Sanguinarine Attenuates Lipopolysaccharide-induced Inflammation and Apoptosis by Inhibiting the TLR4/NF-KB Pathway in H9c2 Cardiomyocytes

The inflammatory response is involved in the pathogenesis of the most common types of heart disease. Sanguinarine （SAN） has various pharmacological properties such as anti-inflammatory, antioxidant, antibacterial, antitumor, and immune-enhancing properties. However, few studies have investigated the effects of SAN on lipopolysaceharide （LPS）-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes. Therefore, in this study, H9c2 cells were co-treated with SAN and LPS, and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation. The underlying mechanism was further investigated by detecting the activation of Toll-like receptor （TLR）4/nuclear faetor-κB （NF-κB） signaling pathways. As a result, increased mRNA expression of interleukin （IL）-1β, IL-6, and TNFα induced by LPS was attenuated after SAN treatment; LPS-induced apoptosis ofHge2 cardiomyocytes and cleaved-caspase 8, 9, 3 were all significantly reduced by SAN. Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway. It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes, which may be mediated by inhibition of the TLR4/NF-κB signaling pathway. Thus, SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.

Antitumor effects of the benzophenanthridine alkaloid sanguinarine: Evidence and perspectives

Historically, natural products have represented a significant source of anticancer agents, with plant-derived drugs becoming increasingly explored. In particular, sanguinarine is a benzophenanthridine alkaloid obtained from the root of Sanguinaria canadensis, and from other poppy Fumaria species, with recognized anti-microbial, anti-oxidant and anti-inflammatory properties. Recently, increasing evidence that sanguinarine exibits anticancer potential through its capability of inducing apoptosis and/or antiproliferative effects on tumor cells, has been proved. Moreover, its antitumor seems to be due not only to its pro-apoptotic and inhibitory effects on tumor growth, but also to its antiangiogenic and anti-invasive properties. Although the precise mechanisms underlying the antitumor activity of this compound remain not fully understood, in this review we will focus on the most recent findings about the cellular and molecular pathways affected by sanguinarine, together with the rationale of its potential application in clinic. The complex of data currently available suggest the potential application of sanguinarine as an adjuvant in the therapy of cancer, but further pre-clinical studies are needed before such an antitumor strategy can be effectively translated in the clinical practice.

Spectral studies of the interaction between sanguinarine and guanosine

The interaction between sanguinarine and guanosine was investigated by using UV-vis and fluorescence spectra at pH 7.2. The binding of sanguinarine to guanosine was substantiated by the hypochromism and bathochromism in the absorption spectra and the emission quenching in fluorescence spectra. The fluorescence lifetime results, the varieties of the fluorophore absorption spectra and the decrease of the binding constant with the increasing temperature all indicate that the fluorescence quenching is static. The ratio and constant of the binding cytidine to sanguinarine are 2 and 6.44 × 10^7, respectively. The result shows that the binding of sanguinarine to guanosine is not only exothermic but also entropy-driven with △H=-8.53kJ/mol, AS = 0.12 kJ/（molK）, and AG =-44.57 kJ/mol at 298.15 K.

Stability of Sanguinarine and Chelerythrine in Macleaya cordata (Willd.) R. Br.

[Objectives]To explore the stability of sanguinarine and chelerythrine in Macleaya cordata(Willd.)R.Br.[Methods]The solubility and stability of sanguinarine and chelerythrine in seven solvents were measured by HPLC.Besides,the effects of water quality,light source,oxidant,temperature,and pH on stability were investigated.[Results]The solubility and stability of sanguinarine and chelerythrine in methanol and ethanol are good;the stability of sanguinarine and chelerythrine in distilled water and rainwater is not affected by light and is very stable,but they are unstable in tap water whether they are protected from light or not;oxidants have a great influence on the stability of sanguinarine and chelerythrine;sanguinarine and chelerythrine are stable at room temperature lower than 54℃;sanguinarine is stable in pH 2.5-7.0,while chelerythrine is stable in pH 2.5-8.0.[Conclusions]The good stability of sanguinarine and chelerythrine under specific conditions shows that they have broad development prospects and value.

Sanguinarine-Mediated Sensitization of Cervical Cancer SiHa Cells to TRAIL

Introduction: Cervical cancer is primarily caused by the human papilloma virus (HPV), which transforms normal cervical cells into cancerous cells that are highly resistant to radiation and chemotherapy. Induction of apoptosis in transformed cells is a key strategy in successfully treating HPV-induced cervical cancer. TRAIL (tumor necrosis factor related apoptosis-inducing ligand) has been shown to selectively induce apoptosis in cancer cells by binding to death receptors and activating extrinsic pathways for apoptosis. However, certain cervical cancers—such as the cultured cell line SiHa—are remarkably resistant to TRAIL. In this study, SiHa cells were sensitized to TRAIL by using sanguinarine—derived from the plant Sanguinaria Canadensis—which is known to induce oxidative stress and lead to the upregulation of receptors for TRAIL. Methods: Cultured SiHa cells were exposed to sub-lethal doses of sanguinarine in combination with TRAIL. Cell viability changes as well as the production of reactive oxygen species (ROS) were assessed. The induction of apoptosis was investigated by assays for caspase activation. Flow cytometry was performed to analyze expression of death receptors 4/5. Results: Treatment of SiHa cells with a combination of sanguinarine and TRAIL led to a significant reduction in cell viability. Significant increase in ROS was observed and caspase activation assays confirmed the induction of apoptosis. Conclusions: The observed synergistic effect of sanguinarine and TRAIL on SiHa cells is promising for the treatment of cervical, and possibly other, HPV-induced cancers. Oxidative stress caused by sanguinarine seems to play a central role in this synergy. The precise link between reactive oxygen species and the possible upregulation of death receptors needs further investigation. This knowledge will enable us to devise more effective treatments for those who suffer from this devastating disease.

Simultaneous Quantitative Determination of Nitidine, Chelerythrine and Sanguinarine Using HPTLC from Callus Extract of <i>Zanthoxylum rhetsa</i>

Nitidine, Chelerythrine and Sanguinarine, all these three alkaloids are benzophenanthridine alkaloids. Nitidine was used as an anti-HIV, anti-malarial and anti-cancer. Chelerythrine had anti-cancer and anti-inflammatory activities. Sanguinarine was widely used as an anti-plaquestic and anti-cancer. High performance thin layer chromatography (HPTLC) method was used for simultaneous quantification of Nitidine, Chelerythrine and Sanguinarine in callus extract of Zanthoxylum rhetsa by using Silica gel 60 F254 as stationary phase and ethyl acetate:methanol:water:diethylamine (30:5:2:0.5 v/v) as mobile phase at 280 nm. The linearity concentration range was 5 - 160 μg/band of each alkaloid. The Rf values of Nitidine, Chelerythrine and Sanguinarine were found to be 0.28, 0.49 and 0.73. The limit of detection and limit of quantification were found to be 0.026, 0.088 μg/spot and 0.010 and 0.033 μg/spot, 0.0104 and 0.035 μg/spot respectively for Nitidine, Chelerythrine and Sanguinarine. HPTLC method was developed and validated according to ICH guidelines for simultaneous estimation of Nitidine, Chelerythrine and Sanguinarine and proved to be simple, specific, accurate, robust and rapid.

Systematic understanding of the underlying target and mechanism of sanguinarine against nasopharyngeal carcinoma through a networkpharmacology approach

Background:Nasopharyngeal carcinoma is a malignant tumor,well known as a cancer type characterized by regional specificity,especially in Southern China.The network pharmacology is an emerging discipline developed in recent years,which has been effectively used to predict the potential therapeutic compounds against disease focusing on the possible therapeutic targets and mechanisms.Sanguinarine,a traditional natural plant-derived phenanthridine alkaloid,has been reported to have a wide variety of pharmacological activities for decades.Methods:In the current study,using the comprehensive network pharmacological method,the potential drug targets of sanguinarine against nasopharyngeal carcinoma were successfully predicted,and verified by molecular docking.The underlying pharmacological mechanism was initially unraveled.Results:Totally,38 potential common targets were confirmed from these potential nasopharyngeal carcinoma therapeutic targets and pharmacological targets of sanguinarine.Their enrichment analyses of GO functions show that protein serine/threonine kinase activity,histone kinase activity,integrin binding,protein tyrosine kinase activity,and cell adhesion molecule binding were top listed.KEGG functional enrichment analysis indicates that the potential pathways are mainly involved into PI3K-Akt signaling pathway.The"drug-target-disease-pathway"network model diagram points out the key genes containing MAPK10,MAPK14,JAK2,BRAF,GSK3B,MET,HSP90AA1,SRC,et al..According to the results of molecular docking,it was further verified that sanguinarine has strong binding ability with MAPK10 and MAPK14.Conclusion:Taken together,this study would provide a clarified theoretical basis for the subsequent wet laboratory research focusing on sanguinarine against nasopharyngeal carcinoma.

Binding of nucleosides with the cytotoxic plant alkaloid sanguinarine:Spectroscopic and thermodynamic studies

In spite of a large number of studies of the interaction of the cytotoxic plant alkaloid sanguinarine(SAN) with nucleic acids,the anticancer mechanism of SAN is still not clear.In contrast to the large number of studies of the interaction mechanism of SAN with DNA,there have been relatively few studies of the interaction of SAN with nucleosides.In this work,the interaction of SAN with three nucleosides-thymidine(T),uridine(U),and adenosine(A)-was investigated using a combination of conventional fluorescence and UV-vis spectroscopic techniques;thermodynamic calculations were also carried out at physiological pH 7.2.The binding processes of SAN with the different nucleosides were characterized by hypochromic and bathochromic effects in the absorption spectra of SAN and by the quenching of the fluorescence intensity of SAN.The measurements of fluorescence lifetime,the variations of the absorption spectra of the fluorophore,and the dependence of the quenching on the temperature indicated that the fluorescence quenching is static.The Stern-Volmer plot is nonlinear and approximately quadratic showing that,in this process,one SAN molecule can bind with two nucleoside molecules.These studies,together with our earlier studies of the binding of SAN with cytidine(C) and guanosine(G),showed that the binding constants of SAN with the five nucleosides at T = 308.15,318.15,and 328.15 K decreased in the order C > G > T > U > A and at T = 298.15 K decreased in the order G > C > T > U > A,and that the binding of SAN with the various nucleosides is not only slightly exothermic but also entropy-driven.All these results together with fluorescence quenching experiments advance good evidence concerning the interaction of SAN with various nucleosides.Such studies of the interaction mechanism of alkaloids with DNA may promote the development of new drugs.

Sanguinarine suppresses cell proliferation, migration and invasion in nasopharyngeal carcinoma via inhibiting mTOR signaling

OBJECTIVE: To confirm the anti-NPC effect of sanguinarine(SA) through a series of wet experiments.METHODS: NPC cell viability was determined by proliferation experiment. Cell clone formation experiment, cell scratch test, transwell migration and invasion experiment and flow cytometry-based cell apoptosis assay were further performed. In addition, Western blotting was performed to investigate the cell signaling pathway. All the relevant experimental data were statistically processed using SPSS 16.0 software.RESULTS: The results showed that sanguinarine represented a time and dose dependent inhibition effects on NPC cell proliferation including the low differentiated CNE2 cells and high metastatic 5-8F cells, along with the cell cloning ability reduction. In addition, sanguinarine has a certain inhibitory effect on the invasion and migration of NPC cells. Mechanistically, sanguinarine displayed the anti-NPC effects mainly involved into the suppression of m TOR signaling and cell apoptosis, which is closely associated with the tumor growth and metastatic malignancy. CONCLUSIONS: Collectively, we discover that sanguinarine is a new high-efficiency anti-NPC monomer of Chinese medicine, with a value for the follow-up preclinical research.

Safety Evaluation of a Standardized <i>Macleaya cordata</i>Extract in a Ninety Day Feeding Study in Weaned Piglets

The aim of this study was to assess the effects of Sangrovit?, a standardized preparation of Macleaya cordata extract (MCE), on the health status and performance in weaned piglets. A total of 128 weaned piglets were randomly divided into four groups (n = 32, 16/sex/group) and fed either a control feed (T1) or the control feed supplemented with 100 mg/kg (T2), 500 mg/kg (T3), or 1000 mg/kg (T4) Sangrovit? (resulting in 0, 3.5, 17.5, 35 mg MCE/kg feed, respectively) for ninety days. The parameters for growth and health status were evaluated during the trial and blood was collected on Day 0 and Day 91 for hematology and biochemistry analysis. After the animals were sacrificed, the weight of the major organs and tissues was measured and histopathological examination on the organs from 8 animals in each group (4/sex/group) was performed. The results showed no statistically significant differences in live weight, feed intake, and average daily weight gain between the treatment and control groups. The feed containing 100, 500, or 1000 mg/kg Sangrovit? was well tolerated by piglets, with no adverse effects noted during the feeding period or in the histopathological results. The majority of the hematological and blood biochemistry parameters displayed no significant changes between the control and treatment groups. Compared to the control group, some hematological parameters including MCV, platelets concentration, and APTT were modified in some but not all treatment groups. Some significant changes in the blood biochemistry parameters were found in the treatment groups including levels in bile acid, haptoglobin, total serum protein, and GTT, as well as creatine kinase activity, but those changes were not toxicologically significant because no observed changes were seen in other similar biomarkers. In sum, this study demonstrated that the addition of Sangrovit? to swine feed at up to 1000 mg/kg does not result in any toxicological effects to the health status and growth of weaned piglets.

Residue Study for a Standardized <i>Macleaya cordata</i>Extract in Growing-Finishing Swine

The aim of this study was to assess the effects of Sangrovit&reg;, a standardized preparation of Macleaya cordata extract (MCE), on the health status and retained residues in growing-finishing swine. A total of twelve growing-finishing swine (n = 6 for each group) were randomly divided into two groups and fed either a control feed or the control feed supplemented with 100 mg/kg Sangrovit&reg;(3.5 mg/kg MCE) for 28 days. The parameters for growth and health status were evaluated during the trial and after which the animals were slaughtered. Residual levels of MCE in swine organs and tissues were determined by measuring sanguinarine and chelerythrine levels by LC-MS/MS. The results showed no statistically significant differences in live weight, feed intake and average daily gain between the treatment and control groups. The feed supplemented with 100 mg/kg Sangrovit&reg;was well tolerated by the swine, with no adverse effects noted during the feeding period or in the necropsy results. Residue analysis indicated that levels of sanguinarine or chelerythrine were under the limit of detection in all the examined tissues and organs from the treated swine. This study demonstrates that Sangrovit&reg;, a standardized preparation of MCE when fed to growing-finishing swine for 28 days, at the level of 100 mg/kg in feed, does not result in sanguinarine or chelerythrine residues in the organs or tissues.

The Genome of Medicinal Plant Macleaya cordata Provides New Insights into Benzylisoquinoline Alkaloids Metabolism

The overuse of antibiotics in animal agriculture and medicine has caused a series of potential threats to public health. Macleaya cordata is a medicinal plant species from the Papaveraceae family, providing a safe resource for the manufacture of antimicrobial feed additive for livestock. The active constituents from M. cordata are known to include benzylisoquinoline alkaloids （BIAs） such as sanguinarine （SAN） and chelerythrine （CHE）, but their metabolic pathways have yet to be studied in this non-model plant. The active biosynthesis of SAN and CHE in M. cordata was first examined and confirmed by feeding ^13C-labeled tyrosine. To gain further insights, we de novo sequenced the whole genome of M. cordata, the first to be sequenced from the Papaveraceae family. The M. cordata genome covering 378 Mb encodes 22,328 predicted protein-coding genes with 43.5% being transposable elements. As a member of basal eudicot, M. cordata genome lacks the paleohexaploidy event that occurred in almost all eudicots. From the genomics data, a complete set of 16 metabolic genes for SAN and CHE biosynthesis was retrieved, and 14 of their biochemical activities were validated. These genomics and metabolic data show the conserved BIA metabolic pathways in M. cordata and provide the knowledge foundation for future productions of SAN and CHE by crop improvement or microbial pathway reconstruction.