Precision-cut liver slice has been successfully used to study the mechanism of drug-induced hepatotoxicity, the prediction of liver toxicity, the discovery of early hepatic toxicity biomarker and the metabolism of dru...Precision-cut liver slice has been successfully used to study the mechanism of drug-induced hepatotoxicity, the prediction of liver toxicity, the discovery of early hepatic toxicity biomarker and the metabolism of drug in liver. We detected the expression of CYP3A4, CYP2B1 + CYP2B2 and CYP2E1 in precision-cut liver slice after co-cultured with monocrotaline or Tussilago farfara alkaloids to investigate the hepatotoxicity mechanism of those drugs. After co-culturing with monocrotaline or Tussilago farfara alkaloids for 6 hours, the expression of CYP3A4 in the microsome of precision-cut liver slices was detected by Western blot, and the expressions of CYP2B1 + CYP2B2 and CYP2E1 were detected by immunofluorescence. The results showed that monocrotaline induced the expression of CYP3A4 and CYP2B1 + CYP2B2, and Tussilago farfara alkaloids obviously up-regulated the expression of CYP2E1 and CYP3A4. Thus, we conclude that the up-regulation of CYP3A4, CYP2B1 + CYP2B2 and CYP2E1 may be one of the toxic mechanisms of liver injury of those drugs.展开更多
Objective： To investigate the effects of the flower buds extract of Tussilago farfara Linné（Farfarae Flos; FF） on focal cerebral ischemia through regulation of inflammatory responses in activated microglia. Me...Objective： To investigate the effects of the flower buds extract of Tussilago farfara Linné（Farfarae Flos; FF） on focal cerebral ischemia through regulation of inflammatory responses in activated microglia. Methods： Brain ischemia was induced in Sprague-Dawley rats by a transient middle cerebral artery occlusion（t MCAO） for 90 min and reperfusion for 24 h. Twenty rats were randomly divided into 4 groups（n=5 per group）： normal, t MCAO-induced ischemic control, t MCAO plus FF extract 300 mg/kg-treated, and t MCAO plus MK-801 1 mg/kg-treated as reference drug. FF extract（300 mg/kg, p.o.） or MK-801（1 mg/kg, i.p.） was administered after reperfusion. Brain infarction was measured by 2,3,5,-triphenyltetrazolium chloride staining. Neuronal damage was observed by haematoxylin eosin, Nissl staining and immunohistochemistry using anti-neuronal nuclei（Neu N）, anti-glial fibril ary acidic protein（GFAP）, and antiCD11 b/c（OX42） antibodies in ischemic brain. The expressions of inducible nitric oxide synthase（i NOS）, tumor necrosis factor（TNF-α）, and hypoxia-inducible factor-1 a（HIF-1α） were determined by Western blot. BV2 microglial cel s were treated with FF extract or its main bioactive compound, tussilagone with or without lipopolysaccharide（LPS）. Nitric oxide（NO） production was measured in culture medium by Griess assay. The expressions of i NOS, COX-2 and pro-inflammatory cytokines mRNA were analyzed by reverse transcription-polymerase chain reaction. The expression of i NOS, and COX-2 proteins, the phosphorylation of ERK1/2, JNK, and p38 MAPK and the nuclear expression of NF-κB p65 in BV2 cells were determined by Western blot. Results： FF extract significantly decreased brain infarctions in ischemic rats（P〈0.01）. The neuronal death and the microglia/astrocytes activation in ischemic brains were inhibited by FF extract. FF extract also suppressed i NOS, TNF-α, and HIF-1α expression in ischemic brains. FF extract（0.2 and 0.5 mg/m L, P〈0.01） and tussilagone 20 and 50 μmol/L, P〈0.01） significantly decreased LPS-induced NO production in BV2 microglia through downregulation of i NOS m RNA and protein expression. FF extract and tussilagone significantly inhibited LPS-induced expression of TNF-α, IL-1β, and IL-6 m RNA, and also suppressed the phosphorylation of ERK1/2, JNK and p38 MAPK and the nuclear expression of NF-κB in a dosedependent manner. Conclusion： FF extract has a neuroprotective effect in ischemic stroke by the decrease of brain infarction, and the inhibition of neuronal death and microglial activation-mediated inflammatory responses.展开更多
Chemical fractionation of the n-BuOH partition,which was generated from the EtOH extract of the flower buds of Tussilago farfara,afforded a series of polar constituents including four new sesquiterpenoids(1-4),one new...Chemical fractionation of the n-BuOH partition,which was generated from the EtOH extract of the flower buds of Tussilago farfara,afforded a series of polar constituents including four new sesquiterpenoids(1-4),one new sesquiterpenoid glucoside(5)and one known analogue(6)of the eudesmane type,as well as five known quinic acid derivatives(7-11).Structures of the new compounds were unambiguously characterized by detailed spectroscopic analyses,with their absolute configurations being established by A-ray crystallography,electronic circular dichroism(ECD)calculation and induced ECD experiments.The inhibitory effect of all the isolates against LPS-induced NO production in murine RAW264.7 macrophages was evaluated,with isochlorogenic acid A(7)showing significant inhibitory activity.展开更多
文摘Precision-cut liver slice has been successfully used to study the mechanism of drug-induced hepatotoxicity, the prediction of liver toxicity, the discovery of early hepatic toxicity biomarker and the metabolism of drug in liver. We detected the expression of CYP3A4, CYP2B1 + CYP2B2 and CYP2E1 in precision-cut liver slice after co-cultured with monocrotaline or Tussilago farfara alkaloids to investigate the hepatotoxicity mechanism of those drugs. After co-culturing with monocrotaline or Tussilago farfara alkaloids for 6 hours, the expression of CYP3A4 in the microsome of precision-cut liver slices was detected by Western blot, and the expressions of CYP2B1 + CYP2B2 and CYP2E1 were detected by immunofluorescence. The results showed that monocrotaline induced the expression of CYP3A4 and CYP2B1 + CYP2B2, and Tussilago farfara alkaloids obviously up-regulated the expression of CYP2E1 and CYP3A4. Thus, we conclude that the up-regulation of CYP3A4, CYP2B1 + CYP2B2 and CYP2E1 may be one of the toxic mechanisms of liver injury of those drugs.
基金Supported by the grant from Ministry of Food and Drug Safety in 2014(No.12172MFDS989)Republic of Korea
文摘Objective： To investigate the effects of the flower buds extract of Tussilago farfara Linné（Farfarae Flos; FF） on focal cerebral ischemia through regulation of inflammatory responses in activated microglia. Methods： Brain ischemia was induced in Sprague-Dawley rats by a transient middle cerebral artery occlusion（t MCAO） for 90 min and reperfusion for 24 h. Twenty rats were randomly divided into 4 groups（n=5 per group）： normal, t MCAO-induced ischemic control, t MCAO plus FF extract 300 mg/kg-treated, and t MCAO plus MK-801 1 mg/kg-treated as reference drug. FF extract（300 mg/kg, p.o.） or MK-801（1 mg/kg, i.p.） was administered after reperfusion. Brain infarction was measured by 2,3,5,-triphenyltetrazolium chloride staining. Neuronal damage was observed by haematoxylin eosin, Nissl staining and immunohistochemistry using anti-neuronal nuclei（Neu N）, anti-glial fibril ary acidic protein（GFAP）, and antiCD11 b/c（OX42） antibodies in ischemic brain. The expressions of inducible nitric oxide synthase（i NOS）, tumor necrosis factor（TNF-α）, and hypoxia-inducible factor-1 a（HIF-1α） were determined by Western blot. BV2 microglial cel s were treated with FF extract or its main bioactive compound, tussilagone with or without lipopolysaccharide（LPS）. Nitric oxide（NO） production was measured in culture medium by Griess assay. The expressions of i NOS, COX-2 and pro-inflammatory cytokines mRNA were analyzed by reverse transcription-polymerase chain reaction. The expression of i NOS, and COX-2 proteins, the phosphorylation of ERK1/2, JNK, and p38 MAPK and the nuclear expression of NF-κB p65 in BV2 cells were determined by Western blot. Results： FF extract significantly decreased brain infarctions in ischemic rats（P〈0.01）. The neuronal death and the microglia/astrocytes activation in ischemic brains were inhibited by FF extract. FF extract also suppressed i NOS, TNF-α, and HIF-1α expression in ischemic brains. FF extract（0.2 and 0.5 mg/m L, P〈0.01） and tussilagone 20 and 50 μmol/L, P〈0.01） significantly decreased LPS-induced NO production in BV2 microglia through downregulation of i NOS m RNA and protein expression. FF extract and tussilagone significantly inhibited LPS-induced expression of TNF-α, IL-1β, and IL-6 m RNA, and also suppressed the phosphorylation of ERK1/2, JNK and p38 MAPK and the nuclear expression of NF-κB in a dosedependent manner. Conclusion： FF extract has a neuroprotective effect in ischemic stroke by the decrease of brain infarction, and the inhibition of neuronal death and microglial activation-mediated inflammatory responses.
基金This work was supported by the Natural Science Foundation of Shandong Province(No.JQ201721)the Young Taishan Scholars Program(No.tsqn20161037)Innovation Team Project of Jinan Science&Technology Bureau(No.2018GXRC003).
文摘Chemical fractionation of the n-BuOH partition,which was generated from the EtOH extract of the flower buds of Tussilago farfara,afforded a series of polar constituents including four new sesquiterpenoids(1-4),one new sesquiterpenoid glucoside(5)and one known analogue(6)of the eudesmane type,as well as five known quinic acid derivatives(7-11).Structures of the new compounds were unambiguously characterized by detailed spectroscopic analyses,with their absolute configurations being established by A-ray crystallography,electronic circular dichroism(ECD)calculation and induced ECD experiments.The inhibitory effect of all the isolates against LPS-induced NO production in murine RAW264.7 macrophages was evaluated,with isochlorogenic acid A(7)showing significant inhibitory activity.
