTS: The results showed that Rau can promote the proliferation and inhibit the apoptosis of the HT-29 cells-induced by LPS. Moreover, we observed that Rau can inhibit the expression of NOS2/JAK2/STAT3 in LPS-induced HT-29 cells. To further explore the role of NOS2 in UC progression, we used siRNA technology to knock down NOS2 and search for its mechanism in UC. The results illustrated that NOS2 knockdown can promote proliferation and inhibit the apoptosis of LPS-induced HT-29 cells by JAK2/STAT3 pathway. In addition, in vitro and in vivo experiments, we observed that the activation of the JAK2/STAT3 pathway can inhibit the effect of Rau on DSS-induced UC model. CONCLUSION: In short, Rauwolfia polysaccharide can inhibit the progress of ulcerative colitis through NOS2-mediated JAK2/STAT3 pathway. This study provides a theoretical clue for the treatment of UC by Rau.

Regulation of hippo signaling mediated apoptosis by Rauvolfia tetraphylla in triple-negative breast cancer.

Rauvolfia tetraphylla is an essential medicinal plant that has been widely used in traditional medicine for various disease treatments. However, the tumor suppressor activity of R. tetraphylla and its phytocompounds were not explored against triple-negative breast cancer. The current research investigated the impact of R. tetraphylla methanolic extract (RTE) and its isolated compounds Ajmaline (RTC1) and Reserpine (RTC2) on triple-negative breast cancer cell line (MDA-MB-231) focusing on anti-proliferative effects. Our study imparts that RTE and RTC2 showed promising cytotoxic effects compared to RTC1. So further experiments have proceeded with RTE and RTC2, to evaluate its proliferation, migration, and apoptotic effect. The result shows around 80% of cells were observed in the G0/G1 phase in cell cycle analysis indicating the cell cycle inhibition and duel staining clearly showed the apoptotic effect. The migration of cells after the scratch was 60.45% observed in control and 90% in treated cells showing the inhibition of migration. ROS distribution was intense compared to control indicating the increased ROS stress in treated cells. Both RTE and RTC2-treated cells showed the potential to suppress proliferation and induce apoptotic change by upregulating BAX and MST-1 and suppressing Bcl2, LATS-1, and YAP, proving that deregulation of YAP resulting in the blockage of TEAD-YAP complex and inhibit proliferation. Therefore, R. tetraphylla extract and its isolated compounds were demonstrated to find its ability to act against MDA-MB-231 and these findings will help adjudicate it as a therapeutic drug against experimental triple-negative breast cancer.

Proxy-approach in understanding the bisubstrate activity of strictosidine synthases.

Besides tryptamine (1) and secologanin (2), non-cognate substrates also undergo a Pictet-Spengler reaction (PSR) catalyzed by strictosidine synthases (STR) with differing catalytic properties. We characterized the bisubstrate binding aspect of catalysis - order, affinity, and cooperativity - with STR orthologs from Rauvolfia serpentina (RsSTR) and Ophiorrhiza pumila (OpSTR) by an isothermal titration calorimetry (ITC) based 'proxy approach' that employed a non-reactive tryptamine analog (m1) to capture its inert ternary complexes with STRs and (2). ITC studies with OpSTR and (2) revealed 'tryptamine-first' cooperative binding with (1) and a simultaneous cooperative binding with (m1). Binding cooperativity among (m1) and (2) towards OpSTR was higher than RsSTR. Crystallographic study of RsSTR-(m1) complex helped to understand the unreactive binding of (m1) in terms of orientation and interactions in the RsSTR pocket. PSR with (m1) was revealed to be energetically unfeasible by the density functional theory (DFT) scans of the first hydrogen abstraction by RsSTR. The effect of pH on the bisubstrate binding to OpSTR was deciphered by molecular dynamics simulations (MDS), which also provided a molecular basis for the stability of complex of OpSTR with (m1) and (2). Therefore, we investigated STRs from a substrate binding perspective to inform drug-design and rational enzyme engineering efforts.

Alkaloids and Lignans from the Aerial Parts of Rauvolfia tetraphylla Inhibit NO Production in LPS-activated RAW 264.7 Cells.

Three previously undescribed compounds named rauvolphyllas A-C (1-3), along with thirteen known compounds, 18ß-hydroxy-3-epi-α-yohimbine (4), yohimbine (5), α-yohimbine (6), 17-epi-α-yohimbine (7), (E)-vallesiachotamine (8), (Z)-vallesiachotamine (9), 16S-E-isositsirikine (10), Nb -methylisoajimaline (11), Nb -methylajimaline (12), ajimaline (13), (+)-lyoniresinol 3α-O-ß-D-glucopyranoside (14), (+)-isolarisiresinol 3α-O-ß-D-glucopyranoside (15), and (-)-lyoniresinol 3α-O-ß-D-glucopyranoside (16) were isolated from the aerial parts of Rauvolfia tetraphylla L. Their chemical structures were elucidated based on the extensive spectroscopic interpretation of HR-ESI-MS, 1D and 2D NMR spectra. The absolute configurations of 2 and 3 were determined by experimental ECD spectra. Compounds 5, 6, 7, and 11-13 exhibited nitric oxide production inhibition activity in LPS-activated RAW 264.7 cells with the IC50 values of 79.10, 44.34, 51.28, 33.54, 37.67, and 28.56 µM, respectively, compared to that of the positive control, dexamethasone, which showed IC50 value of 13.66 µM. The other isolates were inactive with IC50 values over 100 µM.

Green synthesis of MnO2-embedded Rauvolfia tetraphylla leaves (MnO2@RTL) for crystal violet dye removal and as an antibacterial agent.

The application of green synthesized nanocomposites for the prevention of environmental pollution is increasing nowadays. Here, a green composite has been synthesized by embedding MnO2 on Rauvolfia tetraphylla leaves using its leaf extract hereinafter termed as MnO2@RTL, and demonstrated for crystal violet (CV) dye removal from simulated and real wastewater. The surface properties of the material were determined by scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDX), Fourier transform infrared spectra (FTIR), X-ray diffraction (XRD), and Brunauer-Emmet-Teller (BET) surface area, pHZPC, and zeta potential. The material exhibits a remarkable adsorption capacity of 61.162 mg/g at 328 K and pH 7. The adsorption was best fitted with Pseudo-second-order kinetic (R2 = 0.998) and a combination of Langmuir and Freundlich isotherm model (R2 = 0.994-0.999). The thermodynamic study revealed spontaneous (ΔG values = - 2.988 to - 4.978 kJ/mol) and endothermic (ΔH values = 6.830 to 11.018 kJ/mol) adsorption. After adsorption, 80% regeneration occurred with 50% methanol, and recycled up to five times. Advantageously, the material was able to remove CV dye in the presence of coexistent ions and from industrial wastewater, confirming field applicability. The adsorption capacity of the material is superior to previously reported materials. The standard deviation and relative standard deviations have been evaluated to be 0.000422-0.000667 and 0.473-0.749%, which suggests the reliability of the experiments. The exhausted material, after recycling, was pyrolyzed to overcome the disposal problem. It was established as a secondary adsorbent with 73% efficiency which makes the material win-win. The material showed antibacterial properties with Staphylococcus aureus bacteria with a zone of inhibition 5 mm.

The Rauvolfia tetraphylla genome suggests multiple distinct biosynthetic routes for yohimbane monoterpene indole alkaloids.

Monoterpene indole alkaloids (MIAs) are a structurally diverse family of specialized metabolites mainly produced in Gentianales to cope with environmental challenges. Due to their pharmacological properties, the biosynthetic modalities of several MIA types have been elucidated but not that of the yohimbanes. Here, we combine metabolomics, proteomics, transcriptomics and genome sequencing of Rauvolfia tetraphylla with machine learning to discover the unexpected multiple actors of this natural product synthesis. We identify a medium chain dehydrogenase/reductase (MDR) that produces a mixture of four diastereomers of yohimbanes including the well-known yohimbine and rauwolscine. In addition to this multifunctional yohimbane synthase (YOS), an MDR synthesizing mainly heteroyohimbanes and the short chain dehydrogenase vitrosamine synthase also display a yohimbane synthase side activity. Lastly, we establish that the combination of geissoschizine synthase with at least three other MDRs also produces a yohimbane mixture thus shedding light on the complex mechanisms evolved for the synthesis of these plant bioactives.

Characterization and in-depth genome analysis of a halotolerant probiotic bacterium Paenibacillus sp. S-12, a multifarious bacterium isolated from Rauvolfia serpentina.

BACKGROUND: Members of Paenibacillus genus from diverse habitats have attracted great attention due to their multifarious properties. Considering that members of this genus are mostly free-living in soil, we characterized the genome of a halotolerant environmental isolate belonging to the genus Paenibacillus. The genome mining unravelled the presence of CAZymes, probiotic, and stress-protected genes that suggested strain S-12 for industrial and agricultural purposes. RESULTS: Molecular identification by 16 S rRNA gene sequencing showed its closest match to other Paenibacillus species. The complete genome size of S-12 was 5.69 Mb, with a GC-content 46.5%. The genome analysis of S-12 unravelled the presence of an open reading frame (ORF) encoding the functions related to environmental stress tolerance, adhesion processes, multidrug efflux systems, and heavy metal resistance. Genome annotation identified the various genes for chemotaxis, flagellar motility, and biofilm production, illustrating its strong colonization ability. CONCLUSION: The current findings provides the in-depth investigation of a probiotic Paenibacillus bacterium that possessed various genome features that enable the bacterium to survive under diverse conditions. The strain shows the strong ability for probiotic application purposes.

Anti-fouling potential and in-silico analysis of carotenoid and fatty acids from Rauvolfia tetraphylla L.

Study investigated the antifouling potential ofRauvolfia tetraphyllaL. fruit, leaf and stem extracts against the marine fouling organisms throughin-vitroand in-silicoapproach. Methanolic crude extract of R. tetraphylla L.leaf exhibited maximum antibacterial potential against six fouling organisms isolated from Parangipettai coast and was further taken up for column fractionation. Twenty-four fractions were obtained, among which five fractions showed inhibitory efficiency against microfoulers of Bacillus megaterium. The active compounds present in the bioactive fraction were identified by FTIR, GC-MS and NMR (13C; 1H). The bioactive compounds that exhibited maximum antifouling activity were identified as Lycopersene (80%), Hexadecanoic acid; 1, 2-Benzenedicarboxylic acid, dioctyl ester; Heptadecene - (8) - carbonic acid - (1) and Oleic acid. Molecular docking studies of the potent anti-fouling compounds Lycopersene, Hexadecanoic acid, 1, 2-Benzenedicarboxylic acid, dioctyl ester and Oleic acid showed the binding energy of 6.6, - 3.8, -5.3 and -5.9 (Kcal/mol) and hence these compounds will act as a potential biocide to control the aquatic foulers. Moreover, further studies need to carry out in terms of toxicity, field assessment and clinical trial in order to take these biocides for a patent.

Developments in biotechnological tools and techniques for production of reserpine.

In the quest for novel medications, researchers have kept on studying nature to unearth beneficial plant species with medicinal qualities that may cure various diseases and disorders. These medicinal plants produce different bioactive secondary metabolites with immense therapeutic importance. One such valuable secondary metabolite, reserpine (C33H40N2O9), has been used for centuries to cure various ailments like hypertension, cardiovascular diseases, neurological diseases, breast cancer, and human promyelocytic leukaemia. Rauvolfia spp. (family Apocynaceae) is an essential reservoir of this reserpine. The current review thoroughly covers different non-conventional or in vitro-mediated biotechnological methods adopted for pilot-scale as well as large-scale production of reserpine from Rauvolfia spp., including techniques like multiple shoot culture, callus culture, cell suspension culture, precursor feeding, elicitation, synthetic seed production, scale-up via bioreactor, and hairy root culture. This review further analyses the unexplored and cutting-edge biotechnological tools and techniques to alleviate reserpine production. KEY POINTS: • Reserpine, a vital indole alkaloid from Rauvolfia spp., has been used for centuries to cure several ailments. • Overview of biosynthetic pathways and biotechnological applications for enhanced production of reserpine. • Probes the research gaps and proposes novel alternative techniques to meet the pharmaceutical industry's need for reserpine while reducing the over-exploitation of natural resources.

Chemical composition, antioxidant, and enzyme inhibitory properties of Rauwolfia vomitoria extract.

OBJECTIVES: Rauwolfia vomitoria is one ethno-botanicals in Nigeria used by traditional health practitioners in managing several human diseases. However, necessary information regarding its effect on enzymes implicated in the development and progression of erectile dysfunction is missing in the literature. Thus, this study investigated the antioxidant property and impact of Rauwolfia vomitoria extract on erectile dysfunction-related enzymes in vitro. METHODS: High performance liquid chromatography was used to identify and quantify Rauwolfia vomitoria's phenolic components. Then, utilizing common antioxidant assays, the extract's antioxidant properties were evaluated and finally the effect of the extract on some enzymes (AChE, arginase and ACE) implicated in erectile dysfunction was investigated in vitro. RESULTS: The results showed that the extract inhibited AChE (IC50=388.72 µg/mL), arginase (IC50=40.06 µg/mL) and ACE (IC50=108.64 µg/mL) activities. In addition, phenolic rich extract of Rauvolfia vomitoria scavenged radicals and chelated Fe2+ in concentration dependent manner. Furthermore, rutin, chlorogenic acid, gallic acid, and kaempferol were found in large quantities by HPLC analysis. CONCLUSIONS: Therefore, one of the potential reasons driving Rauwolfia vomitoria's use in folk medicine for the treatment of erectile dysfunction could be its antioxidant and inhibitory activities on several enzymes linked to erectile dysfunction in vitro.

Spatial localization of monoterpenoid indole alkaloids in Rauvolfia tetraphylla by high resolution mass spectrometry imaging.

Monoterpenoid indole alkaloids (MIAs) are a large group of biosynthetic compounds, which have pharmacological properties. One of these MIAs, reserpine, was discovered in the 1950s and has shown properties as an anti-hypertension and anti-microbial agent. Reserpine was found to be produced in various plant species within the genus of Rauvolfia. However, even though its presence is well known, it is still unknown in which tissues Rauvolfia produce reserpine and where the individual steps in the biosynthetic pathway take place. In this study, we explore how matrix assisted laser desorption ionization (MALDI) and desorption electrospray ionization (DESI) mass spectrometry imaging (MSI) can be used in the investigation of a proposed biosynthetic pathway by localizing reserpine and the theoretical intermediates of it. The results show that ions corresponding to intermediates of reserpine were localized in several of the major parts of Rauvolfia tetraphylla when analyzed by MALDI- and DESI-MSI. In stem tissue, reserpine and many of the intermediates were found compartmentalized in the xylem. For most samples, reserpine itself was mainly found in the outer layers of the sample, suggesting it may function as a defense compound. To further confirm the place of the different metabolites in the reserpine biosynthetic pathway, roots and leaves of R. tetraphylla were fed a stable-isotope labelled version of the precursor tryptamine. Subsequently, several of the proposed intermediates were detected in the normal version as well as in the isotope labelled versions, confirming that they were synthesized in planta from tryptamine. In this experiment, a potential novel dimeric MIA was discovered in leaf tissue of R. tetraphylla. The study constitutes to date the most comprehensive spatial mapping of metabolites in the R. tetraphylla plant. In addition, the article also contains new illustrations of the anatomy of R. tetraphylla.

Indole Alkaloids of Rauvolfia ligustrina and Their Anxiolytic Effects in Adult Zebrafish.

Rauvolfia species are well known as producers of bioactive monoterpene indole alkaloids, which exhibit a broad spectrum of biological activities. A new vobasine-sarpagan-type bisindole alkaloid (1: ) along with six known monomeric indoles (2, 3/4, 5: , and 6/7: ) were isolated from the ethanol extract of the roots of Rauvolfia ligustrina. The structure of the new compound was elucidated by interpretation of their spectroscopic data (1D and 2D NMR and HRESIMS) and comparison with published data for analog compounds. The cytotoxicity of the isolated compounds was screened in a zebrafish (Danio rerio) model. The possible GABAergic (diazepam as the positive control) and serotoninergic (fluoxetine as the positive control) mechanisms of action in adult zebrafish were also evaluated. No compounds were cytotoxic. Compound 2: and the epimers 3: /4: and 6: /7: showed a mechanism action by GABAA, while compound 1: showed a mechanism action by a serotonin receptor (anxiolytic activity). Molecular docking studies showed that compounds 2: and 5: have a greater affinity by the GABAA receptor when compared with diazepam, whereas 1: showed the best affinity for the 5HT2AR channel when compared to risperidone.

Pectic polysaccharides derived from Hainan Rauwolfia ameliorate NLR family pyrin domain-containing 3-mediated colonic epithelial cell pyroptosis in ulcerative colitis.

Pectic polysaccharides (PPs) could exert functions on ulcerative colitis (UC), which is classified as a nonspecific inflammatory disorder. This study investigated the molecular mechanism of PPs derived from Rauwolfia in UC. First, the dextran sodium sulfate (DSS)-induced mouse colitis models and lipopolysaccharide (LPS)-treated colonic epithelial cell (YAMC) models were established and treated with PP. Subsequently, the effects of PPs on mucosal damages in DSS mice were detected, and the levels of inflammatory cytokines, pyroptosis-related factors, oxidative stress-related markers, and the tight junction-related proteins in the tissues or cells were examined, and the results suggested that PPs ameliorated colonic mucosal damages and cell pyroptosis in DSS mice, and limited colonic epithelial cell pyroptosis in in vitro UC models. Subsequently, the binding relations of retinol-binding protein 4 (RBP4) to miR-124-3p and NLR pyrin domain-containing 3 (NLRP3) were analyzed. miR-124-3p targeted RBP4 and reduced the binding of RBP4 to NLRP3, thus inhibiting NLRP3-mediated pyroptosis. Finally, functional rescue experiments revealed that miR-124-3p suppression or RBP4 overexpression promoted colonic epithelial cell pyroptosis. Collectively, Rauwolfia-derived PPs limited miR-124-3p and targeted RBP4 and reduced the binding potency of RBP4 to NLRP3 to inhibit NLRP3-mediated pyroptosis, resulting in the alleviation of colonic epithelial cell pyroptosis and mucosal damages in UC.

Two new antimicrobial monoterpenoid indole alkaloids from the roots of Rauvolfia yunnanensis.

Two new monoterpenoid indole alkaloids 3-hydroxylochnerine (1) and 10-hydroxyvinorine (2) were isolated from the roots of Rauvolfia yunnanensis. Their structures were elucidated based on the analysis of spectroscopic data and ECD calculation. Both compounds exhibited potent antimicrobial activity against Bacillus subtilis and Escherichia coli, and their activities were comparable to the well-known antibacterial drug berberine.

[Rauwolfia extract inhibits the proliferation of prostate cells in rats with benign prostatic hyperplasia].

OBJECTIVE: To investigate the effects of different concentrations of Rauwolfia extract (RE) on the proliferation of prostate cells in the rat model of benign prostatic hyperplasia (BPH). METHODS: We randomly divided 48 male SD rats into six groups of an equal number, BPH model control, finasteride, low-concentration RE, medium-concentration RE, high-concentration RE and normal control, and established a BPH model in the former five groups by subcutaneous injection of testosterone propionate following castration. We treated the rats of the finasteride and RE groups intragastrically with finasteride solution at 5 mg/kg and RE at 5, 10 and 20 mg/kg respectively, and those of the model control and normal control groups with an equal dose of normal saline, all once a day for 28 consecutive days. Then, we killed all the animals, collected their prostate tissue, obtained the wet weight and volume of the prostate, the prostate index and the contents of serum T and dihydrotestosterone (DHT), observed the morphological changes of the prostate tissue by HE staining, counted the glands in the prostate tissue, measured the intraglandular area, and determined the expressions of PCNA and α-SMA by immunohistochemistry. RESULTS: Compared with the rats of the normal control group, the BPH model controls showed significantly increased wet weight (ï¼»0.923 ± 0.15ï¼½ vs ï¼»1.455 ± 0.52ï¼½ g, P < 0.05), volume (ï¼»1.035 ± 0.29ï¼½ vs ï¼»1.687 ± 0.31ï¼½ ml, P < 0.05) and index of the prostate (ï¼»0.23 ± 0.04ï¼½% vs ï¼»0.37 ± 0.15ï¼½%, P < 0.05), dilation, hyperemia and edema of the prostatic stroma and vessels, and proliferation rate of the prostatic cells, but remarkably decreased number of glands (ï¼»20.35 ± 3.83ï¼½ vs ï¼»12.56 ± 2.58ï¼½, P < 0.05), epithelial thickness (ï¼»39.76 ± 5.20ï¼½ vs ï¼»19.52 ± 1.52ï¼½ µm, P < 0.05) and intraglandular area (ï¼»12.3 ± 1.21ï¼½ vs ï¼»5.96 ± 0.34ï¼½ ×103µm2, P < 0.05). In comparison with the BPH model controls, the animals treated with RE, especially in the high-concentration RE group, exhibited marked decreases in the weight (ï¼»1.455 ± 0.52ï¼½ vs ï¼»0.862 ± 0.31ï¼½ g, P < 0.05), volume ( ï¼»1.687 ± 0.31ï¼½ vs ï¼»0.952 ± 0.28ï¼½ ml, P < 0.05) and index of the prostate (ï¼»0.37 ± 0.15ï¼½% vs ï¼»0.22 ± 0.07ï¼½%, P < 0.05), dramatic improvement in the number of glands (ï¼»12.56 ± 2.58ï¼½ vs ï¼»18.36 ± 1.25ï¼½, P < 0.05), epithelial thickness (ï¼»39.76 ± 5.20ï¼½ vs ï¼»19.04 ± 3.89ï¼½ µm, P < 0.05) and intraglandular area (ï¼»5.96 ± 0.34ï¼½ vs ï¼»10.25 ± 0.98ï¼½ ×103µm2, P<0.05ï¼½, P < 0.05), remarkable down-regulation of the expressions of PCNA and α-SMA, and significant reduction of the contents of serum T (ï¼»19.147 ± 3.214ï¼½ vs ï¼»6.016 ± 1.978ï¼½ ng/ml, P < 0.05) and DHT (ï¼»9.052 ± 0.633ï¼½ vs ï¼»2.532 ± 0.386ï¼½ ng/ml, P < 0.05). CONCLUSION: Rauwolfia extract can inhibit the proliferation of prostate cells and relieve BPH symptoms in a concentration-dependent manner in rats with BPH.

Extracts of the Medicinal Plants Pao Pereira and Rauwolfia vomitoria Inhibit Ovarian Cancer Stem Cells In Vitro.

Ovarian cancer has an enrichment of cancer stem cells (CSCs) which contribute to the treatment resistant tumor's high rate of recurrence and metastasis. Here we investigated 2 plant extracts from the medicinal plants Pao Pereira (Pao) and Rauwolfia vomitoria (Rau) each for their activities against ovarian CSCs. Both Pao and Rau inhibited overall proliferation of human ovarian cancer cell lines with IC50 ranging from 210 to 420 µg/mL and had limited cytotoxicity to normal epithelial cells. Ovarian CSC population was examined using cell surface markers and tumor spheroid formation assays. The results showed that both Pao and Rau treatment significantly reduced the ovarian CSC population. Pao and Rau had similar activities in inhibiting ovarian CSCs, with IC50s of ~120 µg/mL for 24 hours treatment, and ~50 µg/mL for long-term tumor spheroid formation. Nuclear ß-catenin levels were decreased, suggesting suppression of Wnt/ß-catenin signaling pathway. Taken together, data here showed that Pao and Rau both inhibited ovarian cancer stem cells, probably in preference to the bulk of tumor cells. Further mechanistic studies and in vivo investigation validating these findings are warranted, given that inhibition of cancer stem cells holds the promise of comprehensively inhibiting cancer metastasis, drug resistance and recurrence.

A serum metabolomics study based on LC-MS: Chemosensitization effects of Rauvolfia vomitoria Afzel. combined with 5- fluorouracil on colorectal cancer mice.

Colorectal cancer (CRC) is one of the malignant tumors with high incidence, and is mainly treated by chemotherapy at present. However, during CRC treatment, long-term use of traditional chemotherapeutic drugs will reduce the sensitivity of chemotherapy. Our previous studies have shown that Rauvolfia vomitoria total alkaloids (RVA) played an important role in 5-fluorouracil (5-FU) chemosensitization in CRC therapy, but its intervention mechanism has not been clarified completely in the metabolic level. Therefore, in this study, LC-MS based metabolomics was employed to explore the mechanism of 5-FU chemosensitization in CRC induced by the combination of RVA and conventional chemotherapeutic with 5-FU. The results showed that the final tumor weight of the high-dose combined group was significantly different from that of the 5-FU alone group. To evaluate the chemosensitization effects of RVA, serum samples collected from six groups (six mice in each group) with different administration methods were analyzed by HPLC-Q-Exactive Orbitrap/MS. After multivariate statistical analysis and metabolites identification, 25 different metabolites were identified between the 5-FU treatment group and combined high-dose treatment group, among which lipid and fatty acid metabolism pathways were mostly affected. These results suggest that RVA may sensitize traditional chemotherapeutic drug 5-FU and exert anti-tumor activity through influencing lipid metabolism and cell energy metabolism. Metabolomics provided a new insight into estimate of the therapeutic effect and dissection of the potential mechanisms of traditional Chinese medicine in treating colorectal cancer.

Biotechnological interventions and indole alkaloid production in Rauvolfia serpentina.

Rauvolfia serpentina (L). Benth. ex Kurz. (Apocynaceae), commonly known as Sarpagandha or Indian snakeroot, has long been used in the traditional treatment of snakebites, hypertension, and mental illness. The plant is known to produce an array of indole alkaloids such as reserpine, ajmaline, amalicine, etc. which show immense pharmacological and biomedical significance. However, owing to its poor seed viability, lesser germination rate and overexploitation for several decades for its commercially important bioactive constituents, the plant has become endangered in its natural habitat. The present review comprehensively encompasses the various biotechnological tools employed in this endangered Ayurvedic plant for its in vitro propagation, role of plant growth regulators and additives in direct and indirect regeneration, somatic embryogenesis and synthetic seed production, secondary metabolite production in vitro, and assessment of clonal fidelity using molecular markers and genetic transformation. In addition, elicitation and other methods of optimization of its indole-alkaloids are also described herewith. KEY POINTS: • Latest literature on in vitro propagation of Rauvolfia serpentina • Biotechnological production and optimization of indole alkaloids • Clonal fidelity and transgenic studies in R. serpentina.

Rauwolfia vomitoria extract suppresses benign prostatic hyperplasia by inducing autophagic apoptosis through endoplasmic reticulum stress.

BACKGROUND: The current drug treatments for benign prostatic hyperplasia (BPH) have negative side effects. Therefore, it is important to find effective alternative therapies with significantly fewer side effects. Our previous study revealed that Rauwolfia vomitoria (RWF) root bark extract reversed BPH development in a rat model. However, the molecular mechanism of its inhibitory effects on BPH remains largely unknown. METHODS: BPH-1 and WPMY-1 cell lines derived from BPH epithelial and prostatic stromal compartments were selected to investigate how RWF extract inhibits BPH in vitro by MTT and flow cytometry assays. Microarray, quantitative real-time PCR, immunoblotting, and GFP-LC3 immunofluorescence assays were performed to evaluate the effects of RWF extract on endoplasmic reticulum stress (ER stress) and autophagic apoptosis pathways in two cell lines. A human BPH ex vivo explant assay was also employed for validation. RESULTS: RWF extract treatment decreased cell viability and induced apoptotic cell death in both BPH-1 and WPMY-1 cells in a concentration-dependent manner with the increase of pro-apoptotic PCDC4 protein. RWF extract induced autophagy by enhancing the levels of autophagic genes (ULK2 and SQSTM1/p62) and the LC3II:LC3I ratio, with the increase of GFP-LC3 puncta. Moreover, RWF extract activated PERK- and ATF6-associated ER stress pathways by inducing the transcriptional levels of EIF2AK3/PERK, DDIT3/CHOP and ATF6, accompanied by the reduction of BiP protein level, but not its mRNA level. Another ER stress pathway was not induced by RWF extract, as manifested by the lack of XBP1 splicing. Pharmacological inhibition of autophagy by 3-methyladenine abrogated apoptosis but not ER stress; while inhibition of ER stress by 4-phenylbutyrate alleviated the induction of autophagy and apoptosis. In addition, pretreatments with either 3-methyladenine or 4-phenylbutyrate suppressed RWF extract-induced cytotoxicity. Notably, the inductions of PERK- and ATF6-related stress pathways and autophagic apoptosis were confirmed in a human BPH ex vivo explant. CONCLUSIONS: Our data have demonstrated that RWF extract significantly suppressed the viabilities of BPH epithelial cells and BPH myofibroblasts by inducing apoptosis via upregulating ER stress and autophagy. These data indicate that RWF extract is a potential novel alternative therapeutic approach for BPH.