目的利用RP-HPLC法测定超声处理后五倍子中没食子酸,单宁酸,五没食子酰基葡萄糖的含量变化。方法干燥的五倍子粉碎,过80目筛,精密称取五倍子粉末各0.2 g分别置于三个烧杯中,分别加入p H为4的醋酸盐酸缓冲液40 m L,超声,取1 m L,过滤,进...目的利用RP-HPLC法测定超声处理后五倍子中没食子酸,单宁酸,五没食子酰基葡萄糖的含量变化。方法干燥的五倍子粉碎,过80目筛,精密称取五倍子粉末各0.2 g分别置于三个烧杯中,分别加入p H为4的醋酸盐酸缓冲液40 m L,超声,取1 m L,过滤,进样。结果五倍子中单宁酸及β-PGG在超声5分钟时含量达到最大值,随着超声时间的增加,单宁酸及β-PGG含量在5、20及40 min时呈下降趋势;五倍子中没食子酸在超声40 min时含量达到最大值,随着超声时间的增加,没食子酸的含量在5、20及40 min时呈上升趋势。结论建议从五倍子中提取单宁酸及β-PGG时不使用超声提取法,而五倍子中提取没食子酸时建议超声处理,且超声时间大于40 min,有利于没食子酸充分提取。本实验测定五倍子粉在不同超声时间没食子酸,单宁酸,β-PGG的含量变化,为优化五倍子中提取分离没食子酸,单宁酸,β-PGG等相关工艺提供一定实验基础。展开更多
This study examined the ability of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (β-PGG) to induce the expression of heme oxygenase-1 (HO-1) in the PC12 cells and its regulation in the PC12 cells.One week before treatment w...This study examined the ability of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (β-PGG) to induce the expression of heme oxygenase-1 (HO-1) in the PC12 cells and its regulation in the PC12 cells.One week before treatment with the drug,nerve growth factor (NGF) was added to the cultures at a final concentration of 50 ng/mL to induce neuronal differentiation.After drug treatment,HO-1 gene transcription was analyzed by reverse transcription polymerase chain reaction (RT-PCR).Expression of HO-1 and NF-E2-related factor2 (Nrf2) and activation of extracellular signal-regulated kinase (ERK) and Akt were detected by Western blotting.The viability of the PC12 cells treated with different medicines was examined by MTT assay.The oxidative stress in the PC12 cells was evaluated qualitatively and quantitatively by DCFH-DA.The results showed that β-PGG up-regulated HO-1 expression and this increased expression provided neuroprotection against MPP+-induced oxidative injury.Moreover,β-PGG induced Nrf2 nuclear translocation,which was found to be upstream of β-PGG-induced HO-1 expression,and the activation of ERK and Akt,a pathway that is involved in β-PGG-induced Nrf2 nuclear translocation,HO-1 expression and neuroprotection.In conclusion,β-PGG up-regulates HO-1 expression by stimulating Nrf2 nuclear translocation in an ERK-and Akt-dependent manner,and HO-1 expression by β-PGG may provide the PC12 cells with an acquired antioxidant defense capacity to survive the oxidative stress.展开更多
Objective： To understand the in vivo metabolic fate of 1,2,3,4,6-Penta-O-galloyl-β-d-glucose（PGG）naturally existed in many medicinal herbs and food plants such as Rhus chinensis,Paeonia suffruticosa,Paeonia lactif...Objective： To understand the in vivo metabolic fate of 1,2,3,4,6-Penta-O-galloyl-β-d-glucose（PGG）naturally existed in many medicinal herbs and food plants such as Rhus chinensis,Paeonia suffruticosa,Paeonia lactiflora and Mango.Methods： The metabolites of PGG in rat biofluids were characterized using high performance liquid chromatography combined with quadrupole time-of-flight tandem mass spectrometry（HPLC-QTOF-MS）.Results： Ten metabolites in urine,five metabolites in feces and two metabolites in plasma,were observed when the rats were administrated with a single intravenous injection of PGG（20 mg/kg）.Conclusion： PGG is firstly metabolized to gallic acid,then gallic acid undergoes sulfation,glucuronidation and methylation by rat liver.The determination of metabolites and the proposed metabolic pathway of PGG in vivo will be benefit to gain deeper insights into its pharmacological activities.展开更多
基金supported by National 11th Five-Year Plan Research Foundation of China (No.2006BAI01A14)
文摘This study examined the ability of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (β-PGG) to induce the expression of heme oxygenase-1 (HO-1) in the PC12 cells and its regulation in the PC12 cells.One week before treatment with the drug,nerve growth factor (NGF) was added to the cultures at a final concentration of 50 ng/mL to induce neuronal differentiation.After drug treatment,HO-1 gene transcription was analyzed by reverse transcription polymerase chain reaction (RT-PCR).Expression of HO-1 and NF-E2-related factor2 (Nrf2) and activation of extracellular signal-regulated kinase (ERK) and Akt were detected by Western blotting.The viability of the PC12 cells treated with different medicines was examined by MTT assay.The oxidative stress in the PC12 cells was evaluated qualitatively and quantitatively by DCFH-DA.The results showed that β-PGG up-regulated HO-1 expression and this increased expression provided neuroprotection against MPP+-induced oxidative injury.Moreover,β-PGG induced Nrf2 nuclear translocation,which was found to be upstream of β-PGG-induced HO-1 expression,and the activation of ERK and Akt,a pathway that is involved in β-PGG-induced Nrf2 nuclear translocation,HO-1 expression and neuroprotection.In conclusion,β-PGG up-regulates HO-1 expression by stimulating Nrf2 nuclear translocation in an ERK-and Akt-dependent manner,and HO-1 expression by β-PGG may provide the PC12 cells with an acquired antioxidant defense capacity to survive the oxidative stress.
基金the National Key Scientific Instrument and Development Project of China (2013YQ170525)the Program for Changjiang Scholars and Innovative Research Team in University (no.IRT15R55)+1 种基金National Natural Science Foundation of China (no.81603259)Natural Science Foundation of Shaanxi Province (2015JM2072)
文摘Objective： To understand the in vivo metabolic fate of 1,2,3,4,6-Penta-O-galloyl-β-d-glucose（PGG）naturally existed in many medicinal herbs and food plants such as Rhus chinensis,Paeonia suffruticosa,Paeonia lactiflora and Mango.Methods： The metabolites of PGG in rat biofluids were characterized using high performance liquid chromatography combined with quadrupole time-of-flight tandem mass spectrometry（HPLC-QTOF-MS）.Results： Ten metabolites in urine,five metabolites in feces and two metabolites in plasma,were observed when the rats were administrated with a single intravenous injection of PGG（20 mg/kg）.Conclusion： PGG is firstly metabolized to gallic acid,then gallic acid undergoes sulfation,glucuronidation and methylation by rat liver.The determination of metabolites and the proposed metabolic pathway of PGG in vivo will be benefit to gain deeper insights into its pharmacological activities.
