Mirabilis Himalaica Research Articles

Integration of Metabolomics with Network Pharmacology Deciphers the Anti-Fatigue Activity Mechanisms of the Extract of Mirabilis himalaica Root.

Fatigue, a common symptom in both diseased and healthy individuals, is a biological phenomenon characterized by a sense of extreme physical or mental exhaustion. To explore novel drugs and food sources of anti-fatigue, the hydroalcoholic extract of the root of Mirabilis himalaica (MH extract) is evaluated as anti-fatigue agents in this work, and clarifies that the mechanism of MH intervention in fatigue symptoms, and distribution of the anti-fatigue constituents in the plant of Mirabilis himalaica is examined. The results show that the MH extract have a significantly anti-fatigue effect via the pharmacological experiment and biochemical indicators. The network pharmacology, metabolomics, molecular docking, and pharmacology are integrated to determine that boeravinone A, B, and E are the pharmacoperones of anti-fatigue. Moreover, the compounds of boeravinone are present only in the root and not in the leaf and stem of the Mirabilis himalaica, which validates that root of Mirabilis himalaica is historically and officially utilized medicinal parts.

Endophytic bacterial community structure and diversity of the medicinal plant Mirabilis himalaica from different locations

Endophytic bacteria play important roles in medicinal plant growth, abiotic stress, and metabolism. Mirabilis himalaica (Edgew.) Heimerl is known for its medicinal value as Tibetan traditional plant; however, little is known about the endophytic bacteria associated with this plant in different geographic conditions and vegetal tissues. To compare the endophytic bacterial community associated with this plant in different geographic conditions and vegetal tissues, we collected the leaves, stems, and roots of M. himalaica from five locations, Nongmu college (NM), Gongbujiangda (GB), Zhanang County (ZL), Lang County (LX), and Sangri County (SR), and sequenced the 16S rRNA V5-V7 region with the Illumina sequencing method. A total of 522,450 high-quality sequences and 4970 operational taxonomic units (OTUs) were obtained. The different tissues from different locations harbored unique bacterial assemblages. Proteobacteria and Actinobacteria were the dominant phyla in all the samples, while the dominant genera changed based on the different tissues. The endophytic bacterial structures in the leaf and stem tissues were different compared to root tissues. Redundancy analysis (RDA) showed that the endophytic bacterial community was significantly correlated with pH, available phosphorus (AP), total phosphorus (TP), total nitrogen (TN), and soil organic matter (SOM). These findings suggested that the geographic conditions, climate type, ecosystem type, and tissues determined the endophytic bacterial composition and relative abundances. This conclusion could facilitate an understanding of the relationship and ecological function of the endophytic bacteria associated with M. himalaica and provide valuable information for artificial planting of M. himalaica and identifying and applying functional endophytic bacteria.

Insights into the mechanism underlying UV-B induced flavonoid metabolism in callus of a Tibetan medicinal plant Mirabilis himalaica

Mirabilis himalaica is an important Tibetan medicinal plant in China. However, it has become a rare and class I endangered Tibetan medicine plant. Therefore, the use of callus to propagate germplasm resources is of great significance. We found that the flavonoid content of M. himalaica callus increased continuously with the extension of UV-B treatment. Multi-omics profiles were used to reveal the co-expression patterns of gene networks of flavonoid metabolism in M. himalaica callus during UV-B radiation. Results showed that five medicinal metabolics, including geranin, eriodictyol, astragalin, isoquercetin, pyrotechnic acid, and one anthocyanin malvide-3-O-glucoside were identified. The transcriptome data were divided into 46 modules according to the expression pattern by WGCNA (weighted gene co-expression network analysis), of which the module Turquoise had the strongest correlation with six target metabolites. We found that seven structural genes and twenty-five transcription factors were related to the metabolism of flavonoid synthesis, among which the structural genes CHI, C4H and UGT79B6 had strong co-expression relationships with the 6 target metabolites. WRKY42, WRKY7, bHLH128 and other transcription factors had strong co-expression relationships with multiple structural genes. Consequently, these findings suggest callus grown under UV-B treatment could be an effective alternative medical resource of M. himalaica, which is valuable for conservation and usage of this wild and endangered plant.

Staying in situ or shifting range under ongoing climate change: A case of an endemic herb in theHimalaya‐HengduanMountains across elevational gradients

AbstractAimHow species respond to ongoing climate change has been a hot research topic, especially with the controversy in shifting range (movement) or persisting in local habitat (in situ) as the primary response. Assessing the relative roles of range shifts, phenotypic plasticity and genetic adaptation helps us predict the evolutionary fate of species. We aim to explore the evolutionary strategies of plants under climate change from a keystone herb in alpine ecosystems,Mirabilis himalaica, along its elevational gradient.LocationHimalaya‐Hengduan Mountains, China.MethodsWe combined evidence from population genomics and ecological data in both space and time to investigate the state of “staying” or “moving”. We identified migration events by assessing historical and contemporary gene flow and changes in species distribution. Morphological variation was compared by measuring five traits using specimen data. Moreover, we explored climate‐driven genetic variation and local selection regimes acting on populations in the alpine landscape along an elevational gradient.ResultsOur results argue that staying in situ by morphological variation and local genetic evolution rather than range shifting plays an important role inM. himalaicaresponse to climate change. We first found trace evidence of upward or climatic‐driven shifting along an elevational gradient, although asymmetric gene flow was restricted within microenvironments of mid‐elevational populations. Furthermore, morphological variation comparisons revealed clinal variation, as resource allocation showed a declining pattern in vegetative growth but increased reproductive growth with increasing elevation. Outlier tests and environment association analyses indicated adaptative loci primarily related to thermal‐driven selection and continuous adaptations to high elevation in the Himalaya‐Hengduan Mountains.Main ConclusionsOur findings showM. himalaicamay persist in local habitats rather than shifting range under climate change, exhibiting a low risk of genomic vulnerability in current habitats. This study has important implications in improving our understanding of the evolutionary response in alpine plants to climate change.

Physicochemical properties, immunostimulatory and antioxidant activities of a novel polysaccharide isolated from Mirabilis himalaica (Edgew) Heim.

Herbal medicines often contain bioactive polysaccharides. However, many medicinal herbs have not been explored for any active saccharides that may play key roles in their bioactivities. Herein, we extracted a novel polysaccharide from Mirabilis himalaica (Edgew) heim (denoted MHHP), a popular medicinal ingredient in traditional medicines. The structural and morphological characteristics of MHHP were measured and elucidated by high-performance gel permeation chromatography, gas chromatography connected with mass spectrometry, Fourier transform infrared and nuclear magnetic resonance spectroscopy as well as scanning electron microscopy. MHHP was homogeneous with a molecular weight of 16.1 kDa, Mw/Mn = 1.33, containing mainly α-d-glucan residues with (1→4)-linkage. The biological activities of MHHP upon proliferation of splenic lymphocyte, activation of related cytokine and production of nitric oxide (NO) in RAW264.7 cells were investigated in vitro. MHHP induced proliferation of mouse spleen lymphocytes and significantly promoted the secretion in TNF-α, IL-6 and NO production in RAW264.7 cells. Meanwhile, MHHP exhibited relatively low antioxidant abilities. Our data suggested that MHHP may have potential immunoregulatory and anti-inflammatory activity, with a moderate antioxidant activity.

Achene mucilage formation process and extrusion from hydrated pericarp of Mirabilis himalaica

Myxospermy is an important feature of achenes of the alpine plant Mirabilis himalaica, and the achene mucilage increases the germination rate and early seedling growth during exposure to abiotic stresses, which has important functions that allow M. himalaica to survive the extreme climate of the Tibet Plateau. However, achene formation and mucilage extrusion are poorly understood. In the present study, comprehensive analyses were performed on mucilage production during achene development and mucilage release from hydrated achene pericarp in M. himalaica. First, fertilization initiated the development of M. himalaica achenes, during which their color, size and texture were altered dramatically. Second, using a metachromatic staining procedure, cytological events, the establishment of mucilage secretory cells in the inner epicarp layer were observed. The hydration of mature achenes led to the rapid bursting of mucilage secretory cells, which released a hydrophilic gel that surrounded the achenes. Finally, enzymatic digestion indicated that major components of the mucilage were pectins; glucose (41.40%), rhamnose (26.58%), galactose (18.33%), trehalose (12.12%), and mannose (1.57%) were found to be the components of achene by using ion-exchange chromatography.

Characterization of the complete chloroplast genome sequence of the medicinal plant Mirabilis himalaica

Mirabilis himalaica is an old and popular medicinal plant used in traditional Tibetan folk medicine. Here, we reported the complete chloroplast genome sequence of Mirabilis himalaica. The assembled chloroplast genome was 154,348 bp long, containing a large single-copy region of 85,809 bp, a small single-copy region of 17,935 bp, and a pair of inverted repeat regions of 25,302 bp. It had 36% GC content and encoded 131 genes including 86 protein-coding genes, eight rRNA genes, and 37 tRNA. Fifteen and two genes contained one and two introns, respectively. Phylogenetic analysis revealed that Mirabilis himalaica was sister to Nyctaginia capitata.

The complete mitochondrial genome sequence of Mirabilis himalaica, an endemic plant to Tibet

The complete mitochondrial genome of the important medicinal plant Mirabilis himalaica has been reported in this study. It was 346,363 bp in size with a GC content of 44.6% and contained 42 protein-coding genes (30 known functional genes, 11 ORF genes and one ycf2 gene of unknown function), 27 tRNA genes, and four rRNA genes. Most of the genes were single-copy genes, only three were duplicated and two were multi-copy. Phylogenetic analysis showed M. himalaica to be closely clustered with Silene latifolia, Spinacia oleracea and Beta vulgaris, all species belong to the order Caryophyllales.

Geological and Climatic Factors Affect the Population Genetic Connectivity in Mirabilis himalaica (Nyctaginaceae): Insight From Phylogeography and Dispersal Corridors in the Himalaya-Hengduan Biodiversity Hotspot.

The genetic architecture within a species in the Himalaya-Hengduan Mountains (HHM) region was considered as the consolidated consequence of historical orogenesis and climatic oscillations. The visualization of dispersal corridors as the function of population genetic connectivity became crucial to elucidate the spatiotemporal dynamics of organisms. However, geodiversity and physical barriers created by paleo geo-climatic events acted vigorously to impact notable alterations in the phylogeographic pattern and dispersal corridors. Therefore, to achieve detailed phylogeography, locate dispersal corridors and estimate genetic connectivity, we integrated phylogeography with species distribution modelling and least cost path of Mirabilis himalaica (Edgew.) Heimerl in the HHM. We amplified four cpDNA regions (petL-psbE, rps16-trnK, rps16 intron, trnS-trnG), and a low copy nuclear gene (G3pdh) from 241 individuals of 29 populations. SAMOVA, genealogical relationships, and phylogenetic analysis revealed four spatially structured phylogroups for M. himalaica with the onset of diversification in late Pliocene (c. 3.64 Ma). No recent demographic growth was supported by results of neutrality tests, mismatch distribution analysis and Bayesian skyline plot. Paleo-distribution modelling revealed the range dynamics of M. himalaica to be highly sensitive to geo-climatic change with limited long-distance dispersal ability and potential evolutionary adaptation. Furthermore, river drainage systems, valleys and mountain gorges were identified as the corridors for population genetic connectivity among the populations. It is concluded that recent intense mountain uplift and subsequent climatic alterations including monsoonal changes since Pliocene or early Pleistocene formulated fragmented habitats and diverse ecology that governed the habitat connectivity, evolutionary and demographic history of M. himalaica. The integrative genetic and geospatial method would bring new implications for the evolutionary process and conservation priority of HHM endemic species.

Comprehensive Investigation of the Differences of the Roots of Wild and Cultivated Mirabilis himalaica (Edgew) Heim Based on Macroscopic and Microscopic Identification Using HPLC Fingerprint.

Mirabilishimalaica (Edgew) Heim (MH) is an important Tibetan medicine with demonstrated medicinal efficacy and promising developmental value. A previous study of MH was limited to vague morphological and microscopic descriptions, restricting its clinical application and further development as a medicine. The goal of this study was to comprehensively characterize wild and cultivated products of MH using macroscopic and microscopic identification using HPLC fingerprint. The results revealed that the cultivated and wild MH exhibited differences in macroscopic and microscopic characteristics and chemical components. This analysis can facilitate the establishment of a more comprehensive quality evaluation method for MH. These results provide the basis for clinical applications and the improvement of quality standards of MH as a step towards modernization of Tibetan medicine.

Complete chloroplast genome sequence of Mirabilis himalaica (Nyctaginaceae)

Mirabilis himalaica (Nyctaginaceae) is endemic to the Himalayas where it is used in traditional Tibetan folk medicine. In this study, we first presented the complete chloroplast genome of M. himalaica. Complete genome size of M. himalaica ranged from 154,348 to 154,388 bp. The length varied from 85,808 to 85,845 bp in the (large single-copy) LSC region, from 17,935 to 17,938 bp in the (small single-copy) SSC region, and from 25,302 to 25,303 bp in the inverted repeat (IR) region. The overall GC contents of the chloroplast genome sequences were around 36%. Annotation analysis revealed a total of 112 genes, including 78 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. The phylogenetic analysis with three M. himalaica samples and five other Nyctaginaceae species showed that Mirabilis including two species was clustered with high bootstrap support. The complete chloroplast genome sequences obtained in this study will provide valuable data for wider studies into the phylogenetics and conservation biology of M. himalaica.

Integrated Analysis of Transcriptomic and Proteomics Data Reveals the Induction Effects of Rotenoid Biosynthesis of Mirabilis himalaica Caused by UV-B Radiation.

Mirabilis himalaica (Edgew.) Heimerl is one of the most important genuine medicinal plants in Tibet, in which the special plateau habitat has been associated with its excellent medicinal quality and efficacy. However, the mechanisms by which environmental factors affect biosynthesis of secondary metabolic components remain unclear in this species. In this study, RNA sequencing and iTRAQ (isobaric Tags for Relative and Absolute Quantification) techniques were used to investigate the critical molecular “events” of rotenoid biosynthesis responding to UV-B radiation, a typical plateau ecological factor presented in native environment-grown M. himalaica plants. A total of 3641 differentially expressed genes (DEGs) and 106 differentially expressed proteins (DEPs) were identified in M. himalaica between UV-B treatment and control check (CK). Comprehensive analysis of protein and transcript data sets resulted in 14 and 7 DEGs from the plant hormone signal transduction and phosphatidylinositol signaling system pathways, respectively, being significantly enriched. The result showed that the plant hormone signal transduction and phosphatidylinositol signaling system might be the key metabolic strategy of UV-B radiation to improve the biosynthesis of rotenoid in M. himalaica. At same time, most of the DEGs were associated with auxin and calcium signaling, inferring that they might drive the downstream transmission of these signal transduction pathways. Regarding those pathways, two chalcone synthase enzymes, which play key roles in the biosynthesis of rotenoid that were thought as the representative medicinal component of M. himalaica, were significantly upregulated in UV-B radiation. This study provides a theoretical basis for further exploration of the adaptation mechanism of M. himalaica to UV-B radiation, and references for cultivation standardization.

Flavonoids from Mirabilis himalaica

Six previously undescribed flavonoids, 2S-5-methoxy-6-methyl-7,2′-dihydroxyflavanone, 5,7,2′-trihydroxy-6-methylflavone, 5,7,6′-trihydroxy-6-methylcoumaronochromone, 2,4′,6′-trihydroxy-2′-methoxy-3′-methylchalcone, 6R,11-dimethoxy-9-hydroxyrotenoid, and 6R,11-dimethoxy-9-hydroxy-10-methylrotenoid, along with eight known flavonoids, including 2S-5-methoxy-6-methyl-7,4′-dihydroxyflavanone, not previously reported as a natural product, and seven rotenoids, boeravinone A, B, D, P, F, coccineone B, and mirabijalone E, were isolated from the ethyl acetate soluble fraction of Mirabilis himalaica roots. Their structures were established by the extensive spectroscopic analysis, including HRESIMS, UV, NMR and ECD. All compounds were evaluated for their cytotoxic activities against three human cancer cell lines: A375 (melanoma), A549 (lung), and PLC (hepatoma).

Integrated analysis of transcriptomic and metabolomic data reveals critical metabolic pathways involved in rotenoid biosynthesis in the medicinal plant Mirabilis himalaica

Mirabilis himalaica (Edgew.) Heimerl is among the most important genuine medicinal plants in Tibet. However, the biosynthesis mechanisms of the active compounds in this species are unclear, severely limiting its application. To clarify the molecular biosynthesis mechanism of the key representative active compounds, specifically rotenoid, which is of special medicinal value for M. himalaica, RNA sequencing and TOF-MS technologies were used to construct transcriptomic and metabolomic libraries from the roots, stems, and leaves of M. himalaica plants collected from their natural habitat. As a result, each of the transcriptomic libraries from the different tissues was sequenced, generating more than 10 Gb of clean data ultimately assembled into 147,142 unigenes. In the three tissues, metabolomic analysis identified 522 candidate compounds, of which 170 metabolites involved in 114 metabolic pathways were mapped to the KEGG. Of these genes, 61 encoding enzymes were identified to function at key steps of the pathways related to rotenoid biosynthesis, where 14 intermediate metabolites were also located. An integrated analysis of metabolic and transcriptomic data revealed that most of the intermediate metabolites and enzymes related to rotenoid biosynthesis were synthesized in the roots, stems and leaves of M. himalaica, which suggested that the use of non-medicinal tissues to extract compounds was feasible. In addition, the CHS and CHI genes were found to play important roles in rotenoid biosynthesis, especially, since CHS might be an important rate-limiting enzyme. This study provides a hypothetical basis for the screening of new active metabolites and the metabolic engineering of rotenoid in M. himalaica.

Role of mucilage during achene germination and sprout growth of the endangered Tibetan medicinal herb Mirabilis himalaica (Nyctaginaceae) exposed to abiotic stresses

Role of mucilage during achene germination and sprout growth of the endangered Tibetan medicinal herb Mirabilis himalaica (Nyctaginaceae) exposed to abiotic stresses

A new diphenyl ether derivative from Mirabilis himalaica

One new compound, himalain A (1), together with 12 known compounds were isolated from Mirabilis himalaica. Their structures were determined by spectroscopic methods, including 2D NMR techniques, and the absolute configuration of the 1,2-diol moiety in 1 was defined through Riguera’s method. All isolated compounds were tested for their cytotoxic and antibacterial activities.

Molecular cloning and transgenic characterization of the genes encoding chalcone synthase and chalcone isomerase from the Tibetan herbal plant Mirabilis himalaica.

Mirabilis himalaica is an endangered medicinal plant species in the Tibetan Plateau. The two genes respectively encoding chalcone synthase (MhCHS) and chalcone isomerase (MhCHI) were isolated and characterized from M. himalaica. The sequence analysis revealed that the two genes were similar with their corresponding homologous genes in other plants. The tissue profiles showed that both MhCHS and MhCHI had higher expression levels in roots than in stems and leaves. Transgenic hairy root cultures respectively with overexpressing MhCHS and MhCHI were established. The genomic PCR detection confirmed the authority of transgenic hairy root lines, in which either MhCHS or MhCHI expression levels were much higher than that in non-transgenic hairy root line. Finally, the HPLC detection results demonstrated that the rotenoid contents in MhCHS/MhCHI-transformed hairy root lines were enhanced. This study provided two candidate genes that could be used to genetic engineering rotenoid biosynthesis in M. himalaica and an alternative method to produce rotenoid using transgenic hairy root cultures.

A natural phenylpropionate derivative from Mirabilis himalaica inhibits cell proliferation and induces apoptosis in HepG2 cells

Bioactivity-guided study led to the isolation of a natural phenylpropionate derivative, (E)-3-(4-hydroxy-2-methoxyphenyl)-propenoic acid 4-hydroxy-3-methoxyphenyl ester from the roots of Mirabilis himalaica. Cellular analysis showed that compound 1 specifically inhibited the cancer cell growth through the S phase arrest. Mechanistically, compound 1 was able to induce the apoptosis in HepG2 cells through mitochondrial apoptosis pathway in which Bcl-2 and p53 were required. Interestingly, the cellular phenotype of compound 1 were shown specifically in cancer cells originated from hepatocellular carcinoma (HepG2) while compromised influence by compound 1 were detected within the normal human liver cells (L-02). Consistently, the in vivo inhibitory effects of compound 1 on tumor growth were validated by the in xenograft administrated with HepG2 cells. Our results provided a novel compound which might serve as a promising candidate and shed light on the therapy of the hepatocellular carcinoma.

Analysis of photosynthetic characteristics and its influencing factors of medicinal plant Mirabilis himalaica

To study photosynthetic characteristics and its influencing factors in leaves of medicinal plant Mirabilis himalaica, and provide an evidence for guiding artificial planting and improving the quantity. The light-response and diurnal photosynthesis course of leaves at the booting stages of 1-3 year old M. himalaica were measured with LI-6400 system. The Results showed that the light response curves were fitted well by non rectangle hyperbola equation (R2 > or = 0.98). The values of the maximum photosynthetic rate (Pmax) and light use efficiency of three-year old M. himalaica leaves were higher than those of 1-2 year old individuals. The diurnal variation of net photosynthetic rate (Pn) and stomatal conductance (Gs) of 2-3 year old M. himalaica were typical double-peak curves determinately regulated by stomatal conductance. However, transpiration rate (Tr) of 1-3 year old plants leaves were single-peak curve, which was self-protection of harm reduction caused by the higher temperature at noontime. Correlation analysis showed that the changes between photosynthetic active radiation (PFD), air temperature (T ) and Pn, were significant positive related. Therefore, M. himalaica is a typical sun plant, which should be planted under the sufficient sunshine field and prolong the growing ages suitably in order to improve the yield.

Mirabijalone E: A novel rotenoid from Mirabilis himalaica inhibited A549 cell growth in vitro and in vivo

Ethnopharmacological relevanceThe roots of Mirabilis himalaica have been used in Tibetan folk medicine for treatment of uterine cancer, nephritis edematous, renal calculus and arthrodynia. In our previous work, the ethanol extract of roots had shown potent cytotoxicity against human cancer cells. However, no information is available on the antitumor effect of Mirabilis himalaica. The aim of the present study was to investigate the active constituents guided by bioassay and evaluate the related antitumor efficacy in vitro and in vivo. Materials and methodsThe active subextract (ethyl acetate) was subjected to successive chemical separation using a combination of silica gel, LH-20 chromatography and semi-preparative HPLC. The structures were determined by spectroscopic analysis techniques such as nuclear magnetic resonance (NMR) and mass spectrometry. Three human cancer cell lines, A549, HepG2 and HeLa were used for in vitro cytotoxicity evaluation of all isolated compounds by MTT-assay. Then, the potent and novel compound mirabijalone E was employed to the mechanism study againstA549 cells. BrdU immunofluorescence, soft agar assay and cell cycle analysis were employed to detect the cell proliferation effects. Annexin V-FITC/PI staining assay was used for examining apoptotic effects. Expression levels of apoptosis-related proteins were determined by western blot assay. in vivo tumorigenic assay was used to evaluate the xenograft tumor growth treated with mirabijalone E. ResultsOne new rotenoid compound, mirabijalone E, together with eight known rotenoids was isolated from Mirabilis himalaica. Mirabijalone E, 9-O-methyl-inone B, boeravinone C and boeravinone H exhibited cytotoxicity against A 549 and HeLa cells. Further study on mirabijalone E was carried out in vitro and in vivo. Mirabijalone E inhibited A549 cells growth in a time and dose-dependent manner, which arrested cell cycle in S phase. Mechanistically, mirabijalone E treatment resulted in the increase of Bax expression level, the decrease of Bcl-2 level and the activation of caspase-3, which suggested the activation of apoptosis cascades. Consequently, the xenograft treated with mirabijalone E showed markedly suppressed tumor growth. ConclusionsThe result suggested that mirabijalone E, together with active compounds, 9-O-methyl-4-hydroxyboeravinone B, boeravinone C and boeravinone H could be a promising candidate for cancer therapy.