Chemical characterization of three different extracts obtained from Chelidonium majus L. (Greater celandine) with insights into their in vitro, in silico and network pharmacological properties.

Plant species C. majus, which is a very rich source of secondary metabolites, was used to obtain extracts, using a conventional extraction technique. For the extraction of bioactive molecules, three solvents were used: ethyl acetate, methanol and water, which differ from each other based on their polarity. The obtained extracts were examined in terms of chemical composition, antioxidant, enzyme inhibitory activity, and cytotoxic effects. The research results indicate that methanol was a better and more efficient extractant in the process of isolating bioactive compounds than ethyl acetate and water. The chemical composition of this solvent, i.e. its polarity, contributed the most to the extraction of alkaloids and flavonoids. The high content of total phenolic compounds in the methanol extract, as well as individual alkaloids, caused a very strong antioxidant activity, as well as a strong inhibitory power when it comes to inhibiting the excessive activity of cholinesterase and tyrosinase. Methanol and ethyl acetate extracts achieved very good cytotoxic activity against cancerous cells HGC-27 and HT-29 and did not exert a toxic effect on non-cancerous cell lines (HEK293). Extracts of plant species C. majus, especially methanol extract could be characterized as a very good starting plant material for the formulation of products intended for various branches of the food and pharmaceutical industry.

The Greater Celandine: Identification and Characterization of an Antimicrobial Peptide from Chelidonium majus.

Chelidonium majus, known as Greater Celandine, is a latex-bearing plant that has been leveraged for its anticancer and antimicrobial properties. Herein, C. majus aerial tissue is mined for the presence of antimicrobial peptides. A highly abundant cysteine-rich peptide with a length of 25 amino acids, deemed CM-AMP1, is characterized through multiple mass spectrometric approaches. Electron-activated dissociation is leveraged to differentiate between isoleucine and leucine residues and complement conventional collision-induced dissociation to gain full sequence coverage of the full-length peptide. CM-AMP1 shares little sequence similarity with any proteins in publicly available databases, highlighting the novelty of its cysteine landscape and core motif. The presence of three disulfide bonds in the native peptide confers proteolytic stability, and antimicrobial activity is greatly decreased upon the alkylation of the cysteine residues. Synthetic variants of CM-AMP1 are used to confirm the activity of the full-length sequence and the core motif. To assess the biological impact, E. coli was grown in a sublethal concentration of CM-AMP1 and quantitative proteomics was used to identify proteins produced by the bacteria under stress, ultimately suggesting a membrane lytic antimicrobial mechanism of action. This study integrates multiple analytical methods for molecular and biological characterization of a unique antimicrobial peptide identified from C. majus.

Determination of Selected Isoquinoline Alkaloids from Chelidonium majus, Mahonia aquifolium and Sanguinaria canadensis Extracts by Liquid Chromatography and Their In Vitro and In Vivo Cytotoxic Activity against Human Cancer Cells.

The search for new substances with cytotoxic activity against various cancer cells, especially cells that are very resistant to currently used chemotherapeutic agents, such as melanoma cells, is a very important scientific aspect. We investigated the cytotoxic effect of Chelidonium majus, Mahonia aquifolium and Sanguinaria canadensis extracts obtained from different parts of these plants collected at various vegetation stages on FaDu, SCC-25, MCF-7, and MDA-MB-231 cancer cells. Almost all the tested extracts showed higher cytotoxicity against these cancer cells than the anticancer drug etoposide. The highest cytotoxicity against the FaDu, SCC-25, MCF-7 and MDA-MB-231 cancer cell lines was obtained for the Sanguinaria candensis extract collected before flowering. The cytotoxicity of extracts obtained from different parts of Chelidonium majus collected at various vegetation stages was also evaluated on melanoma cells (A375, G361 and SK-MEL-3). The highest cytotoxic activity against melanoma A375 cells was observed for the Chelidonium majus root extract, with an IC50 of 12.65 µg/mL. The same extract was the most cytotoxic against SK-MEL-3 cells (IC50 = 1.93 µg/mL), while the highest cytotoxic activity against G361 cells was observed after exposure to the extract obtained from the herb of the plant. The cytotoxic activity of Chelidonium majus extracts against melanoma cells was compared with the cytotoxicity of the following anticancer drugs: etoposide, cisplatin and hydroxyurea. In most cases, the IC50 values obtained for the anticancer drugs were higher than those obtained for the Chelidonium majus extracts. The most cytotoxic extract obtained from the root of Chelidonium majus was selected for in vivo cytotoxic activity investigations using a Danio rerio larvae xenograft model. The model was applied for the first time in the in vivo investigations of the extract's anticancer potential. The application of Danio rerio larvae xenografts in cancer research is advantageous because of the transparency and ease of compound administration, the small size and the short duration and low cost of the experiments. The results obtained in the xenograft model confirmed the great effect of the investigated extract on the number of cancer cells in a living organism. Our investigations show that the investigated plant extracts exhibit very high cytotoxic activity and can be recommended for further experiments in order to additionally confirm their potential use in the treatment of various human cancers.

Relationships between Heavy Metal Concentrations in Greater Celandine (Chelidonium majus L.) Tissues and Soil in Urban Parks.

Anthropogenic ecological ecosystems create favourable conditions for the growth of the nitrophilous medicinal species Chelidonium majus in six urban parks in Southern Poland. This study focuses on the concentrations of trace elements in the soils, leaves, stems, and rhizomes of greater celandine. The soil samples were taken only in the humus horizon (A), which averaged approximately 15 cm in thickness under the clumps of Ch. majus. Regarding the reaction, the soil samples tested can be described as slightly acidic (5.6-6.8 in KCl) to alkaline (7.1-7.4 in H2O). Organic carbon content at all sites is high, ranging from 3.2% to 13.6%, while the highest total nitrogen (Nt) content is 0.664%. The average total phosphorus (Pt) content in all samples is 548.8 mg/kg (and its range is 298-940 mg/kg), such values indicating its anthropogenic origin. In terms of heavy metals, Zn has the highest content in the analysed soil samples compared to the other elements, and its range is from 394.50 mg/kg to 1363.80 mg/kg in soil. In rhizomes, Zn also has the highest values (178.7-408.3 mg/kg), whereas, in stems and leaves, it varies (from 80.6 to 227.5 and from 57.8 to 297.4 mg/kg, respectively). Spearman's rank correlation showed high correlations between the content of Pb, Zn, Cd, and As in the soil and rhizomes of Ch. majus. Despite soil contamination with Pb, Cd, and Zn, Ch. majus does not accumulate them in its tissues. However, the translocation of Hg and Cr from rhizomes to leaves was observed. The different concentrations of metals in each park result from the degree of diversity of the parent rocks on which the soil was formed.

Flower angle favors pollen export efficiency in the snowdrop Galanthus nivalis (Linnaeus, 1753) but not in the lesser celandine Ficaria verna (Huds, 1762).

Flower angle is crucially important for accurate pollination and flower protection against abiotic factors. Evolutionary factors shaping floral traits are particularly strong for bilaterally symmetric flowers because these flowers require more pollination accuracy than radially symmetrical flowers. We experimentally investigated the flower angle in the snowdrop's (Galanthus nivalis) radially symmetrical, early-blooming downward flowers. Bumblebees were able to gather significantly more pollen grains from downward flowers than from upward flowers, but female traits (fertility in the field) seem unaffected by flower angle. Similar experiments with radially symmetrical, later flowering Lesser celandine (Ficaria verna) upward flowers showed no differences in bees' abilities to gather pollen in upward vs downward-facing flowers. The downward angle of snowdrop flowers is an adaptation that increases the ability of insects to collect more pollen grains under unfavorable early spring weather conditions when pollinators are scarce.

The Activity of Chelidonium majus L. Latex and Its Components on HPV Reveal Insights into the Antiviral Molecular Mechanism.

Yellow-orange latex of Chelidonium majus L. has been used in folk medicine as a therapeutic agent against warts and other visible symptoms of human papillomavirus (HPV) infections for centuries. The observed antiviral and antitumor properties of C. majus latex are often attributed to alkaloids contained therein, but recent studies indicate that latex proteins may also play an important role in its pharmacological activities. Therefore, the aim of the study was to investigate the effect of the crude C. majus latex and its protein and alkaloid-rich fractions on different stages of the HPV replication cycle. The results showed that the latex components, such as alkaloids and proteins, decrease HPV infectivity and inhibit the expression of viral oncogenes (E6, E7) on mRNA and protein levels. However, the crude latex and its fractions do not affect the stability of structural proteins in HPV pseudovirions and they do not inhibit the virus from attaching to the cell surface. In addition, the protein fraction causes increased TNFα secretion, which may indicate the induction of an inflammatory response. These findings indicate that the antiviral properties of C. majus latex arise both from alkaloids and proteins contained therein, acting on different stages of the viral replication cycle.

Effectiveness of Volatile Natural Deep Eutectic Solvents (VNADESs) for the Green Extraction of Chelidonium majus Isoquinoline Alkaloids.

The Chelidonium majus plant is rich in biologically active isoquinoline alkaloids. These alkaline polar compounds are isolated from raw materials with the use of acidified water or methanol; next, after alkalisation of the extract, they are extracted using chloroform or dichloromethane. This procedure requires the use of toxic solvents. The present study assessed the possibility of using volatile natural deep eutectic solvents (VNADESs) for the efficient and environmentally friendly extraction of Chelidonium alkaloids. The roots and herb of the plant were subjected three times to extraction with various menthol, thymol, and camphor mixtures and with water and methanol (acidified and nonacidified). It has been shown that alkaloids can be efficiently isolated using menthol-camphor and menthol-thymol mixtures. In comparison with the extraction with acidified methanol, the use of appropriate VNADESs formulations yielded higher amounts of protopine (by 16%), chelidonine (35%), berberine (76%), chelerythrine (12%), and coptisine (180%). Sanguinarine extraction efficiency was at the same level. Additionally, the values of the contact angles of the raw materials treated with the tested solvents were assessed, and higher wetting dynamics were observed in the case of VNADESs when compared with water. These results suggest that VNADESs can be used for the efficient and environmentally friendly extraction of Chelidonium alkaloids.

Eutectic Thin-Layer Chromatography as a New Possibility for Quantification of Plant Extracts-A Case Study.

Deep eutectic solvents (DES), compared to classic ones, have interesting properties, such as the ability to solubilize compounds differing in polarity or increased dissolution of selected chemical compounds. They also offer specific interactions between the mobile and stationary phases. Those features make them promising solvents in chromatographic techniques, including the use in the separation of complicated samples. The first quantitative analysis with eutectic thin-layer chromatography (TLC) is presented in the paper. As a case study, five alkaloids from Chelidonium maius were selected as target compounds. A wide range of terpene-based DESs was investigated to develop the chromatographic system, both pure and after dilution. Moreover, a novel approach was employed to adjust polarity, involving mixing DESs differing in chromatographic properties. This procedure has proved to be effective. The best results were obtained with a 2:1 (wt/wt) mixture of DESs: camphor + phenol and menthol + limonene, with a 20% addition of methanol. The chromatographic system was validated and checked on the real sample, which made it the first applicable and operational quantitative eutectic TLC system.

Chelidonium majus Induces Apoptosis of Human Ovarian Cancer Cells via ATF3-Mediated Regulation of Foxo3a by Tip60.

Forkhead transcription factor 3a (Foxo3a) is believed to be a tumor suppressor as its inactivation leads to cell transformation and tumor development. However, further investigation is required regarding the involvement of the activating transcription factor 3 (ATF3)-mediated Tat-interactive protein 60 (Tip60)/Foxo3a pathway in cancer cell apoptosis. This study demonstrated that Chelidonium majus upregulated the expression of ATF3 and Tip60 and promoted Foxo3a nuclear translocation, ultimately increasing the level of Bcl-2-associated X protein (Bax) protein. ATF3 overexpression stimulated Tip60 expression, while ATF3 inhibition by siRNA repressed Tip60 expression. Furthermore, siRNA-mediated Tip60 inhibition significantly promoted Foxo3a phosphorylation, leading to blockade of Foxo3a translocation into the nucleus. Thus, we were able to deduce that ATF3 mediates the regulation of Foxo3a by Tip60. Moreover, siRNA-mediated Foxo3a inhibition suppressed the expression of Bax and subsequent apoptosis. Taken together, our data demonstrate that Chelidonium majus induces SKOV-3 cell death by increasing ATF3 levels and its downstream proteins Tip60 and Foxo3a. This suggests a potential therapeutic role of Chelidonium majus against ovarian cancer.

Preventive Efficiency of Chelidonium majus Ethanolic Extract Against Aflatoxin B1 Induced Neurochemical Deteriorations in Rats.

<b>Background and Objective:</b> Aflatoxins affect many species including humans and animals, therefore the present study was designed to investigate the protective effect of <i>Chelidonium majus</i> Ethanolic Extract (CMEE) on neurotoxicity induced by Aflatoxin B₁ (AFB1) in rats. <b>Materials and Methods:</b> Four groups of male Albino rats were treated orally for 28 days as follows: (1) Control group was daily given DMSO-PBS buffer (1.0 mL per rat), (2) CMEE (300 mg kg¹/day) dissolved in DMSO-PBS buffer, (3) AFB1 (80 µg kg¹/day) dissolved in DMSO-PBS buffer and (4) Received daily AFB1 (300 mg kg¹) in combination with CMEE (300 mg kg¹). <b>Results:</b> CMEE exhibits antioxidant activity <i>in vitro</i> and neuroameliorative efficiency <i>in vivo</i> as its administration in combination with AFB1 succeeded significantly in down regulating the elevated levels of inflammatory and apoptotic markers and restoring the values of neurochemical markers (AChE-ase, dopamine and serotonin) that were deteriorated by AFB1 intake. <b>Conclusion:</b> In conclusion, the neuroprotective effect of CMEE may be mediated through its antioxidant and free radical scavenging activity that proved from the data<i> </i>of ferric-reducing power ability and DPPH radical scavenging activity.

Investigation of in vitro Cytotoxicity of Chelidonium majus against Leishmania major.

One of the public health issues in the endemic areas, especially in the Middle East region would be the Leishmaniasis. The suggested cure for leishmaniasis is pentavalent antimonials. These medications have drastic side effects and the risk of relapse. On the other hand, nowadays use of herbal remedies as safe and cost-effective treatments have been increased. Therefore this study was designed to determine in vitro anti-leishmanial activity of methanol extracts of greater celandine (Chelidonium majus) against Leishmania major. Greater celandine extract was added to L. major promastigotes and intra-macrophagic amastigotes. After 24, 48 and 72 h in vitro culture the percentage of promastigotes viability was calculated by direct counting method and MTT assay. Cytotoxicity in intra-macrophagic amastigotes was evaluated by direct counting method. Viability in minimum dose and maximum dose-treated groups (1.5 and 90 µg/ml) after 24 h, was 55.52% and 36.34%, respectively. After 48 h, it was 40% and 25.26% and after 72 h, it was 62.18% and 38.45%, respectively. The half maximal inhibitory concentration (IC50) was 0.92 µg/ml, after 24 h. Cytotoxicity in intra-macrophagic amastigotes treated by 3 µg/ml dose after 24 and 48 h, was 33.23% and 50.34%, respectively. It could be concluded that greater celandine methanolic extract has in vitro cytotoxic effect on the L. major in time and dose-dependent pattern.

Synthesis and characterization of chitosan nanoparticles loaded with greater celandine (Chelidonium majus L.) essential oil as an anticancer agent on MCF-7 cell line.

Essential oils (EOs) of greater celandine (GC) roots and leaves were extracted, and gas chromatography-mass spectrometry (GC-MS) was used for analyzing them. Then they were loaded into chitosan nanoparticles (CNPs) using emulsion-ionic gelation method. CNPs loaded with greater celandine root essential oil (GCREO) and leave essential oil (GCLEO) were synthesized (size 76.5-115.3 nm) using an emulsion-ionic gelation method. Fourier Transform Infrared (FT-IR), spectroscopy, scanning electron microscope (SEM), and dynamic light scattering (DLS) were used for characterization of the formed NPs. Good encapsulation efficiency was confirmed for GCREO (62.5%) and GCLEO (69.1%) in CNPs. According to the MTT results, the synthesized NPs showed a dose-dependent effect on MCF-7 cell line. The inhibitory concentration (IC50) values for GCREO, GCLEO, CSNRs-GCREO and CNPs-GCLEO samples were 126.4, 90.2, 77.6, and 41.5 µg/mL, respectively. The highest rate of apoptosis was obtained in the CNPs-GCLEO group (63.73%). The results revealed that the cytotoxicity of CSNRs-GCREO and CNPs-GCLEO against MCF-7 cell line was significantly higher than that of their free form, implying that encapsulation of GCREO and GCLEO in CNPs is an efficient technique for improving their anti-cancer activity against MCF-7 cell line.

Chelidonium majus crude extract induces activation of peripheral blood mononuclear cells and enhances their cytotoxic effect toward HeLa cells.

The aim of the study was to examine the immunomodulatory effect of crude Chelidonium majus L ethanolic extract on ex vivo harvested peripheral blood mononuclear cells (PBMNCs). PBMNCs were isolated by density gradient centrifugation. The PBMNC cytotoxicity assay was performed with HeLa tumor cells as target cells. MTT assay was used to estimate the proliferation effect of extract and cytotoxic efficiency of treated PBMNCs. Flow cytometric analysis was used for immunophenotyping. Treatment induced moderate proliferative response, perturbation in PBMNC ratios, and the emergence of some unconventional subpopulations. The percentage ratio of double positive CD4+ and CD8+ T lymphocytes and monocytes, ratio of T and B lymphocytes expressing CD14, and percentage of NK cells expressing CD57 increased after treatment, indicating activation of PBMNC subpopulations. Cytotoxic activity against HeLa cells was enhanced. Activation of PBMNCs and enhancement of their cytotoxic effect toward HeLa cells indicate the immunostimulatory effect of Ch. majus ethanolic extract.

Combined Protein and Alkaloid Research of Chelidonium majus Latex Reveals CmMLP1 Accompanied by Alkaloids with Cytotoxic Potential to Human Cervical Carcinoma Cells.

Chelidonium majus L. is a latex-bearing plant used in traditional folk medicine to treat human papillomavirus (HPV)-caused warts, papillae, and condylomas. Its latex and extracts are rich in many low-molecular compounds and proteins, but there is little or no information on their potential interaction. We describe the isolation and identification of a novel major latex protein (CmMLP1) composed of 147 amino acids and present a model of its structure containing a conserved hydrophobic cavity with high affinity to berberine, 8-hydroxycheleritrine, and dihydroberberine. CmMLP1 and the accompanying three alkaloids were present in the eluted chromatographic fractions of latex. They decreased in vitro viability of human cervical cancer cells (HPV-negative and HPV-positive). We combined, for the first time, research on macromolecular and low-molecular-weight compounds of latex-bearing plants in contrast to other studies that investigated proteins and alkaloids separately. The observed interaction between latex protein and alkaloids may influence our knowledge on plant defense. The proposed toolbox may help in further understanding of plant disease resistance and in pharmacological research.

Functional Studies of Plant Latex as a Rich Source of Bioactive Compounds: Focus on Proteins and Alkaloids.

Latex, a sticky emulsion produced by specialized cells called laticifers, is a crucial part of a plant's defense system against herbivory and pathogens. It consists of a broad spectrum of active compounds, which are beneficial not only for plants, but for human health as well, enough to mention the use of morphine or codeine from poppy latex. Here, we reviewed latex's general role in plant physiology and the significance of particular compounds (alkaloids and proteins) to its defense system with the example of Chelidonium majus L. from the poppy family. We further attempt to present latex chemicals used so far in medicine and then focus on functional studies of proteins and other compounds with potential pharmacological activities using modern techniques such as CRISPR/Cas9 gene editing. Despite the centuries-old tradition of using latex-bearing plants in therapies, there are still a lot of promising molecules waiting to be explored.

Comparative analysis of metabolic variations, antioxidant potential and cytotoxic effects in different parts of Chelidonium majus L.

Metabolic variations, antioxidant potential and cytotoxic effects were investigated in the different plant parts like the leaf, stem, flower, pod, and root of C. majus L. using spectroscopic and chromatographic methods. Total phenolics and flavonoids were studied in the different parts of C. majus L., leaf showed higher flavonoid content (137.43 mg/g), while the pod showed the highest phenolic (23.67 mg/g) content, when compared with the stem, flower and root. In the ABTS antioxidant assay, the flower extract showed 57.94% effect, while the leaf, pod and root extract exhibited 39.10%, 36.08% and 28.88% activity, respectively. The pod and leaf extracts demonstrated the potential effect, exhibiting 45.46 and 41.61% activity, respectively, for the DPPH assay. Similar to the phosphomolybdenum assay, the flower revealed higher antioxidant activity (46.82%) than the other plant parts. The in vitro SRB assay facilitated evaluation of the cytotoxic effect against the HeLa and CaSki human cervical cancerous cells. The extract displayed dose-dependent inhibitory effect on both the cell lines. The highest cytotoxic effect was observed in the pod and flower extracts post 48 h of exposure at 1000 µg/mL. The results of C. majus L. offered new insights in the preliminary steps regarding the development of a high value product for phytomedicine applications though promising metabolic variations with antioxidant and anticancer potentials.

Effects of Abiotic Elicitors on Expression and Accumulation of Three Candidate Benzophenanthridine Alkaloids in Cultured Greater Celandine Cells.

Efforts to develop the necessary biotechnologies in Greater Celandine (Chelidonium majus L.), a leading plant resource for the development of plant-derived medicines, have been hampered by the lack of knowledge about transcriptome and metabolome regulations of its medicinal components. Therefore, this study aimed to examine the effect of abiotic elicitors, methyl jasmonate (MJ) and salicylic acid (SA), at different time courses (12, 24, 48, and 72 h), on expression and metabolome of key benzophenanthridine alkaloids (BPAs) in an optimized in vitro culture. Gene expression analysis indicated the upregulation of CFS (cheilanthifoline synthase) to 2.62, 4.85, and 7.28 times higher than the control at 12, 24, and 48 h respectively, under MJ elicitation. Besides, MJ upregulated the expression of TNMT (tetrahydroprotoberberine N-methyltransferase) to 2.79, 4.75, and 7.21 times at 12, 24, and 48 h respectively, compared to the control. Investigation of BPAs revealed a significant enhancement in the chelidonine content (9.86 µg/mg) after 72 h of MJ elicitation. Additionally, sanguinarine content increased to its highest level (3.42 µg/mg) after 24 h of MJ elicitation; however, no significant enhancement was detected in its content in shorter elicitation time courses. Generally, higher gene expression and BPAs' level was observed through longer elicitation courses (48 and 72 h). Our findings take part in improving the understanding of transcription and metabolic regulation of BPAs in cultured Greater Celandine cells.

Greater celandine (Chelidonium majus L.) for COVID-19: A twenty-case series.

In December 2019, an outbreak of coronavirus disease 2019 (COVID-19) occurred in Wuhan, China, with a rapid increase in cases worldwide. Until now, among several drugs tested, none demonstrated sufficient efficacy for its etiological treatment. Greater celandine (Chelidonium majus L.) is a well-known medicinal plant, traditionally indicated for digestive disorders and topically to remove warts. This study, performed at private offices in São Paulo and Aracaju (Brazil), describes 20 consecutive COVID-19 outpatients treated with greater celandine and their clinical evolution. The patients, aged 14-71 years (median of 41 years), were treated with Chelidonium majus 10% mother tincture, 20-30 drops three times a day for 3-12 days (median of 5 days). Clinical features were assessed during the treatment and at least until 1 week after its end. These cases were considered mild, as most COVID-19 cases. The symptoms were mainly fever, fatigue, cough, sore throat, coryza, anosmia, ageusia, and headache. Ten patients had comorbidities, such as hypertension, diabetes, and overweight. Complete or almost complete clinical improvement occurred within 1-9 days of treatment (median of 3 days). There were no adverse events. This casuistry, although small, may inspire other researchers to continue investigating Chelidonium majus as a healing treatment for COVID-19.

Anti-TMV activity and mode of action of three alkaloids isolated from Chelidonium majus.

BACKGROUND: Plant viral diseases are difficult to control and have caused serious damage to the agricultural industry. Recently, botanical biopesticides characterized by environment friendly, safe to non-target organism and not as susceptible to produce drug resistance, have exhibited great potential to be developed as antiviral agents. To screen the natural products with antiviral effect, three alkaloids possessed anti-tobacco mosaic virus (TMV) activity were isolated from Chelidonium majus and the modes of action were investigated. RESULT: The anti-TMV effect of crude extracts at 10 mg mL-1 was 51.73%. Bioassay-guided fractionation and isolation of the compounds with anti-TMV activity were performed on the methanol extract of C. majus yielding three bioactive alkaloids namely: chelerythrine (1), chelidonine (2), and sanguinarine (3). The results of bioassay showed that chelerythrine exhibited great inactivation, proliferation inhibition and protection effects against TMV at 0.5 mg mL-1 with the efficiency of 72.67%, 77.52% and 59.34%, respectively. Chelidonine at 0.1 mg mL-1 can provide 54.90% and 64.45% inhibitions on TMV through inducing resistance in two kinds of tobacco. Sanguinarine showed a weaker protection for resisting TMV in comparison to chelerythrine and chelidonine. CONCLUSION: Chelerythrine and chelidonine displayed significant inhibitions on TMV with different modes of action. These results provided important evidence that the extracts in C. majus might be a potential source of new drugs in controlling virus disease agriculturally.