Integrated component identification, network pharmacology, and experimental verification revealed mechanism of Dendrobium officinale Kimura et Migo against lung cancer.

BACKGROUND: Dendrobium officinale Kimura et Migo (DO), a valuable Chinese herbal medicine, has been reported to exhibit potential effects in the prevention and treatment of lung cancer. However, its material basis and mechanism of action have not been comprehensively analyzed. PURPOSE: The objective of this study was to preliminarily elucidate the active components and pharmacological mechanisms of DO in treating lung cancer, according to UPLC-Q/TOF-MS, HPAEC-PAD, network pharmacology, molecular docking, and experimental verification. METHODS: The chemical components of DO were identified via UPLC-Q/TOF-MS, while the monosaccharide composition of Dendrobium officinale polysaccharide (DOP) was determined by HPAEC-PAD. The prospective active constituents of DO as well as their respective targets were predicted in the combined database of Swiss ADME and Swiss Target Prediction. Relevant disease targets for lung cancer were searched in OMIM, TTD, and Genecards databases. Further, the active compounds and potential core targets of DO against lung cancer were found by the C-T-D network and the PPI network, respectively. The core targets were then subjected to enrichment analysis in the Metascape database. The main active compounds were molecularly docked to the core targets and visualized. Finally, the viability of A549 cells and the relative quantity of associated proteins within the major signaling pathway were detected. RESULTS: 249 ingredients were identified from DO, including 39 flavonoids, 39 bibenzyls, 50 organic acids, 8 phenanthrenes, 27 phenylpropanoids, 17 alkaloids, 17 amino acids and their derivatives, 7 monosaccharides, and 45 others. Here, 50 main active compounds with high degree values were attained through the C-T-D network, mainly consisting of bibenzyls and monosaccharides. Based on the PPI network analysis, 10 core targets were further predicted, including HSP90AA1, SRC, ESR1, CREBBP, MAPK3, AKT1, PIK3R1, PIK3CA, HIF1A, and HDAC1. The results of the enrichment analysis and molecular docking indicated a close association between the therapeutic mechanism of DO and the PI3K-Akt signaling pathway. It was confirmed that the bibenzyl extract and erianin could inhibit the multiplication of A549 cells in vitro. Furthermore, erianin was found to down-regulate the relative expressions of p-AKT and p-PI3K proteins within the PI3K-Akt signaling pathway. CONCLUSIONS: This study predicted that DO could treat lung cancer through various components, multiple targets, and diverse pathways. Bibenzyls from DO might exert anti-lung cancer activity by inhibiting cancer cell proliferation and modulating the PI3K-Akt signaling pathway. A fundamental reference for further studies and clinical therapy was given by the above data.

Medical Plant Extract Purification from Cadmium(II) Using Modified Thermoplastic Starch and Ion Exchangers.

Pure compounds extracted and purified from medical plants are crucial for preparation of the herbal products applied in many countries as drugs for the treatment of diseases all over the world. Such products should be free from toxic heavy metals; therefore, their elimination or removal in all steps of production is very important. Hence, the purpose of this paper was purification of an extract obtained from Dendrobium officinale Kimura et Migo and cadmium removal using thermoplastic starch (S1), modified TPS with poly (butylene succinate); 25% of TPS + 75% PBS (S2); 50% of TPS + 50% PLA (S3); and 50% of TPS + 50% PLA with 5% of hemp fibers (S4), as well as ion exchangers of different types, e.g., Lewatit SP112, Purolite S940, Amberlite IRC747, Amberlite IRC748, Amberlite IRC718, Lewatit TP207, Lewatit TP208, and Purolite S930. This extract is used in cancer treatment in traditional Chinese medicine (TCM). Attenuated total reflectance-Fourier transform infrared spectroscopy, thermogravimetric analysis with differential scanning calorimetry, X-ray powder diffraction, gel permeation chromatography, surface analysis, scanning electron microscopy with energy dispersive X-ray spectroscopy, and point of zero charge analysis were used for sorbent and adsorption process characterization, as well as for explanation of the Cd(II) sorption mechanism.

RNA sequencing-based exploration of the effects of far-red light on lncRNAs involved in the shade-avoidance response of D. officinale.

Dendrobium officinale (D. officinale) is a valuable medicinal plant with a low natural survival rate, and its shade-avoidance response to far-red light is as an important strategy used by the plant to improve its production efficiency. However, the lncRNAs that play roles in the shade-avoidance response of D. officinale have not yet been investigated. This study found that an appropriate proportion of far-red light can have several effects, including increasing the leaf area and accelerating stem elongation, in D. officinale. The effects of different far-red light treatments on D. officinale were analysed by RNA sequencing technology, and a total of 69 and 78 lncRNAs were differentially expressed in experimental group 1 (FR1) versus the control group (CK) (FR1-CK) and in experimental group 4 (FR4) versus the CK (FR4-CK), respectively. According to GO and KEGG analyses, most of the differentially expressed lncRNA targets are involved in the membrane, some metabolic pathways, hormone signal transduction, and O-methyltransferase activity, among other functions. Physiological and biochemical analyses showed that far-red light promoted the accumulation of flavonoids, alkaloids, carotenoids and polysaccharides in D. officinale. The effect of far-red light on D. officinalemight be closely related to the cell membrane and Ca2+ transduction. Based on a Cytoscape analysis and previous research, this study also found that MSTRG.38867.1, MSTRG.69319.1, and MSTRG.66273.1, among other components, might participate in the far-red light signalling network through their targets and thus regulate the shade-avoidance response of D. officinale. These findings will provide new insights into the shade-avoidance response of D. officinale.

Differences in the chemical composition of Dendrobium officinale Kimura et Migo and Dendrobium crepidatum Lindl based on UPLC-Q-TOF-MS/MS and metabolomics / 药学学报

italic>Dendrobium officinale Kimura et Migo is a rare Chinese herbal medicine, while Dendrobium crepidatum Lindl is a local medicine in Yunnan, both of which have the function of nourishing yin and stomach. To reveal the differences in chemical composition between the two species, ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was used to analyze the chemical composition of stems and leaves of D. officinale and D. crepidatum. Principal component analysis (PCA) and partial least squares discriminant analysis (OPLS-DA) were used to determine the differences in metabolites between species and parts of Dendrobium. Fifty-eight chemical compounds were identified in the two species. Analysis indicated that the side ring of alkaloids connected with nitrogen was readily cleaved during analysis. The results of PCA analysis showed that the stems and leaves of D. officinale and D. crepidatum could be easily differentiated, and the chemical constituents of D. officinale and D. crepidatum were significantly different. OPLS-DA analysis showed that there were 16 metabolite differences between the stems and 22 differences in metabolites between the leaves of D. officinale and D. crepidatum. The main metabolite differences in components between the two Dendrobium species were dendrocrepidine B, dendrocrepidine C and dendrocrepine. There were 14 differences in metabolites between the stems and leaves of D. crepidatum. In conclusion, the chemical compositions of D. officinale and D. crepidatum are quite different; the small molecular compounds of D. officinale are mainly terpenoids and flavonoids, and the content of alkaloids is low. There is no significant difference between stem and leaf. In contrast, D. crepidatum is mainly composed of alkaloids and terpenoids, with crepidamine and dendrocrepine as its unique components, and there are great differences in the components between stems and leaves. This study provides a theoretical basis for the development and utilization of Dendrobium resources.

Advances and prospects of pharmacological activities of Dendrobium officinale Kimura et Migo polysaccharides / 药学学报

italic>Dendrobium officinale Kimura et Migo (D. officinale) has been used as a valuable traditional Chinese medicine for more than 2 000 years in China. Modern research has confirmed a wide range of pharmacological activities, such as regulating blood sugar, improving gastrointestinal inflammation, and regulating immunity. Polysaccharides are the main active ingredients of D. officinale. With the intensive studies of the pharmacological activities of D. officinale, evidence for the pharmacological effects and potential mechanisms of D. officinale polysaccharides has increased dramatically. In this review, we summarized the latest progress in the pharmacological and mechanical studies of D. officinale polysaccharides, and based on the pharmacological efficacy and oral absorption and utilization characteristics of D. officinale polysaccharides, it is proposed that regulating the gut microbiota may be one of the key mechanisms for D. officinale to exert its beneficial effects. Research on the mechanism of D. officinale polysaccharides puts forward new research directions and prospects.

Research progress on genomics, transcriptomics and functional genes of Dendrobium officinale / 中草药

Dendrobium officinale is one of the most precious medicinal plants in the family of Orchidaceae, and rich in polysaccharides, astragalus, bibenzyls and alkaloids. It has effects such as antioxidant, anti-cancer, immuno-enhancing, lowering blood sugar, and alleviating liver injury. Its huge medicinal, scientific and commercial value has raised a research upsurge, especially in the past five years, nucleic acid molecular biology of Dendrobium officinale has made more and more exciting results. In order to provide scientific guidance for further research on functional genes, this paper reviews the recent progress in research on genomics, transcriptomics and functional genes of D. officinale.

Cloning and expression analysis of a novel alkaline/neutral invertase (DoNI2) gene in Dendrobium officinale / 中草药

Objective: To clone novel member of alkaline/neutral invertase (NI) gene in a rare and endangered medicinal plant of Dendrobium officinale, conduct bioinformatic analysis and detect the quantitative expression in different organs. Methods: Primers were designed according to NI gene segment which was selected from leaf transcriptome sequencing results of D. officinale. The full-length cDNA of NI gene was cloned via homology-based cloning and rapid amplification of cDNA ends (RACE) approach. The physical and chemical properties, secondary structure and tertiary structure of NI protein were forecasted and analyzed using related software. The expression levels of NI gene in roots, stems, and leaves of D. officinale were detected using real-time PCR. Results: A novel gene encoding a NI protein was cloned from D. officinale. This gene (named as DoNI2, GenBank accession number: KY794404) had a total length of 2 397 bp with an open reading frame of 1 836 bp, and encoded a predicted polypeptide of 611 amino acids with a molecular weight of 69 050. Bioinformatics predicted that the isoelectric point of DoNI2 gene encoding protein was 6.38, the instability coefficient was 44.95, and the hydrophobic coefficient was −0.232. RT-PCR showed that DoNI2 gene expressed in all organs with highest expression level in stems and the lowest in roots. DoNI2 gene expression was significantly positively correlation with NI enzymatic activities at different growth years of D. officinale. Conclusion: The full length cDNA sequence in a mitochondrial DoNI2 gene was identified, facilitating future functional analysis of the gene involving in the regulation of sugar metabolism in D. officinale.

Cloning and expression analysis of gene DoSRK2E in Dendrobium officinale / 中草药

Objective To clone the SnRK2 gene in Dendrobium officinale and investigate its characteristics and expression pattern. Methods RT-PCR and RACE techniques were used to clone the full-length cDNA of DoSRK2E, with the aids of a series of online bioinformatic software, characteristics including molecular mass, isoelectric point, conserved domain, transmembrane structure, signal peptide, and subcellular localization of the deduced protein were analyzed. Besides, the sequence of the deduced protein was aligned with those of other plant SnRK2 by DNASTAR, and phylogenetic relationships were reconstructed utilizing MEGA. Finally, tissue specific expression pattern of DoSRK2E was tested by quantitative real-time PCR (qRT-PCR). Results The full-length cDNA of DoSRK2E (GenBank accession API65110) is 1 795 bp with a 1 086 bp complete open reading frame (ORF). The predicted molecular mass and isoelectric point of the deduced protein DoSRK2E were 40 850 and 4.80, respectively. No signal peptide nor transmembrane region were detected, this protein contains one protein kinase domain, one ATP binding site, and one Ser/Thr active site, which was predicted most likely to be located in the endoplasmic reticulum membrane. DoSRK2E protein showed high similarity with those from other plant SnRK2, and its phylogenetic location was in Group III of SnRK2 subfamily, and phylogenetically closest to AtSnRK2.6 from Arabidopsis. In addition, qPCR analysis revealed that DoSRK2E showed the highest expression level in root, followed by stem and leaf. Conclusion A SnRK2 family gene DoSRK2E was cloned from the rare and endangered medicinal plant D. officinale for the first time. The Characteristics and expression pattern of this gene were analyzed. This study will provide a basis for further exploration of the regulation mechanisms of DoSRK2E in D. officinale under stress conditions.

Distinguishing the Chinese materia medica Tiepishihu from similar Dendrobium species of the same genus using histological and microscopic method.

The Chinese Materia Medica, Tiepishihu, used as a tonic for over one thousand years, is a well-known precious medicine in China. According to the Chinese Pharmacopoeia, its source is the species Dendrobium officinale Kimura et Migo, which is distinguished from other species in Dendrobium genus. However, these species from the same genus are similar with Tiepishihu and caused confusion in the market. To find a quick and simple method to distinguish Tiepishihu from other similar species, histologic and microscopic methods were combined together to investigate the transverse section of stem of Tiepishihu and other similar species. Phloroglucinol test solution with hydrochloric acid was used to reveal the lignified tissue by staining the transverse section of Tiepishihu and similar species. Results revealed the unique identification characteristics to distinguish Tiepishihu from similar species, which were difficult to distinguish by other methods. The identification characteristics of Tiepishihu include the cells of vascular bundle sheath were stained red, parenchyma cells were not stained red. What's more, other species can be distinguished from each other with microscopic and histological characteristics. These characteristics proved stable and can be easily observed by normal light microscopic examination. This method is rapid, accurate, stable, and inexpensive.

Cloning and preliminary expression analysis on HDS gene in Dendrobium officinale / 中草药

Objective: To clone the 4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase (HDS) gene from Dendrobium officinale, induce fusion protein in Escherichia coli, and explore the regular pattern about HDS gene in different tissues of D. officinale. Methods: RT-PCR and RACE technologies were used to clone the full-length cDNA of DoHDS. Using relevant softwares and online sites to analyze bioinformatics. Then the expression patterns of DoHDS were studied by real-time PCR. Constructing prokaryotic expression vector pET-28a (+)-DoHDS to induce the expression protein in E. coli BL21 (DE3). Results: The DoHDS gene was successfully obtained (GenBank accession number KJ161312), the full-length cDNA was 2666 bp and ORF was 2238 bp, coding the protein containing 745 amino acids. Relative real-time PCR analysis indicated that DoHDS showed the higher transcript abundance in the stems, 5 fold higher than protocorm-like bodies. The SDS-PAGE results showed that a relative molecular weight of 82700 recombinant protein was produced. Conclusion: The cDNA encoding HDS from D. officinale is cloned. The prokaryotic expression vector and different tissues expression patterns of DoHDS are constructed. It is helpful for the future research on the mechanism of terpenoid biosynthesis in medicinal plants D. officinale.

Cloning and expression analysis of gibberellin 3-oxidase gene in Dendrobium officinale / 中草药

Objective: To clone the gibberellin 3-oxidase gene DoGA3ox from an important and endangered medicinal plant Dendrobium officinale (Orchidaceae), followed by bioinformatic and expression analysis. Methods: RACE and RT-PCR were used to isolate the full-length gene. The physicochemical properties, conserved domains, and subcellular localization of DoGA3ox protein were determined using a series of bioinformatic tools. The phylogenetic analyses were performed using DNASTAR 7.0 and MEGA 6.0 software. Real time quantitative PCR was employed for gene expression analysis. Results: The full-length cDNA of DoGA3ox (GenBank registration number KT597694) was 1 318 bp in size, and encoded a 353-amino acid peptide chain with a molecular weight of 39052.5 and an isoelectric point (pI) of 6.21; The deduced DoGA3ox protein without transmembrane or signal peptide residues, contained the gibberellin 3-oxidases conserved domains that DIOX_N (40-130) and 2OG-FeII_Oxy (197-299). DoGA3ox had 51%-56% similarity with GA3ox proteins from various plants, and was closely related to the monocot Allium fistulosum, Elaeis guineensis, and Phoenix dactylifera. The relative transcript levels of DoGA3ox were increased at stage 1, then decreased (stage 2), and increased again (stage 3) during D. officinale symbiotic/asymbiotic seed germination, the fold change to ungerminated seeds was 13.44 (symbiotic)/5.21 (asymbiotic), 7.28/2.32 and 9.40/6.21, respectively. Moreover, its transcript level was higher in symbiotic germination than that in the asymbiotic status. Conclusion: The full-length DoGA3ox gene is cloned for the first time, and its expression in different stages during seed germination indicates that DoGA3ox gene plays a crucial role in the regulation of seed germination in D. officinale.

Cloning and expression analysis of pyruvate kinase gene in Dendrobium officinale / 中草药

Objective: To clone the pyruvate kinase (PK) gene DoPK in a rare and endangered medicinal plant Dendrobium officinale, followed by bioinformatic analysis and to detect the expression in different organs. Methods: RT-PCR and RACE technologies were used to clone the full length cDNA of DoPK gene. The molecular characteristics such as physiochemical properties, conserved domain, and sub-cellular localization of the deduced DoPK protein were determined using a series of bioinformatic tools. The analyses of multiple alignment and phylogenetic tree were performed using DNASTAR 6.0 and MEGA 4.0 softwares, respectively. Real time quantitative PCR (RT-qPCR) was used for gene expression analysis in different organs. Results: The full length cDNA of DoPK was 1895 bp in length (GenBank accession KC178572) and encoded the protein of 511 amino acids with a molecular weight of 55040 and an isoelectric point (PI) of 7.00. Sequence analysis demonstrated that DoPK was similar to PK genes from Selaginella moellendorffii, Arabidopsis thaliana, Solanum tuberosum, and Vitis vinifera with the identities of 71%, 86%, 89%, and 91%, respectively. The deduced DoPK protein contained the conserved PK domain (21-497) and the active site (235-247). Phylogenetic analysis indicated that DoPK was grouped into the CYTOSOLIC-1 subfamily and was closely related to monocots. The transcription level of DoPK in the stems of D. officinale was the highest (2.29-fold higher than that in the leaves), followed by that in the roots (1.28-fold). Conclusion: The full length cDNA sequence in a CYOTOSLIC DoPK gene is identified, facilitating further functional analysis of the gene involving in the growth and development of D. officinale.

Analysis of the Dendrobium officinale transcriptome reveals putative alkaloid biosynthetic genes and genetic markers.

Dendrobium officinale Kimura et Migo (Orchidaceae) is a traditional Chinese medicinal plant. The stem contains an alkaloid that is the primary bioactive component. However, the details of alkaloid biosynthesis have not been effectively explored because of the limited number of expressed sequence tags (ESTs) available in GenBank. In this study, we analyzed RNA isolated from the stem of D. officinale using a single half-run on the Roche 454 GS FLX Titanium platform to generate 553,084 ESTs with an average length of 417 bases. The ESTs were assembled into 36,407 unique putative transcripts. A total of 69.97% of the unique sequences were annotated, and a detailed view of alkaloid biosynthesis was obtained. Functional assignment based on Kyoto Encyclopedia of Genes and Genomes (KEGG) terms revealed 69 unique sequences representing 25 genes involved in alkaloid backbone biosynthesis. A series of qRT-PCR experiments confirmed that the expression levels of 5 key enzyme-encoding genes involved in alkaloid biosynthesis are greater in the leaves of D. officinale than in the stems. Cytochrome P450s, aminotransferases, methyltransferases, multidrug resistance protein (MDR) transporters and transcription factors were screened for possible involvement in alkaloid biosynthesis. Furthermore, a total of 1061 simple sequence repeat motifs (SSR) were detected from 36,407 unigenes. Dinucleotide repeats were the most abundant repeat type. Of these, 179 genes were associated with a metabolic pathway in KEGG. This study is the first to produce a large volume of transcriptome data from D. officinale. It extends the foundation to facilitate gene discovery in D. officinale and provides an important resource for the molecular genetic and functional genomic studies in this species.

Effect of Dendrobium officinale Kimura et Migo on substance metabolism and exercise capacity in rats after exercise training / 中国药学杂志

OBJECTIVE: To study the effects of Dendrobium officinale Kimura et Migo on the rats substance metabolism and anti-exercise fatigue capacity in the exercise training. METHODS: High-intensity endurance training rat was as a model. Divided the 6-week-old male (42 d age) Wistar rats 55 into 5 groups of 10 (removed the rats, which are not meeting the requirement): still group (group C), exercise group (group T), exercise ig low-dose Dendrobium officinale Kimura et Migo group of (group TML), exercise ig middle-dose of Dendrobium officinale Kimura et Migo group (group TMM), exercise ig high-dose of Dendrobium officinale Kimura et Migo group (group TMH). Gavage using professional device ig once a day, group TML, Group TMM, group TMH ig dose was 1.5, 3, 4. 5 g · kg-1 · d-1, the ig volume 5 mL · kg-1, Group C and group T were added to ig the same amount of saline. For 24 h after the last training, measured weight, exhaustive swimming time, Hemoglobin and other biochemical markers. RESULTS: The TM each weight no significant differences, and greater than the group T(P < 0.05) less than group C(P < 0.05). TM swimming time is longer than the group C and group T(P < 0.01), and the dose increased and extended. Prolonged exhaustive exercise lead to liver glycogen, muscle glycogen reserves decreased(group T(P < 0.01), groups TM(P < 0.05) lower than in group C. TM each group of liver glycogen [P < 0.05), inositol the original (P < 0.01] higher than the group T, no significant differences between each group of the TM, with the dose increase and higher). In addition, blood urea nitrogen rise[group T(P < 0.01) and groups TM(P < 0.05) blood urea nitrogen higher than in group C. Groups TM is lower than the group T(P < 0.05), TM each non-significant difference]. Hemoglobin decreased [group T(P < 0.01), groups TM(P < 0.05) lower than in group C. Groups TM is higher than the group T(P < 0.05), no significant difference between the TM each]. CONCLUSION: Dendrobium officinale Kimura et Migo supplement can promote protein synthesis, inhibition of amino acid and protein degradation, thereby increasing the hemoglobin content and glycogen reserves, enhanced resistance to fatigue, it has a multi-target, multi-channel features.