This study aimed to explore the mechanism of Cistanches Herba in the treatment of cancer-induced fatigue(CRF) by network pharmacology combined with in vivo and in vitro experiments to provide a theoretical basis for the clinical medication. The chemical constituents and targets of Cistanches Herba were searched from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). The targets of CRF were screened out by GeneCards and NCBI. The common targets of traditional Chinese medicine and disease were selected to construct a protein-protein interaction(PPI) network, followed by Gene Ontology(GO) functional and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses. A visual signal pathway rela-ted to Chinese medicine and disease targets was constructed. The CRF model was induced by paclitaxel(PTX) in mice. Mice were divided into a control group, a PTX model group, and low-and high-dose Cistanches Herba extract groups(250 and 500 mg·kg~(-1)). The anti-CRF effect in mice was evaluated by open field test, tail suspension test, and exhaustive swimming time, and the pathological morphology of skeletal muscle was evaluated by hematoxylin-eosin(HE) staining. The cancer cachexia model in C2C12 muscle cells was induced by C26 co-culture, and the cells were divided into a control group, a conditioned medium model group, and low-, medium-, and high-dose Cistanches Herba extract groups(62.5, 125, and 250 μg·mL~(-1)). The reactive oxygen species(ROS) content in each group was detected by flow cytometry, and the intracellular mitochondrial status was evaluated by transmission electron microscopy. The protein expression levels of hypoxia-inducible factor-1α(HIF-1α), BNIP3L, and Beclin-1 were detected by Western blot. Six effective constituents were screened out from Cistanches Herba. The core genes of Cistanches Herba in treating CRF were AKT1, IL-6, VEGFA, CASP3, JUN, EGFR, MYC, EGF, MAPK1, PTGS2, MMP9, IL-1B, FOS, and IL10, and the pathways related to CRF were AGE-RAGE and HIF-1α. Through GO enrichment analysis, it was found that the main biological functions involved were lipid peroxidation, nutrient deficiency, chemical stress, oxidative stress, oxygen content, and other biological processes. The results of the in vivo experiment showed that Cistanches Herba extract could significantly improve skeletal muscle atrophy in mice to relieve CRF. The in vitro experiment showed that Cistanches Herba extract could significantly reduce the content of intracellular ROS, the percentage of mitochondrial fragmentation, and the protein expression of Beclin-1 and increase the number of autophagosomes and the protein expression of HIF-1α and BNIP3L. Cistanches Herba showed a good anti-CRF effect, and its mechanism may be related to the key target proteins in the HIF-1α signaling pathway.
Animals ; Mice ; Cistanche ; Network Pharmacology ; Beclin-1 ; Reactive Oxygen Species ; Plant Extracts ; Drugs, Chinese Herbal/pharmacology* ; Molecular Docking Simulation ; Medicine, Chinese Traditional ; Neoplasms/genetics*

This study aims to establish a method for analyzing the chemical constituents in Cistanches Herba by high performance liquid chromatography(HPLC) and quadrupole-time-of-flight tandem mass spectrometry(HPLC-Q-TOF-MS/MS), and to reveal the pharmacological mechanism based on network pharmacology for mining the quality markers(Q-markers) of Cistanches Herba. The chemical constituents of Cistanche deserticola and C. tubulosa were analyzed via HPLC-Q-TOF-MS/MS. The potential targets and pathways of Cistanches Herba were predicted via SwissTargetPrediction and DAVID. The compound-target-pathway-pharmacological action-efficacy network was constructed via Cytoscape. A total of 47 chemical constituents were identified, involving 95 targets and 56 signaling pathways. We preliminarily elucidated the pharmacological mechanisms of echinacoside, acteoside, isoacteoside, cistanoside F, 2'-acetylacteoside, cistanoside A, campneoside Ⅱ, salidroside, tubuloside B, 6-deoxycatalpol, 8-epi-loganic acid, ajugol, bartsioside, geniposidic acid, and pinoresinol 4-O-β-D-glucopyranoside, and predicted them to be the Q-markers of Cistanches Herba. This study identified the chemical constituents of Cistanches Herba, explained the pharmacological mechanism of the traditional efficacy of Cistanches Herba based on network pharmacology, and introduced the core concept of Q-markers to improve the quality evaluation of Cistanches Herba.
Chromatography, High Pressure Liquid/methods* ; Cistanche ; Drugs, Chinese Herbal/pharmacology* ; Network Pharmacology ; Tandem Mass Spectrometry/methods*

Cistanches Herba(CH), a valuable medicinal material which has long been used, originated from Shennong's Classic of Materia Medica. It has a wealth of names, such as Rousongrong, Heisiming, Dijing, and Dayun. The definition on the original plants which are parasitic and distributed in the unique environments in remote areas has been confusing, resulting in the emergence of various counterfeits and substitutes. Moreover, the records on the property, flavor, tropism, and indications of CH are also different. In order to further explore the cultural connotation and medicinal value of CH and further clarify its source and harvesting and processing methods, this study conducted further classical literature research on its name, harvesting and processing, property and flavor, meridian tropism, efficacy and clinical use, and textual research on its source and habitats, providing a reference for research, clinical medication, development and utilization, and industry development of CH.
Cistanche ; Materia Medica ; Publications ; Drugs, Chinese Herbal

The current study aims to rapidly and comprehensively profile the chemical composition of Cistanche salsa using direct infusion coupled with MS/MS~(ALL)(DI-MS/MS~(ALL)). The C. salsa extract was directly imported into electrospray ionization(ESI) source of quadrupole time-of-flight(Q-TOF) mass spectrometer with an infusion pump at a flow rate of 10 μL·min~(-1). Acquisition program was applied under negative ionization polarity to collect one MS~1 spectrum(m/z 50-1 200), followed by 1 150 MS~2 spectra with precursor isolation window(m/z 1) amongst mass range m/z 50-1 200. After each MS~2 spectrum was matched to its precursor ion, putative identification was conducted through matching mass spectral data with literature and database. A total of 31 components were identified from C. salsa, including 9 phenylethanoid glycosides, 2 iridoids, 4 saccharides, 9 organic acids, and 7 other compounds, similar to those from C. tubulosa and C. deserticola. In conclusion, DI-MS/MS~(ALL), a facile and reliable analytical tool, can be employed for qualitative analysis of chemical constituents in C. salsa. The research offers a promising strategy to achieve rapid chemome profiling of herbal medicine and provides an alternative source of Cistanches Herba.
Chromatography, High Pressure Liquid ; Cistanche ; Drugs, Chinese Herbal ; Glycosides ; Plants, Medicinal ; Tandem Mass Spectrometry

Several Orobanche medicinal plants sometimes served as alternative sources of Cistanches Herba, attributing to the benefits such as tonifying kidney, strengthening tendons and bones. Among them, O. coerulescens, O. cernua and O. pycnostachya have been widely utilized in northern China for treatments of pains in the loins and knees, impotence, and spermatorrhea. However, their chemical profiles haven't been elucidated. In the present study, UHPLC-IT-TOF-MS was implemented to conduct in-depth chemome profiling of O. coerulescens, O. cernua and O. pycnostachya, aiming to achieve a comprehensive chemical characterization and to provide pronounced information for the quality control and clinical applications. An ACE Ultra-Core 2.5 Super C_(18)(3.0 mm×150 mm, 2.5 μm) column was deployed for chromatographic separations, and high-resolution MS~n spectra were recorded by IT-TOF-MS. Forty-eight components, in total, were observed, and thirty-eight ones were structurally annotated according to proposing mass fragmentation patterns, matching with relevant databases. Particularly, nine ones were confirmed by reference compounds. Overall, the chemical compositions of O. coerulescens and O. cernua are quite similar, and differences occur between O. pycnostachya and the prior two ones; primary chemical family is phenylethanoid glycosides, and several lignan glycosides as well as iridoid glycosides are also observed; the primary components include acteoside, isoacteoside, crenatoside and 2'-acetylacteoside, etc.
China ; Cistanche ; Glycosides ; Male ; Orobanche ; Plants, Medicinal

To investigate the inhibitory effects and mechanism of Cistanche tubulosa ethanol extract( CTEE) against oxygen-glucose deprivation/reperfusion( OGD/R)-induced PC12 cells neuronal injury. In this study,OGD/R-induced PC12 cells were used to explore the neuroprotective effects of CTEE( 12. 5,25,50 mg·L-1) by detecting cell viability with MTT assay,apoptosis with AO/EB and Hoechst 33258,mitochondrial membrane potential changes with JC-1 staining,mitochondrial oxidative stress with MitoSOX staining,as well as the apoptosis-related protein expression( PARP,cleaved PARP,caspase-3,cleaved caspase-3,Bax,Bcl-2) with Western blot. RESULTS:: showed that CTEE effectively protected OGD/R-induced neuronal injury and increased the survival rate of PC12 cells.AO/EB and Hoechst 33258 staining showed that CTEE could effectively inhibit apoptosis. Moreover,JC-1 and MitoSOX staining results showed that CTEE decreased mitochondrial stress and mitochondrial membrane potential imbalance in PC12 cells in a concentration-dependent manner. Meanwhile,CTEE could obviously suppress the activation of key proteins in mitochondrial apoptosis pathway such as caspase-3 and PARP,and significantly inhibit the rise of Bax and down-regulation of Bcl-2. In conclusion,CTEE has obvious protective effects on OGD/R-induced PC12 cells neuronal injury,potentially via inhibiting mitochondrial oxidative stress and apoptosis-related signaling pathway.
Animals ; Apoptosis ; Caspase 3 ; metabolism ; Cistanche ; chemistry ; Ethanol ; Glucose ; Neuroprotective Agents ; pharmacology ; Oxidative Stress ; Oxygen ; PC12 Cells ; Plant Extracts ; pharmacology ; Poly (ADP-Ribose) Polymerase-1 ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; bcl-2-Associated X Protein ; metabolism

Based on UPLC specific chromatogram and determination of seven main components,this study aimed at evaluating the quality of Cistanche deserticola,C. tubulosa and C. sinensis. Echinacoside,cistanoside A,verbascoside,tubuloside A,isoacteoside,2'-acetylacteoside,tubuloside B were used as reference substances. UPLC analysis was performed on a Waters ACQUITY UPLC HSS T3 column( 2. 1 mm×100 mm,1. 8 μm). The mobile phase was acetonitrile-0. 08% trifluoroacetic acid solution. The flow rate was0. 3 mL·min-1,and the injection amount was 10 μL. The column temperature was 40 ℃,and the detection wavelength was 330 nm.The UPLC specific chromatograms were processed with ChemPattern software. UPLC specific chromatograms of C. deserticola and C.tubulosa from different samples were of high similarity,but the similarities of their counterfeit C. sinensis were less than 0. 06. Both of cluster and principal component analysis can distinguish certified products and counterfeits. The content ratios of echinacoside/verbascoside and verbascoside/isoacteoside were quite different between C. deserticola and C. tubulosa,which had distinct significance.The UPLC specific chromatogram and contents of seven main components can provide a basis for quality evaluation of Cistanches Herba.
Chromatography, High Pressure Liquid ; Cistanche ; chemistry ; classification ; Drugs, Chinese Herbal ; analysis ; standards ; Phytochemicals ; analysis ; Principal Component Analysis

In this study, taking Cistanche deserticola in Xinjiang as the experimental material, the optimal process for extracting polysaccharides from C. deserticola with water extraction was studied by using single factor and orthogonal experiment. Its effects on protein removal and polysaccharides retaining were investigated by using Sevag, enzymatic method or combination of these two methods, so as to determine the optimal method for protein removal from polysaccharides of C. deserticola; the decolorization and purification methods such as macroporous resin of AB-8 and activated Carbon were used to determine the optimal process. The results showed that the extraction rate of polysaccharides from C. deserticola was 18.40% during the optimal process of the water extraction as follows: extraction temperature 75 ℃, extraction time 165 min and solid-liquid ratio 1∶55. The protein removal rate can reach 31.40% and polysaccharide retention rate can reach 96.00% under the optimal protein removal process: temperature 50 ℃, time 2 h, and papain dosage 0.2%. The decolorization rate of activated Carbon and macroporous resin called AB-8 was 80.37% and 86.43%, and the recovery rate of polysaccharides was 77.05% and 91.93%, respectively, suggesting that macroporous resin was more suitable for decoloration. Macroporous resin named AB-8 increased the purity of the polysaccharide crude extract from 67.70% to 84.80% under the following conditions: concentration of the sample 4 g·L~(-1), concentration of the eluent 60% ethanol, and the flow rate 1 mL·min~(-1), showing significant purification effect.
Cistanche ; chemistry ; Plant Extracts ; chemistry ; Polysaccharides ; isolation & purification ; Temperature ; Water

To intuitively understand the phenotypic diversity of intra-population and inter-population of the medicinal Cistanche Herba distributed in Xinjiang province, three species of Cistanche Herba were selected for the first time to be conducted to phenotypic observation and measurement from the morphological perspective, aiming to fill the gap in the morphological research concerning Cistanche Herba, and discuss about the relationship between the phenotypic variation and the host plants together with the geographical conditions, thus better understanding the speciation and evolutionary mechanism of Cistanche Herba and providing some scientific basis for the resource protection and germplasm breeding of Cistanche Herba. Based on sampling survey, a total of 118 well grown medicinal Cistanche samples from 17 Cistanche Herba distribution areas in Xinjiang province were selected, and various indexes were observed and measured. Besides, ANOVA and clustering analysis were conducted with 9 phenotypic quantity characters. The Cistanche Herba was plentiful in phenotypic variation. For detail, significant intra-population differences were observed in eight of the nine character indexes, and the intra-population differences were more obvious than those of inter-population. For each quantity character of the three species, the flower density possessed the maximal variable coefficient (71.1%) while the flower length was the minimum (15.9%). The phenotypic variation was also obvious among different populations. Specifically, the average variable coefficient of flower number was the maximal one (46.5%) and the flower length was the minimum one (10.0%). For different populations, the average variable coefficient of the D4 population was the maximal one (41.1%) and the S3 population was the minimum one (13.5%). According to the clustering analysis, all the samples of Cistanche Herba could be divided into three groups when the Euclidean distance was set at 15. The populations of S1, S3, D1, S2, D4, D6, D7 and D5 which distributed in the north of Xinjiang were clustered into one group, and the populations of D8, S4, D9, D2 and D3 that grown in east and central of Xinjiang were clustered into another group. The populations of C. deserticola and C. salsa could not be completely separated, but both of them were obviously differentiated from the T1, T3 and T2 populations of C. tubulosa. Besides, the C. deserticola and C. salsa displayed a patch distribution among different populations, and similar phenotypic characters were shared for each population. The research results of phenotype were consistent with that of molecular biology study of Cistanche Herba. The different phenotypic characters in different distribution areas were deduced to be arose from geographical isolation caused by mountains, which led to the specific genetic structure for each population of Cistanche Herba during the long-term adaptation and evolution. In conclusion, the current study showed the adaptation potency of Cistanche Herba exposed to different habitats.
China ; Cistanche ; classification ; Cluster Analysis ; Ecosystem ; Flowers ; Genetics, Population ; Phenotype ; Plants, Medicinal ; classification

The genus Orobanche, Cistanche and Boschniakia are taxonomically classified as members of Orobanchaceae. The medicinal plants of these three genera are closely related, and their traditional curative effects are similar. As representative compounds, phenethyl glycosides are predominantly dominant type both in amount and in chemical structural varieties, which are considered to be the important bioactive material basis of these genera. In this paper, phenethyl glycosides and their pharmacological activities are described in a single list. In addition, the other compounds were also reviewed in order to better compare the difference of the bioactive substances. These findings have important reference value for effective development and rational utilization of resources of medicinal plants in the family Orobanchaceae.
Cistanche ; Glycosides ; Orobanchaceae ; Orobanche ; Plants, Medicinal