Study of the therapeutic effect of raw and processed Vladimiriae Radix on ulcerative colitis based on intestinal flora,metabolomics and tissue distribution analysis

BackgroundThe intestinal flora imbalance and metabolic disorders are closely related to the pathogenesis of ulcerative colitis (UC). As a commonly used herb for the treatment of gastrointestinal diseases, Vladimiriae Radix (VR) has been used for hundreds of years, and its main active ingredients are costunolide (COS) and dehydrocostus lactone (DEH). Clinical usage habits and previous studies have shown that the processed Vladimiriae Radix (pVR) seems to be more suitable for treating bowel disease than the raw Vladimiriae Radix (rVR), but there is still no relevant comparative study.PurposeTo investigate the therapeutic effect of rVR and pVR on UC by analyzing the intestinal flora, metabolomics and tissue distribution.MethodsUC rat models were established to investigate the anti-inflammatory activities of rVR and pVR by enzyme-linked immunosorbent assay (ELISA), and to study their regulation of intestinal flora and metabolism by 16s rRNA gene analysis and Ultra Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS). Moreover, the distribution of COS and DEH in UC mouse tissues were also observed by High Performance Liquid Chromatography Mass Spectrometry (HPLC-MS).ResultsrVR and pVR reduced tissue damage and the levels of TNF-α, IL-6, IL-1β, IL-10, TGF-β and MPO, especially pVR. 16s rRNA gene analysis showed that rVR superior in ameliorating species evenness and restoring the abundance of Lachnospiraceae and Ruminococcaceae, while pVR is better at increasing the richness and the abundance of Prevotellaceae. Metabolomics analysis suggested that rVR regulates the β-alanine, pantothenic acid and coenzyme A biosynthesis, but pVR regulates more abundant metabolic pathways. The tissue distribution data indicated the accumulation of COS and DEH in the gastrointestinal tract.ConclusionrVR and pVR had obvious therapeutic effect on UC. The potential mechanisms might be regulating abnormal metabolism, affecting the diversity and structure of intestinal flora, and accumulation of COS and DEH in colon.     

Two carabrane-type sesquiterpenes from Vladimiria souliei

Two new carabrane sesquiterpenes, 4,8-dioxo-6β-methoxy-7β,11-epoxy carabrane (1), and its isomer, 4,8-dioxo-6β-methoxy-7α,11-epoxy carabrane (2) were isolated from the roots of Vladimiria souliei. Their structures were elucidated by spectroscopic methods (IR, ESI-MS, HR-ESI-MS, 1D and 2D NMR).     

Dolomiaea souliei ethyl acetate extract protected against α-naphthylisothiocyanate-induced acute intrahepatic cholestasis through regulation of farnesoid x receptor-mediated bile acid metabolism

BackgroundCholestasis is characterized by accumulation of bile components in liver and systemic circulation. Restoration of bile acid homeostasis via activating farnesoid x receptor (FXR) is a promising strategy for the treatment of cholestasis. FXR-SHP (small heterodimer partner) axis plays an important role in maintaining bile acid homeostasis.PurposeTo investigate the anti-cholestasis effect of Dolomiaea souliei (Franch.) C.Shih (D. souliei) and clarify its underlying mechanism against α-naphthylisothiocyanate (ANIT) induced acute intrahepatic cholestasis.MethodsANIT-induced Sprague-Dawley rats were employed to investigate the anti-cholestasis effect of D. souliei ethyl acetate extract (DSE). Ursodeoxycholic acid (UDCA) was used as positive control. Bile flow and blood biochemical parameters were measured. Liver histopathological examination was conducted via hematoxylin-eosin staining. Western blot analysis was carried out to evaluate the protein levels related to bile acids metabolism and inflammation. The interactions between FXR and costunolide or dehydrocostus lactone, were conducted by molecular docking experiments. The effect of costunolide and dehydrocostus lactone on aspartate aminotransferase (AST), alanine aminotransferase (ALT) levels and FXR expression were also evaluated using guggulsterone-induced L02 cells.ResultsDSE could promote bile excretions and protect against ANIT-induced liver damage in cholestasis rats. Protein levels of FXR, SHP, Na⁺/taurocholate cotransporter (NTCP), bile salt export pump (BSEP), multidrug resistance-associated protein 2 (MRP2) were increased and the expressions of cholesterol 7α-hydroxylase (CYP7A1) and sterol 27-hydroxylase (CYP27A1) were decreased by DSE. Meanwhile, the anti-inflammatory factors, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) were also significantly increased, and the pro-inflammatory factor, interleukin-10 (IL-10), was significantly decreased in rats of DSE groups. Molecular docking revealed that costunolide and dehydrocostus lactone could be well docked into the FXR protein molecule, and hydrophobic interactions played the main function. Costunolide could reverse the increased AST and ALT levels and increase the FXR expression in guggulsterone-induced L02 cells.ConclusionDSE had an anti-cholestasis effect by activating FXR-SHP axis, inhibiting synthesis of bile acid, and increasing bile secretion, together with inflammatory response and improving liver injury. Costunolide may be the main active component. This study provided a potential therapeutic mechanism for D. souliei as an anti-cholestasis medicine in the treatment of cholestasis liver diseases.     

Use of a fluorescent imaging plate reader--based calcium assay to assess pharmacological differences between the human and rat vanilloid receptor

The cloned vanilloid receptor 1 (VR1) is a ligand-gated calcium channel that is believed to be the capsaicin-activated vanilloid receptor found in native tissues, based on similarities regarding molecular mass, tissue distribution, and electrophysiological properties. Using a Fluorescent Imaging Plate Reader (FLIPR), along with Fluo-3 to signal intracellular calcium levels ([Ca(++)](i)), rat VR1 (rVR1) and a human orthologue (hVR1) were pharmacologically characterized with various VR1 ligands. HEK-293 cells, stably expressing rVR1 or hVR1, exhibited dose-dependent increases in [Ca(++)](i) when challenged with capsaicin (EC(50)s congruent with 10 nM). Responses to capsaicin were blocked by the VR1 antagonist capsazepine and were dependent on VR1 expression. Potencies for 10 structurally diverse VR1 agonists revealed rVR1 potencies highly correlated to that of hVR1 (R(2) = 0.973). However, a subset of agonists (tinyatoxin, gingerol, and zingerone) was approximately 10-fold more potent for rVR1 compared to hVR1. Schild analysis for blockade of capsaicin-induced responses by capsazepine was consistent with competitive antagonism, whereas ruthenium red displayed noncompetitive antagonism. Compared to rVR1, hVR1 was more sensitive to blockade by both antagonists. For both rVR1 and hVR1, time-response waveforms elicited by resiniferatoxin increased more gradually compared to other agonists. Tinyatoxin also displayed slow responses with hVR1 but showed rapid responses with rVR1. Thus, FLIPR technology can be used to readily reveal differences between rVR1 and hVR1 pharmacology with respect to potencies, efficacies, and kinetics for several VR1 ligands.     

Interaction between protein kinase Cmu and the vanilloid receptor type 1

The capsaicin receptor VR1 is a polymodal nociceptor activated by multiple stimuli. It has been reported that protein kinase C plays a role in the sensitization of VR1. Protein kinase D/PKCmu is a member of the protein kinase D serine/threonine kinase family that exhibits structural, enzymological, and regulatory features distinct from those of the PKCs, with which they are related. As part of our effort to optimize conditions for evaluating VR1 pharmacology, we found that treatment of Chinese hamster ovary (CHO) cells heterologously expressing rat VR1 (CHO/rVR1) with butyrate enhanced rVR1 expression and activity. The expression of PKCmu and PKCbeta1, but not of other PKC isoforms, was also enhanced by butyrate treatment, suggesting the possibility that these two isoforms might contribute to the enhanced activity of rVR1. In support of this hypothesis, we found the following. 1) Overexpression of PKCmu enhanced the response of rVR1 to capsaicin and low pH, and expression of a dominant negative variant of PKCmu reduced the response of rVR1. 2) Reduction of endogenous PKCmu using antisense oligonucleotides decreased the response of exogenous rVR1 expressed in CHO cells as well as of endogenous rVR1 in dorsal root ganglion neurons. 3) PKCmu localized to the plasma membrane when overexpressed in CHO/rVR1 cells. 4) PKCmu directly bound to rVR1 expressed in CHO cells as well as to endogenous rVR1 in dorsal root ganglia or to an N-terminal fragment of rVR1, indicating a direct interaction between PKCmu and rVR1. 5) PKCmu directly phosphorylated rVR1 or a longer N-terminal fragment (amino acids 1-118) of rVR1 but not a shorter one (amino acids 1-99). 6) Mutation of S116A in rVR1 blocked both the phosphorylation of rVR1 by PKCmu and the enhancement by PKCmu of the rVR1 response to capsaicin. We conclude that PKCmu functions as a direct modulator of rVR1.     

A preliminary study on pollination biology of Omphalogramma souliei Franch. (Primulaceae), a species endemic to China

The pollination biology of Omphalogramma souliei Franch., a species endemic to southwestern China, was investigated. Floral phenology, flower visitors, pollen/ovule ratio, attractants and rewards to the visitors were observed, measured, and recorded. Bagging experiments to exclude pollinators were carried out in the wild for two years. Our results revealed five important aspects of the reproductive biology of Omphalogramma souliei. 1) The pollen-ovule ratio was 1748±233. The breeding system was self-compatible, with facultative xenogamy. 2) The pollination syndrome is entomophily, and this species could not be pollinated by wind if the pollinators were unavailable. 3) Six insect species were observed visiting the flowers of Omphalogramma souliei in the wild, of them, three species of hymenoptera, Lasioglossum sp., Heriades parvula Cockerell and Micrapis florae Fabricius, are the principal floral visitors and effective pollinators. 4) The visual attractants to the visitors are floral color and shape, the large yellow anthers, and the rewards for visitors are pollen and nectar. 5) Cleistogamy may also occur, since the anthers of some flowers dehisced before opening of the corolla. The results of floral biology and pollination characters suggest that xenogamy predominated and autogamy played an assistant role in the evolution of reproduction and breeding system of O. souliei.     

Low genetic diversity and high genetic differentiation in the critically endangered Omphalogramma souliei (Primulaceae): implications for its conservation

Omphalogramma souliei Franch. is an endangered perennial herb only distributed in alpine areas of SW China. ISSR markers were applied to determine the genetic variation and genetic structure of 60 individuals of three populations of O. souliei in NW Yunnan, China. The genetic diversity at the species level is low with P=42.5% (percentage of polymorphic bands) and Hsp=0.1762 (total genetic diversity). However, a high level of genetic differentiation among populations was detected based on different measures (Nei's genetic diversity analysis: Gst=0.6038; AMOVA analysis: Fst=0.6797). Low level of genetic diversity within populations and significant genetic differentiation among populations might be due to the mixed mating system in which xenogamy predominated and autogamy played an assistant role in O. souliei. The genetic drift due to small population size and limited current gene flow also resulted in significant genetic differentiation. The assessment of genetic variation and differentiation of the endangered species provides important information for conservation on a genetic basis. Conservation strategies for this rare endemic species are proposed.     

Identification of a protein that interacts with the vanilloid receptor

The vanilloid receptor (VR1 or TRPV1) is a capsaicin (CAP)-sensitive non-selective cation channel. Although its channel activity is reportedly modulated through protein-protein interactions, to date very few VR1 interacting proteins have been identified. To address this issue, a yeast two-hybrid screening technique using the C-terminus of rVR1 as bait was employed. Upon interrogation of a mouse brain library, one gene product that interacts with VR1 and is highly homologous to human eferin was found. Its interaction with VR1 was confirmed by GST-pull-down and co-immunoprecipitation. When cotransfected into HEK cells, VR1 and eferin largely colocalize. Furthermore, in rat dorsal root ganglion cells, the rat eferin homologue also colocalizes with rVR1. However, this protein had no significant effect on VR1 channel activity in response to CAP. This was determined by two-electrode recording of oocytes and whole cell recording of HEK cells that were cotransfected with VR1 and human eferin.     

Therapeutic strategies of Moringa oleifera Lam. (Moringaceae) for stomach and forestomach ulceration induced by HCl/EtOH in rat model

BackgroundThe drumstick tree Moringa oleifera Lam. (Moringaceae), distributed in many parts of the world, is an important food plant with high nutritional value and used in medical applications and pharmaceutical industries. The aim of this study was to highlight the gastroprotective effect of Moringa oleifera in hydrochloric acid/Ethanol (HCl/EtOH) in a rat model.MethodsMoringa phytocompounds were characterized by infrared spectra (FTIR). Rats were induced for gastric ulcer with 150 mmol/L HCl/60% EtOH solution and pretreated orally with the edible infusion extract of the leaves of Moringa oleifera at a single dose of 100 mg/kg body weight (bw). Antioxidant parameters and lipid peroxide levels were measured and the pathological damage was histologically analysed.ResultsThe FTIR analysis showed the presence of several chemical biocompounds. The methanolic extract is the potent radical-scavengers with an estimated value of 87.54% at the higher concentration used (500 µg/ml) and antibacterial agent. Further, the DPPH inhibition value of the M. oleifera infusion was 80.58%. For in vivo analysis, mucus was highly produced in gastric mucosa of plant-treated rats, thereby pH were elevated in rats pretreated with M. oleifera compared to ulcerated animals. Whereas, lesion index was markedly reduced (79%) in stomach protected with plant. Interestingly, oral administration of M. oleifera protected gastric mucosa through decreasing MDA levels as well as increasing antioxidant enzyme activities (CAT, SOD, GPx).ConclusionOverall, the therapeutic value against acidified ethanol induced gastric and ulcer ability of M. oleifera might be due to its biocompounds.     

Genera Nova vel Minus Cognita Familiae Compositarum,I. Vladimiria Ilj., Diplazoptilon Ling et Dolomiaea DC.

The present paper concerns itself with three Composite genera of the tribe Cynareae (subtribe Carduinae), Vladimiria Ilj., Diplazoptilon Ling and Dolomiaea DC., all ranging throughout the Sino-himalayan region, These genera have in the past always been as sociated with the Eurasiatic genus Jurinea Cass., though there is hardly anything that can indicate such a relationship. Jurinea, according to the conception of Iljin, is a clearly deliminated natural genus, while the three genera now in question are all closely allied to Saussurea DC., being quite remote from Jurinea in the corolla forms, the structure of receptacle and pappus, and also in the morphology of pollen grains. Vladimiria was founded by Iljin, based on Jurinea salwinensis Hand.-Mzt. The genus is a close relative of Saussurea from which it differs chiefly in the usually many seriate pappus which are composed of scabrous or partly barbellulate bristles. As a re sult of our study, a tentative scheme of classification is here proposed, in which Iljin's original generic concept has been amplified to include a dozen of species previously re ferred to Jurinea by Franchet, Diels, Anthony and Handel-Mazzetti from the flora of south-western China and adjacent countries. Two sections, namely, sect. Sorocephalos Ling and sect. Vladimiria (Saussurea sect. Carduella Franch., Mazzettia Ilj.), are being proposed. Diplazoptilon, a new monotypic genus based upon Jurinea picridifolia Hand.-Mzt., is here proposed. It occupies an intermediate position between Vladimiria and Saussurea with a closer affinity to the former, from both of which it is, however, distinguished chiefly by the biseriate pappus with equal plumose bristles. Dolomiaea, a natural genus early founded by De Candolle on the basis of the Himalayan D. macrocephala DC., was considered by Bentham and Hooker f. as not different from Jurinea, a treatment having being followed since by most botanists of the past. But, as mentioned above, its relationship with Saussurea is obvious. It seems to differ from that genus in the two to many seriate pappus, consisting of numerous barbellulate bristles, and in the short style branches. In view of its affinity with Saussurea rather than with Jurinea, the generic status of Dolomiaea should be maintained accordingly. As construed above, the genus is now represented by about 5 species, most of which werepreviously referred to Jurinea by different botanists.     

Muscle loading and activation of the shoulder joint during humeral external rotation by pulley and variable resistance

BackgroundThe aim of the study was to evaluate differences in the loading of glenohumeral joint muscles between a cable pulley machine (CP) and variable resistance machine (VR) during axial humeral external rotation.MethodsEleven healthy male subjects took part in the study. Intramuscular electromyography from five muscles of the shoulder (medial deltoid, supraspinatus, infraspinatus and upper part of the trapezius), torque and power output was measured at different rotation angles and with different loads (10%, 50% and 100% of 1RM). Also the compressive and shear force in the glenohumeral joint was analyzed at the horizontal level at angles of rotation. External rotation was performed with a self-selected velocity on the scapular plane.FindingsIn the CP the range of movement became narrower than in the VR with increasing workload (P < 0.05). The activity of the infraspinatus did not grow in the CP after 50% load, while it did in the VR (P < 0.01). The upper part of the trapezius was activated less in the CP than in the VR (P < 0.01) machine when using 50% and 100% loads. In comparison with the CP, the shear forces that pull the head of the humerus in a posterior direction were more evenly distributed in the VR than in the CP at different angles of rotation (P < 0.001).InterpretationThe VR seems to make a broader range of motion possible, lager activation the primary external rotators and evenly distributed shear forces than the CP. However, performing the exercise with VR and high load also activates the upper part of the trapezius.RelevanceThese findings can be used in the development of exercise designs, methods and equipment for shoulder injury rehabilitation.     

川木香对实验性胃溃疡形成的抑制作用研究

目的:研究川木香对实验性胃溃疡形成的抑制作用.方法:采用利血平型小鼠胃溃疡模型、醋酸型大鼠胃溃疡模型,以雷尼替丁为对照药物,观察动物溃疡指数和溃疡抑制率.结果:川木香单体提取物(去氢木香内酯)、醋酸乙酯提取物、乙醇提取物,抑制利血平型溃疡存在统计学差异(与模型对照组比较,P＜0.01);醋酸乙酯提取物,抑制醋酸型溃疡存在统计学差异(与模型对照组比较,P＜0.05),其中高剂量组作用存在统计学差异(与模型对照组比较,P＜0.01).结论:川木香具有抑制实验性胃溃疡的形成作用,醋酸乙酯提取物可以作为川木香抑制胃溃疡形成的有效部位.     

Helicobacter pylori infection and antioxidants can modulate the genotoxic effects of heterocyclic amines in gastric mucosa cells

Helicobacter pylori (H. pylori) infection plays an important role in gastric carcinogenesis. This bacterium may induce cancer transformation and change the susceptibility of gastric mucosa cells to various exogenous dietary irritants. The aim of the study was to evaluate the influence of H. pylori infection on the reaction of the stomach cells to a genotoxic effect of heterocyclic amines (HCAs). These well-known mutagens are formed during cooking of protein-rich foods, primarily meat. Taking into account that persons consuming a mixed-western diet are exposed to these compound nearly an entire lifetime and more than half of human population is infected with H. pylori, it is important to assess the combined effect of H. pylori infection and HCAs in the context of DNA damage in gastric mucosa cells, which is a prerequisite to cancer transformation. We employed 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,8-dimethyl-imidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) because these substances are present in a great amount in cooked and fried meat. Using alkaline comet assay, we showed that the extent of the DNA damage induced by HCAs was significantly higher in H. pylori infected gastric mucosa cells than in non-infected counterparts. We did not observed any difference in the efficiency of repair of DNA lesions induced by HCAs in both type of cells. Vitamin C reduced the genotoxic effects of HCAs in H. pylori infected and non-infected gastric mucosa cells. Melatonin more effectively decreased DNA damage caused by HCAs in H. pylori infected gastric mucosa cells as compared with control. Our results suggest that H. pylori infection may influence the susceptibility of gastric mucosa cells to HCAs and dietary antioxidative substances, including vitamin C and melatonin may inhibit the genotoxic effects of HCAs on gastric mucosa cells and may reduce the risk of carcinogenesis caused by food borne mutagens and H. pylori infection.     

Protective effect of rosmarinic acid-rich trichodesma khasianum clarke leaves against ethanol-induced gastric mucosal injury in vitro and in vivo

BackgroundAlthough gastroprotective drugs have been used for peptic ulcer disease prevention and treatment, side effects have been observed. Finding a safe and effective treatment strategy is important.PurposeEdible Trichodesma khasianum (T. khasianum) Clarke leaves are considered to protect against peptic ulcers. However, scientific evidence of this effect of T. khasianum Clarke leaves remains limited.Study design/methodsIn this study, we aimed to evaluate the effect of T. khasianum Clarke leaves on ethanol-induced gastric injury and gut microbiota using RAW 264.7 cells, RGM-1 cells, and BALB/c mice, respectively.ResultThe rosmarinic acid was identified as the major component of T. khasianum Clarke leaves extracted by 80% ethanol (80EETC). The results showed that 80EETC suppressed inflammatory mediator protein levels in LPS-induced RAW 264.7 cells. Additionally, heat shock protein expression, antiapoptotic ability, and wound healing migration capability were increased by 80EETC pretreatment in RGM-1 cells with the ethanol-induced injury. Remarkably, pretreatment with 80EETC (150 mg/kg b.w.) promoted gastric mucosal healing by decreasing oxidative stress, inflammatory response, proapoptotic protein expression, and gastric mucosa damage in ethanol-induced gastric injury in mice. Crucially, no liver or kidney toxicities were observed by 80EETC oral gavage. Moreover, 80EETC increased gut microbiota diversity and short-chain fatty acid production.ConclusionOur results illustrated the remarkable gastroprotective effect by 80EETC treatment in vitro and in vivo. These findings are the first to demonstrate the powerful protective effect of T. khasianum Clarke leaves against gastric mucosal injury development.     

Effect of epiberberine from Coptis chinensis Franch on inhibition of tumor growth in MKN-45 xenograft mice

Background and purposeGastric cancer is one of the major malignancies worldwide. Epiberberine (EPI) is a major alkaloid from Coptis chinensis Franch and the antitumor property of EPI remains poorly understood.MethodThe inhibition on gastric cancer cells was observed by MTT assays and colony formation experiments. The apoptosis, cell cycle, and reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) in gastric cancer cells were analyzed by Flow cytometry. The anti-tumor effect of EPI was evaluated with the MKN-45-beraring nude mice, and the potential mechanisms were explored by RNA-seq, qPCR, siRNA silencing and western blotting.ResultsEPI inhibited the proliferation of human gastric cancer cell lines MKN-45 (harboring wild-type p53) and HGC-27 (harboring mutant p53) in a dose dependent manner. EPI induced the apoptosis and cell cycle arrest in these two cell lines, of which MKN-45 cells are more sensitive to EPI than HGC-27 cells. Further experiments indicated that EPI induced the accumulation of ROS and decreased of ΔΨm in MKN-45 cells. The significant differentially expressed genes obtained by RNA-seq were distinctly enriched in the p53 signaling pathway. The apoptosis induced by EPI in MKN-45 cells would be effectively inhibited with the treatment of p53 siRNA and p53 inhibitor PFT-α. Western blotting demonstrated that EPI diminished the expression of Bcl-2 and XIAP, and increased those of p53, Bax, p21, p27, Cytochrome C and Cleaved-caspase 3. Animal experiments confirmed that EPI significantly alleviated tumor growth in MKN-45 xenograft mice via p53/Bax pathway.ConclusionsThese data indicated that EPI could be a novel anti-tumor candidate against MKN-45-related gastric cancer via targeting p53-dependent mitochondria-associated pathway.     

四川菊科植物新记录

报道四川菊科植物分布新记录15种,即白山蒿、韦谷草白背变种、百能葳、小一点红、异羽千里光、无毛蟹甲草、黑紫橐吾、蒜叶婆罗门参、华北鸦葱、毛柄蒲儿根、三尖千里光、华麻花头、棉头风毛菊、灰白风毛菊、丽江蓟。列出这些种的标本引证、地理分布和近缘种的区别等。     

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

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