The Experts below are selected from a list of 174 Experts worldwide ranked by ideXlab platform
Zhengtao Wang - One of the best experts on this subject based on the ideXlab platform.
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mass spectrometry directed analysis and purification of pyrrolizidine alkaloid cis trans isomers in gynura japonica
IEEE Journal of Solid-state Circuits, 2014Co-Authors: Lianxiang Fang, Aizhen Xiong, Xiao Yang, Wenzhi Cheng, Li Yang, Zhengtao WangAbstract:Pyrrolizidine alkaloids are highly hepatotoxic natural chemicals that produce irreversible chronic and acute hepatotoxic effects on human beings. Purification of large amounts of pyrrolizidine alkaloids is necessary for toxicity studies. In this study, an efficient method for targeted analysis and purification of pyrrolizidine alkaloid cis/trans isomers from herbal materials was developed for the first time. Targeted analysis of the hepatotoxic pyrrolizidine alkaloids was performed by liquid chromatography with tandem mass spectrometry (precursor ion scan and daughter ion scan), and the purification of pyrrolizidine alkaloids was achieved with a mass-directed auto purification system. The extraction and preparative liquid chromatography conditions were optimized. The developed method was applied to analysis of Gynura japonica (Thunb.) Juel., a herbal medicine traditionally used for detumescence and relieving pain but is potentially hepatotoxic as it contains pyrrolizidine alkaloids. Twelve pyrrolizidine alkaloids (six cis/trans isomer pairs) were identified with reference compounds or characterized by liquid chromatography with tandem mass spectrometry, and five individual pyrrolizidine alkaloids, including (E)-Seneciphylline, Seneciphylline, integerrimine, senecionine, and seneciphyllinine, were prepared from G. japonica roots with high efficiency. The results of this work provide a new technique for the preparation of large amounts of pyrrolizidine alkaloid reference substances, which will also benefit toxicological studies of pyrrolizidine alkaloids and treatments for pyrrolizidine alkaloid-induced toxicity.
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uplc ms based metabolomics study on senecio scandens and s vulgaris an approach for the differentiation of two senecio herbs with similar morphology but different toxicity
Metabolomics, 2012Co-Authors: Aizhen Xiong, Li Yang, Zhengtao Wang, Xuejing Yang, Changhong Wang, Lili Ji, Z T Wang, Ying Chen, Xiuli WangAbstract:Pyrrolizidine alkaloids show significant hepatotoxicity as they can bind to DNA or proteins after being activated in liver. Senecio vulgaris L., like many Compositae herbs containing pyrrolizidine alkaloids, was reported to have great hepatotoxicity. However, Senicio scandens Buch.-Ham., from the same genus, which was also used as a herb and documented in China Pharmacopoeia published in 2010, hardly showed any side effects or relevant toxicity. In the present study, we conducted the metabolomics study using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) to obtain the different metabolic profiles of the two Senecio herbs. In addition, principle component analysis (PCA) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) were introduced for the multivariate analysis, and MS/MS was applied to the identification of target alkaloid markers which contributed most to the established models. As a result, ten pyrrolizidine alkaloids, including adonifoline, senecionine, senecionine N-oxide, retrorsine, retrorsine N-oxide and Seneciphylline, were selected and identified. Among them, adonifoline was found to be a specific marker for S. scandens while senecionine and its N-oxidative were characteristic markers for S. vulgaris. Furthermore, the hepatotoxicity studies in vivo and in vitro showed that senecionine had more potent toxicity (LD50, 57.3 mg/kg; IC50, 5.41 μM) than that of adonifonine (LD50, 163.3 mg/kg; IC50, 49.91 μM). Taken together, the present study provides not only better understanding of the different toxicity between the two Senecio herbs containing pyrrolizidine alkaloids but also a reference method, which can be applied to other genetically closed species with similar morphology but different toxicity.
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authentication of senecio scandens and s vulgaris based on the comprehensive secondary metabolic patterns gained by uplc dad esi ms
Journal of Pharmaceutical and Biomedical Analysis, 2011Co-Authors: Xuejing Yang, Aizhen Xiong, Li Yang, Zhengtao WangAbstract:Abstract A secondary metabolic pattern using ultra-performance liquid chromatography (UPLC)–DAD/ESI-MS was constructed to gain chemical information for authentication of Senecio scandens (SS) and Senecio vulgaris (SV), the two representative species containing hepatotoxic pyrrolizidine alkaloids (HPAs). The metabolic pattern showed three groups of bioactive constituents: phenolic/aromatic acids, flavonoid glycosides and the HPAs. 47 peaks were identified including 19 phenolic/aromatic acids, 10 flavonoid glycosides and 18 PAs by direct comparison with the available reference compounds or deduced from the UV absorption and their ESI-MS fragmentation patterns. The two species could be authenticated diagnostically by their metabolic profiling of the three chromatographic fingerprints. Although both SS and SV contain PAs as the characteristic constituents, only 2 PAs, adonifoline and adonifoline N -oxide were detected in SS, while other 16 PAs were detected in SV, including the highly toxic senecionine, retrorsine, Seneciphylline and their corresponding N -oxides. The concentration of PAs in SV is also higher than that in SS. The number and concentration of the phenolic compounds in SS were higher than in SV. Jacaranone derivatives were only detected in SS and jacaranone ethyl ester was detected as the predominant constituent. In the fingerprint of the n -butanol extracts, 10 quercetin and kaempferol glycosides derivatives were detected. 9 were found in SS and only 2 in SV. PAs, jacaranone derivatives and flavonoid glycosides can serve as the metabolic markers to distinguish the Senecio plants from each other, and provide evidence for their clinical application in the consideration of safety and efficacy.
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authentication of senecio scandens and s vulgaris based on the comprehensive secondary metabolic patterns gained by uplc dad esi ms
Journal of Pharmaceutical and Biomedical Analysis, 2011Co-Authors: Xuejing Yang, Aizhen Xiong, Li Yang, Zhengtao Wang, Dingxiang LiAbstract:Abstract A secondary metabolic pattern using ultra-performance liquid chromatography (UPLC)–DAD/ESI-MS was constructed to gain chemical information for authentication of Senecio scandens (SS) and Senecio vulgaris (SV), the two representative species containing hepatotoxic pyrrolizidine alkaloids (HPAs). The metabolic pattern showed three groups of bioactive constituents: phenolic/aromatic acids, flavonoid glycosides and the HPAs. 47 peaks were identified including 19 phenolic/aromatic acids, 10 flavonoid glycosides and 18 PAs by direct comparison with the available reference compounds or deduced from the UV absorption and their ESI-MS fragmentation patterns. The two species could be authenticated diagnostically by their metabolic profiling of the three chromatographic fingerprints. Although both SS and SV contain PAs as the characteristic constituents, only 2 PAs, adonifoline and adonifoline N -oxide were detected in SS, while other 16 PAs were detected in SV, including the highly toxic senecionine, retrorsine, Seneciphylline and their corresponding N -oxides. The concentration of PAs in SV is also higher than that in SS. The number and concentration of the phenolic compounds in SS were higher than in SV. Jacaranone derivatives were only detected in SS and jacaranone ethyl ester was detected as the predominant constituent. In the fingerprint of the n -butanol extracts, 10 quercetin and kaempferol glycosides derivatives were detected. 9 were found in SS and only 2 in SV. PAs, jacaranone derivatives and flavonoid glycosides can serve as the metabolic markers to distinguish the Senecio plants from each other, and provide evidence for their clinical application in the consideration of safety and efficacy.
Ge Lin - One of the best experts on this subject based on the ideXlab platform.
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Pyrrolizidine Alkaloid-Induced Hepatotoxicity Associated with the Formation of Reactive Metabolite-Derived Pyrrole–Protein Adducts
'MDPI AG', 2021Co-Authors: Wood Yee Chan, Ge LinAbstract:Pyrrolizidine alkaloids (PAs) with 1,2-unsaturated necine base are hepatotoxic phytotoxins. Acute PA intoxication is initiated by the formation of adducts between PA-derived reactive pyrrolic metabolites with cellular proteins. The present study aimed to investigate the correlation between the formation of hepatic pyrrole–protein adducts and occurrence of PA-induced liver injury (PA-ILI), and to further explore the use of such adducts for rapidly screening the hepatotoxic potency of natural products which contain PAs. Aqueous extracts of Crotalaria sessiliflora (containing one PA: monocrotaline) and Gynura japonica (containing two PAs: senecionine and Seneciphylline) were orally administered to rats at different doses for 24 h to investigate PA-ILI. Serum alanine aminotransferase (ALT) activity, hepatic glutathione (GSH) level, and liver histological changes of the treated rats were evaluated to assess the severity of PA-ILI. The levels of pyrrole–protein adducts formed in the rats’ livers were determined by a well-established spectrophotometric method. The biological and histological results showed a dose-dependent hepatotoxicity with significantly different toxic severity among groups of rats treated with herbal extracts containing different PAs. Both serum ALT activity and the amount of hepatic pyrrole–protein adducts increased in a dose-dependent manner. Moreover, the elevation of ALT activity correlated well with the formation of hepatic pyrrole–protein adducts, regardless of the structures of different PAs. The findings revealed that the formation of hepatic pyrrole–protein adducts—which directly correlated with the elevation of serum ALT activity—was a common insult leading to PA-ILI, suggesting a potential for using pyrrole–protein adducts to screen hepatotoxicity and rank PA-containing natural products, which generally contain multiple PAs with different structures
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assessment of pyrrolizidine alkaloid induced toxicity in an in vitro screening model
Journal of Ethnopharmacology, 2013Co-Authors: Winnie Lai Ting Kan, Ge LinAbstract:Abstract Ethnopharmacological relevance Pyrrolizidine alkaloids (PAs) are a group of heterocyclic phytotoxins present in a wide range of plants. The consumption of PA-containing medicinal herbs or PA-contaminated foodstuffs has long been reported to cause human hepatotoxicity. However, the degrees of hepatotoxicity of different PAs are unknown, which makes it difficult to determine a universal threshold of toxic dose of individual PAs for safe regulation of PA-containing natural products. The aim of the present study is to develop a simple and convenient in vitro model to assess the hepatotoxicity of different PAs. Material and methods Six common cytotoxicity assays were used to evaluate the hepatotoxicity of different PAs in human hepatocellular carcinoma HepG2 cells. Results The combination of MTT and bromodeoxyuridine incorporation (BrdU) assays demonstrated to be a suitable method to evaluate the toxic potencies of various PAs in HepG2 cells, and the results indicated that otonecine-type PA (clivorine: IC 20 =0.013±0.004 mM (MTT), 0.066±0.031 mM (BrdU)) exhibited significantly higher cytotoxic and anti-proliferative effects than retronecine-type PA (retrorsine: IC 20 =0.27±0.07 mM (MTT), 0.19±0.03 mM (BrdU)). While as expected, the known less toxic platyphylline-type PA (platyphylline: IC 20 =0.85±0.11 mM (MTT), 1.01±0.40 mM (BrdU)) exhibited significantly less toxicity. The different cytotoxic and anti-proliferative potencies of various PAs in the same retronecine-type could also be discriminated by using the combined MTT and BrdU assays. In addition, the developed assays were further utilized to test alkaloid extract of Gynura segetum , a senecionine and Seneciphylline-containing herb, the overall cytotoxicity of two PAs in the extract was comparable to that of these two PAs tested individually. Conclusion Using the developed in vitro model, the cytotoxicity of different PAs and the extract of a PA-containing herb were investigated in parallel in one system, and their different hepatotoxic potencies were determined and directly compared for the first time. The results suggested that the developed model has a great potential to be applied for the quick screening of the toxicity of PAs and PA-containing natural products.
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definitive diagnosis of hepatic sinusoidal obstruction syndrome induced by pyrrolizidine alkaloids
Journal of Digestive Diseases, 2012Co-Authors: Hong Gao, Ji Yao Wang, Shun Cai Zhang, Ge LinAbstract:OBJECTIVE: Hepatic sinusoidal obstruction syndrome (HSOS) induced by a Chinese medicinal herb Tusanqi is increasingly being reported in recent years. The aim of the study was to investigate the possibility of using blood pyrrole-protein adducts test as a confirmatory diagnostic method. METHODS: Patients with HSOS according to international diagnostic criteria associated with Tusanqi from January 2006 to August 2010 in Zhongshan Hospital Fudan University were included and clinical features were collected. Pyrrole-protein adducts in blood sample were determined with ultra performance liquid chromatography-mass spectrometry (UPLC-MS) while pyrrolizidine alkaloids (PAs) in available herbal preparations were analyzed by high performance liquid chromatography-ultraviolet (HPLC-UV). RESULTS: Five patients (age 41–72 years, median age 54 years, all women) were included. Ascites (5/5), jaundice (5/5) and hepatomegaly (4/5) were common manifestations. The imaging features were diffused, patchy hepatic enhancement, periportal edema and ascites. Pathology ascertained that blood flow was obstructive at the site of sinusoid. PAs (Seneionine and Seneciphylline) were identified in all the three available herbal preparations ingested by the HSOS patients. Pyrrole-protein adducts were unequivocally found in all the five blood samples. Two patients recovered, two developed chronic illness and one died due to liver failure and hepatic encephalopathy. CONCLUSIONS: The detection of blood pyrrole-protein adducts using a UPLC-MS approach is a specific, reliable, unambiguous and confirmatory test for HSOS induced by PA, and should be used together with the conventional HSOS clinical diagnostic criteria for the definitive diagnosis of PA-induced HSOS.
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hepatic sinusoidal obstruction syndrome associated with consumption of gynura segetum
Journal of Hepatology, 2011Co-Authors: Ge Lin, Ji Yao Wang, Hong Gao, Fan Zhang, Huali Wang, Yan Zhou, Jiang ZhengAbstract:Background & Aims One major cause of hepatic sinusoidal obstruction syndrome (HSOS) is the intake of pyrrolizidine alkaloid (PA)-containing products. Over 8000 PA-induced HSOS cases have been reported worldwide and at least 51 among them were suspected to be attributed to exposure to the Chinese medicine ‘Tusanqi'. PA-induced hepatotoxicity involves cytochrome P450-mediated metabolic activation of PAs to electrophilic pyrrolic metabolites which react with macromolecules, such as proteins. However, no studies have found such protein adduction in HSOS patients. We report one HSOS case confirmed by liver biopsy, where the patient claimed taking ‘Tusanqi' as self-medication. Methods The herb was analyzed by HPLC–MS, and its induced hepatotoxicity in rats was assessed by monitoring the alteration of serum ALT level and liver morphology. Blood pyrrole–protein adducts were determined by UPLC–MS. Results The herb the patient consumed was identified as Gynura segetum , an erroneous substitute of non-PA-containing Sedum aizoon , called ‘Tusanqi'. Hepatotoxic PAs senecionine and Seneciphylline were detected in G. segetum . In the PA-exposed patient, serum pyrrole–protein adducts were detected by a newly developed analytical approach. The animal study showed a good correlation of liver injury with the ingestion of G. segetum . Conclusions For the first time, serum pyrrole–protein adducts were unequivocally detected in a PA-induced HSOS patient, and such adducts show a potential to be developed as a biomarker for the assessment of PA-induced HSOS. Similar to the well-known case of aristolochic acid-poisoning, the observed HSOS was confirmed to arise from the consumption of PA-containing G. segetum , an erroneous substitute of non-PA-containing S. aizoon .
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identification of five hepatotoxic pyrrolizidine alkaloids in a commonly used traditional chinese medicinal herb herba senecionis scandentis qianliguang
Rapid Communications in Mass Spectrometry, 2008Co-Authors: Ge Lin, Chileung Chan, Zhihong Jiang, Zhongzhen ZhaoAbstract:Senecio scandens Buch.-Ham is a plant source for a commonly used traditional Chinese medicinal (TCM) herb Qianliguang. A TCM herbal proprietary product containing Qianliguang as the major herb for the treatment of sinusitis has been used in China for several decades, and has also been exported to other regions and countries worldwide. In the present study, the aqueous extract of S. scandens collected in the Shanxi Province of China was determined, for the first time, to contain hepatotoxic and tumorigenic pyrrolizidine alkaloids (PAs) by using high-performance liquid chromatography/mass spectrometric (HPLC/MS) analysis in various scanning modes. A total of nine toxic and two non-toxic PAs were detected in the aqueous extract of S. scandens, of which six PAs, namely neoplatyphylline, senecionine, senecionine N-oxide, Seneciphylline, Seneciphylline N-oxide and senkirkine, were unequivocally characterized, while other PAs were tentatively assigned as jacobine, jacozine N-oxide (or erucifoline N-oxide), 7-tigloylplatynecine, usaramine and an isomer of yamataimine. The estimated total content of toxic PAs in S. scandens was 10.82 microg/g herb, which was significantly higher than that (> or =1 microg/g herb) recommended by Belgium and Germany not to be used clinically. Among the PAs definitively identified, senecionine, Seneciphylline, and senkirkine are known tumorigens capable of inducing liver tumors in experimental animals, while Seneciphylline N-oxide and senecionine N-oxide are probably tumorigenic due to their potential conversion into Seneciphylline and senecionine via metabolic reduction in the body. Thus, the current finding of the presence of toxic/tumorigenic PAs in S. scandens challenges the safety of using this TCM herb and its proprietary products.
Kurt Hostettmann - One of the best experts on this subject based on the ideXlab platform.
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determination of pyrrolizidine alkaloids in senecio species by liquid chromatography thermospray mass spectrometry and liquid chromatography nuclear magnetic resonance spectroscopy
Planta Medica, 1999Co-Authors: Karine Ndjoko, Jeanluc Wolfender, Erhard Roder, Kurt HostettmannAbstract:A new method using direct on-line coupling between HPLC, MS and (1 )H-NMR was developed for the detection and identification of pyrrolizidine alkaloids (PAs) in crude extracts of Senecio species. The PAs present in the extracts were separated on a C-18 reversed-phase column with an alkaline acetonitrile-water gradient. Molecular weight information of each peak was obtained by LC/MS and specific fragments were recorded by complementary MS/MS experiments. In order to distinguish isomeric structures of PAs, complementary LC/ (1)H-NMR analyses were performed in both on-flow and stop-flow modes with alkaline acetonitrile-D (2)O and methanol-D (2)O as mobile phases. This approach led to the identification of the known PAs of S. vulgaris; retrorsine, Seneciphylline, and senecionine. The method was also applied for the analysis of African Senecio species: S. mariettae and S. venosus. Retrorsine was identified as the main PA in both species. Detection of the PAs in small amounts was achieved by LC/MS. Higher amounts of extracts (mg) had to be injected to obtain good quality on-flow LC/ (1)H-NMR.
Aizhen Xiong - One of the best experts on this subject based on the ideXlab platform.
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mass spectrometry directed analysis and purification of pyrrolizidine alkaloid cis trans isomers in gynura japonica
IEEE Journal of Solid-state Circuits, 2014Co-Authors: Lianxiang Fang, Aizhen Xiong, Xiao Yang, Wenzhi Cheng, Li Yang, Zhengtao WangAbstract:Pyrrolizidine alkaloids are highly hepatotoxic natural chemicals that produce irreversible chronic and acute hepatotoxic effects on human beings. Purification of large amounts of pyrrolizidine alkaloids is necessary for toxicity studies. In this study, an efficient method for targeted analysis and purification of pyrrolizidine alkaloid cis/trans isomers from herbal materials was developed for the first time. Targeted analysis of the hepatotoxic pyrrolizidine alkaloids was performed by liquid chromatography with tandem mass spectrometry (precursor ion scan and daughter ion scan), and the purification of pyrrolizidine alkaloids was achieved with a mass-directed auto purification system. The extraction and preparative liquid chromatography conditions were optimized. The developed method was applied to analysis of Gynura japonica (Thunb.) Juel., a herbal medicine traditionally used for detumescence and relieving pain but is potentially hepatotoxic as it contains pyrrolizidine alkaloids. Twelve pyrrolizidine alkaloids (six cis/trans isomer pairs) were identified with reference compounds or characterized by liquid chromatography with tandem mass spectrometry, and five individual pyrrolizidine alkaloids, including (E)-Seneciphylline, Seneciphylline, integerrimine, senecionine, and seneciphyllinine, were prepared from G. japonica roots with high efficiency. The results of this work provide a new technique for the preparation of large amounts of pyrrolizidine alkaloid reference substances, which will also benefit toxicological studies of pyrrolizidine alkaloids and treatments for pyrrolizidine alkaloid-induced toxicity.
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uplc ms based metabolomics study on senecio scandens and s vulgaris an approach for the differentiation of two senecio herbs with similar morphology but different toxicity
Metabolomics, 2012Co-Authors: Aizhen Xiong, Li Yang, Zhengtao Wang, Xuejing Yang, Changhong Wang, Lili Ji, Z T Wang, Ying Chen, Xiuli WangAbstract:Pyrrolizidine alkaloids show significant hepatotoxicity as they can bind to DNA or proteins after being activated in liver. Senecio vulgaris L., like many Compositae herbs containing pyrrolizidine alkaloids, was reported to have great hepatotoxicity. However, Senicio scandens Buch.-Ham., from the same genus, which was also used as a herb and documented in China Pharmacopoeia published in 2010, hardly showed any side effects or relevant toxicity. In the present study, we conducted the metabolomics study using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) to obtain the different metabolic profiles of the two Senecio herbs. In addition, principle component analysis (PCA) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) were introduced for the multivariate analysis, and MS/MS was applied to the identification of target alkaloid markers which contributed most to the established models. As a result, ten pyrrolizidine alkaloids, including adonifoline, senecionine, senecionine N-oxide, retrorsine, retrorsine N-oxide and Seneciphylline, were selected and identified. Among them, adonifoline was found to be a specific marker for S. scandens while senecionine and its N-oxidative were characteristic markers for S. vulgaris. Furthermore, the hepatotoxicity studies in vivo and in vitro showed that senecionine had more potent toxicity (LD50, 57.3 mg/kg; IC50, 5.41 μM) than that of adonifonine (LD50, 163.3 mg/kg; IC50, 49.91 μM). Taken together, the present study provides not only better understanding of the different toxicity between the two Senecio herbs containing pyrrolizidine alkaloids but also a reference method, which can be applied to other genetically closed species with similar morphology but different toxicity.
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authentication of senecio scandens and s vulgaris based on the comprehensive secondary metabolic patterns gained by uplc dad esi ms
Journal of Pharmaceutical and Biomedical Analysis, 2011Co-Authors: Xuejing Yang, Aizhen Xiong, Li Yang, Zhengtao WangAbstract:Abstract A secondary metabolic pattern using ultra-performance liquid chromatography (UPLC)–DAD/ESI-MS was constructed to gain chemical information for authentication of Senecio scandens (SS) and Senecio vulgaris (SV), the two representative species containing hepatotoxic pyrrolizidine alkaloids (HPAs). The metabolic pattern showed three groups of bioactive constituents: phenolic/aromatic acids, flavonoid glycosides and the HPAs. 47 peaks were identified including 19 phenolic/aromatic acids, 10 flavonoid glycosides and 18 PAs by direct comparison with the available reference compounds or deduced from the UV absorption and their ESI-MS fragmentation patterns. The two species could be authenticated diagnostically by their metabolic profiling of the three chromatographic fingerprints. Although both SS and SV contain PAs as the characteristic constituents, only 2 PAs, adonifoline and adonifoline N -oxide were detected in SS, while other 16 PAs were detected in SV, including the highly toxic senecionine, retrorsine, Seneciphylline and their corresponding N -oxides. The concentration of PAs in SV is also higher than that in SS. The number and concentration of the phenolic compounds in SS were higher than in SV. Jacaranone derivatives were only detected in SS and jacaranone ethyl ester was detected as the predominant constituent. In the fingerprint of the n -butanol extracts, 10 quercetin and kaempferol glycosides derivatives were detected. 9 were found in SS and only 2 in SV. PAs, jacaranone derivatives and flavonoid glycosides can serve as the metabolic markers to distinguish the Senecio plants from each other, and provide evidence for their clinical application in the consideration of safety and efficacy.
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authentication of senecio scandens and s vulgaris based on the comprehensive secondary metabolic patterns gained by uplc dad esi ms
Journal of Pharmaceutical and Biomedical Analysis, 2011Co-Authors: Xuejing Yang, Aizhen Xiong, Li Yang, Zhengtao Wang, Dingxiang LiAbstract:Abstract A secondary metabolic pattern using ultra-performance liquid chromatography (UPLC)–DAD/ESI-MS was constructed to gain chemical information for authentication of Senecio scandens (SS) and Senecio vulgaris (SV), the two representative species containing hepatotoxic pyrrolizidine alkaloids (HPAs). The metabolic pattern showed three groups of bioactive constituents: phenolic/aromatic acids, flavonoid glycosides and the HPAs. 47 peaks were identified including 19 phenolic/aromatic acids, 10 flavonoid glycosides and 18 PAs by direct comparison with the available reference compounds or deduced from the UV absorption and their ESI-MS fragmentation patterns. The two species could be authenticated diagnostically by their metabolic profiling of the three chromatographic fingerprints. Although both SS and SV contain PAs as the characteristic constituents, only 2 PAs, adonifoline and adonifoline N -oxide were detected in SS, while other 16 PAs were detected in SV, including the highly toxic senecionine, retrorsine, Seneciphylline and their corresponding N -oxides. The concentration of PAs in SV is also higher than that in SS. The number and concentration of the phenolic compounds in SS were higher than in SV. Jacaranone derivatives were only detected in SS and jacaranone ethyl ester was detected as the predominant constituent. In the fingerprint of the n -butanol extracts, 10 quercetin and kaempferol glycosides derivatives were detected. 9 were found in SS and only 2 in SV. PAs, jacaranone derivatives and flavonoid glycosides can serve as the metabolic markers to distinguish the Senecio plants from each other, and provide evidence for their clinical application in the consideration of safety and efficacy.
Hao Liang - One of the best experts on this subject based on the ideXlab platform.
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differential induction of apoptosis and autophagy by pyrrolizidine alkaloid clivorine in human hepatoma huh 7 5 cells and its toxic implication
PLOS ONE, 2017Co-Authors: Wenju Liu, Bo Zhou, Shoucai Fang, Hui Chen, Hao LiangAbstract:Growing evidence suggests that the pyrrolizidine alkaloids (PAs)-induced hepatotoxicity is mediated by multiple cell death/defence modalities. However, the detailed mechanisms are still lacking. In this study, the hepatotoxic effects of four PAs including three retronecine-type ones (senecionine, Seneciphylline and monocrotaline) and one otonecine-type (clivorine) on the proliferation of Huh-7.5 cells and the possible mechanisms were investigated. The results showed that all the PAs could inhibit cell proliferation and induce apoptosis in a concentration-dependent manner. Among them clivorine was the most significant one. In addition to its effect on apoptosis, clivorine treatment could promote autophagy in Huh-7.5 cells, as evidenced by the accumulation of autophagosomes, the enhancement of LC3B expression at the concentrations close to its IC0 value, and the increased conversion of LC3B-I to LC3B-II in the presence of lysosomal inhibitor (chloroquine) and decreased formation of green fluorescent protein (GFP)-LC3 positive puncta in the presence of autophagic sequestration inhibitor (3-methyladenine). Among the other tested PAs, senecionine and Seneciphylline also activated autophagy at the same concentrations used for clivorine but monocrotaline did not. Furthermore, our study demonstrated that suppression or enhancement of autophagy resulted in the remarkable enhancement or suppression of senecionine, Seneciphylline and clivorine-induced apoptosis at the concentration close to the IC10 for clivorine, respectively, indicating a protective role of autophagy against the PA-induced apoptosis at the low level of exposure. Collectively, our data suggest that PAs in different structures may exert different toxic disturbances on the liver cells. Apoptosis may be one of the most common models of the PA-induced cytotoxicity, while autophagy may be a structure-dependent defence model in the early stage of PA intoxication. Differential induction of apoptosis and autophagy probably depending on the concentration is essential for the cytotoxic potency of clivorine.
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Cytotoxic effects of senecionine, Seneciphylline, monocrotaline and clivorine increase with the increase of their concentrations in Huh-7.5 cells as determined by MTT.
2017Co-Authors: Wenju Liu, Bo Zhou, Shoucai Fang, Hui Chen, Hao Liang, Jun TangAbstract:Cells were incubated with indicated concentrations of senecionine (closed rectangle), Seneciphylline (closed diamond), monocrotaline (closed triangle) and clivorine (open circle) for 48 h, respectively. Cell viability was measured by MTT assay as described in Materials and Methods. Values are shown as mean ± SD (n = 3), of which the solvent control are at 0 μM; Δ, #P < 0.05, **P < 0.01, ***, ΔΔΔ, ###P < 0.001, comparing clivorine with senecionine (*), Seneciphylline (Δ), and monocrotaline (#), respectively, at the corresponding concentrations.
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The chemical structures of senecionine, Seneciphylline, monocrotaline and clivorine.
2017Co-Authors: Wenju Liu, Bo Zhou, Shoucai Fang, Hui Chen, Hao Liang, Jun TangAbstract:The chemical structures of senecionine, Seneciphylline, monocrotaline and clivorine.
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Transmission electron micrographs of autophagosome induced by senecionine, Seneciphylline, clivorine and monocrotaline in Huh-7.5 cells.
2017Co-Authors: Wenju Liu, Bo Zhou, Shoucai Fang, Hui Chen, Hao Liang, Jun TangAbstract:(a) The untreated Huh-7.5 cells (Control); (b) The senecionine -treated cells (6.25 μM), (c) The Seneciphylline-treated cells (6.25 μM), (d) The clivorine-treated cells (6.25 μM), (e) The monocrotaline-treated cells (6.25 μM). Huh-7.5 cells were cultured in the presence of senecionine, Seneciphylline, clivorine and monocrotaline for 24 h, respectively. As indicated by black arrows, the autophagic vacuoles with double-membrane structures were observed in cytoplasm. Asterisks indicate mature autophagosomes containing highly degraded contents. N, nucleus. Scale bar, 500 nm.
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Evaluation of senecionine, Seneciphylline, monocrotaline and clivorine-induced apoptosis of Huh-7.5 cells by flow cytometry analysis.
2017Co-Authors: Wenju Liu, Bo Zhou, Shoucai Fang, Hui Chen, Hao Liang, Jun TangAbstract:Cells were treated by four concentrations of senecionine, Seneciphylline, monocrotaline and clivorine for 24 h, respectively. Afterwards, the cells were collected and stained with Annexin V-kFluor488 and PI. The flow cytometry analysis was performed. The percentages of apoptotic cells at both the early and late periods were shown as Q1-LR and Q1-UR, respectively, in the form of scatter plots (a-1 to a-4). The results in the representative of three independent experiments were marked in each diagram and summarized in the histograms (b-1 to b-4). a-1 and b-1 show the results from senecionine-treated cells; a-2 and b-2, from Seneciphylline-treated cells; a-3 and b-3, from monocrotaline-treated cells; a-4 and b-4, from clivorine-treated cells. N0, F0, M0 and C0, negative controls; N6.25, N25, N100 and N400 represent senecionine-treated cells at the concentrations of 6.25 μM, 25 μM, 100 μM and 400 μM; F6.25, F25, F100 and F400 represent Seneciphylline-treated cells at the concentrations of 6.25 μM, 25 μM, 100 μM and 400 μM; M6.25, M25, M100 and M400 represent monocrotaline-treated cells at the concentrations of 6.25 μM, 25 μM, 100 μM and 400 μM; and C6.25, C25, C100, and C400 represent clivorine-treated cells at the concentrations of 6.25 μM, 25 μM, 100 μM and 400 μM, respectively. *P
