Objective:To optimize the extraction process of total flavones in Trichosanthis Fructus(composed of Trichosanthis pericarpium and Trichosanthis semen in certain proportion).Methods:The effects of the mixture ratio of ...Objective:To optimize the extraction process of total flavones in Trichosanthis Fructus(composed of Trichosanthis pericarpium and Trichosanthis semen in certain proportion).Methods:The effects of the mixture ratio of Trichosanthis pericarpium and Trichosanthis semen,ethanol concentration,ultrasonic extraction time and extraction temperature on the extraction rate of total flavonoids in Trichosanthis Fructus were investigated.The extraction process of total flavonoids in Trichosanthis Fructus was optimized by Box-Behnken response surface method combined with differential spectrophotometry.Results:The optimum extraction conditions of total flavonoids in Trichosanthis Fructus were as follows:The mixture ratio of Trichosanthis pericarpium and Trichosanthis semen was 4:6,the ethanol concentration was 70%,the ultrasonic extraction time was 60min and the extraction temperature was 40℃.Conclusion:Box-Behnken response surface method combined with differential spectrophotometry can optimize the extraction process of total flavonoids from Trichosanthis Fructus,which can provide reference for the extraction and application of total flavonoids in Trichosanthis Fructus.展开更多
目的观察瓜蒌皮提取物对AngⅡ诱导血管平滑肌细胞增殖的影响。方法采用MTS法检测瓜蒌皮提取物(10、20、30 mg·L-1)对血管紧张素Ⅱ(10-7mol·L-1)所致血管平滑肌细胞增殖活性的影响;Real time RTPCR法检测血管平滑肌细胞中内皮...目的观察瓜蒌皮提取物对AngⅡ诱导血管平滑肌细胞增殖的影响。方法采用MTS法检测瓜蒌皮提取物(10、20、30 mg·L-1)对血管紧张素Ⅱ(10-7mol·L-1)所致血管平滑肌细胞增殖活性的影响;Real time RTPCR法检测血管平滑肌细胞中内皮型一氧化氮合酶、原癌基因c-fos。结果 MTS检测结果示,与对照组相比,AngⅡ组A490值明显升高(P<0.05),与AngⅡ组相比,AngⅡ+EPT组A490值均明显下降(P<0.05)。Real-time RT-PCR测定结果示,AngⅡ组e NOS mRNA的表达低于对照组,c-fos mRNA的表达高于对照组,差异均有统计学意义(P<0.05);加入EPT后,e NOS mRNA的表达高于AngⅡ组,c-fos mRNA的表达低于AngⅡ组,差异均有统计学意义(P<0.05)。结论瓜蒌皮提取物抑制血管紧张素Ⅱ所致的细胞增殖,其作用机制可能与降低c-fos mRNA高表达,升高一氧化氮合酶mRNA表达有关。展开更多
目的:研究磷脂酰肌醇3-激酶/蛋白激酶B/一氧化氮(PI3K/Akt/NO)信号通路在瓜蒌皮提取物保护心肌细胞缺氧/复氧损伤中的作用。方法:以2.5 mmol·L-1Na2S2O4诱导心肌细胞建立缺氧/复氧损伤模型,实验分为正常组、模型组、瓜蒌皮提取物...目的:研究磷脂酰肌醇3-激酶/蛋白激酶B/一氧化氮(PI3K/Akt/NO)信号通路在瓜蒌皮提取物保护心肌细胞缺氧/复氧损伤中的作用。方法:以2.5 mmol·L-1Na2S2O4诱导心肌细胞建立缺氧/复氧损伤模型,实验分为正常组、模型组、瓜蒌皮提取物组及抑制剂组。采用噻唑蓝(MTT)比色法检测细胞活力;酶联免疫吸附测定(ELISA)检测心肌细胞中一氧化氮(NO)的释放量及内皮型一氧化氮合酶(e NOS),诱导型一氧化氮合酶(i NOS)的活性水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测细胞中Akt,e NOS,i NOS mRNA表达水平;蛋白免疫印迹法(Western blot)测定Akt,磷酸化Akt(p-Akt),e NOS,i NOS蛋白表达水平。结果:与正常组比较,模型组心肌细胞活力明显下降(P<0.01),心肌细胞NO释放量减少(P<0.01),心肌细胞中p-Akt,Akt及e NOS mRNA和蛋白表达显著降低(P<0.01),而i NOS mRNA和蛋白表达显著升高(P<0.01);与模型组比较,瓜蒌皮提取物预处理24 h后能改善心肌细胞形态,明显提高心肌细胞活力(P<0.01)。瓜蒌皮提取物能促进p-Akt,Akt及e NOS mRNA和蛋白表达(P<0.01),抑制i NOS mRNA和蛋白的表达(P<0.01),增加心肌细胞NO的释放量(P<0.01)。PI3K抑制剂LY294002与瓜蒌皮提取物联合处理后,逆转了瓜蒌皮提取物对心肌细胞的上述作用,导致细胞存活率下降。结论:瓜蒌皮提取物可能通过激活PI3K/Akt信号通路上调e NOS,下调i NOS的表达,增加内源性NO的基础含量水平,发挥抗凋亡作用,从而改善缺氧/复氧损伤诱导的心肌细胞损伤。展开更多
基金Anhui Universities Provincial Key Project of Natural Science Research(No.KJ2016SD60,KJ2015ZD41)。
文摘Objective:To optimize the extraction process of total flavones in Trichosanthis Fructus(composed of Trichosanthis pericarpium and Trichosanthis semen in certain proportion).Methods:The effects of the mixture ratio of Trichosanthis pericarpium and Trichosanthis semen,ethanol concentration,ultrasonic extraction time and extraction temperature on the extraction rate of total flavonoids in Trichosanthis Fructus were investigated.The extraction process of total flavonoids in Trichosanthis Fructus was optimized by Box-Behnken response surface method combined with differential spectrophotometry.Results:The optimum extraction conditions of total flavonoids in Trichosanthis Fructus were as follows:The mixture ratio of Trichosanthis pericarpium and Trichosanthis semen was 4:6,the ethanol concentration was 70%,the ultrasonic extraction time was 60min and the extraction temperature was 40℃.Conclusion:Box-Behnken response surface method combined with differential spectrophotometry can optimize the extraction process of total flavonoids from Trichosanthis Fructus,which can provide reference for the extraction and application of total flavonoids in Trichosanthis Fructus.
文摘目的观察瓜蒌皮提取物对AngⅡ诱导血管平滑肌细胞增殖的影响。方法采用MTS法检测瓜蒌皮提取物(10、20、30 mg·L-1)对血管紧张素Ⅱ(10-7mol·L-1)所致血管平滑肌细胞增殖活性的影响;Real time RTPCR法检测血管平滑肌细胞中内皮型一氧化氮合酶、原癌基因c-fos。结果 MTS检测结果示,与对照组相比,AngⅡ组A490值明显升高(P<0.05),与AngⅡ组相比,AngⅡ+EPT组A490值均明显下降(P<0.05)。Real-time RT-PCR测定结果示,AngⅡ组e NOS mRNA的表达低于对照组,c-fos mRNA的表达高于对照组,差异均有统计学意义(P<0.05);加入EPT后,e NOS mRNA的表达高于AngⅡ组,c-fos mRNA的表达低于AngⅡ组,差异均有统计学意义(P<0.05)。结论瓜蒌皮提取物抑制血管紧张素Ⅱ所致的细胞增殖,其作用机制可能与降低c-fos mRNA高表达,升高一氧化氮合酶mRNA表达有关。
文摘目的:研究磷脂酰肌醇3-激酶/蛋白激酶B/一氧化氮(PI3K/Akt/NO)信号通路在瓜蒌皮提取物保护心肌细胞缺氧/复氧损伤中的作用。方法:以2.5 mmol·L-1Na2S2O4诱导心肌细胞建立缺氧/复氧损伤模型,实验分为正常组、模型组、瓜蒌皮提取物组及抑制剂组。采用噻唑蓝(MTT)比色法检测细胞活力;酶联免疫吸附测定(ELISA)检测心肌细胞中一氧化氮(NO)的释放量及内皮型一氧化氮合酶(e NOS),诱导型一氧化氮合酶(i NOS)的活性水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测细胞中Akt,e NOS,i NOS mRNA表达水平;蛋白免疫印迹法(Western blot)测定Akt,磷酸化Akt(p-Akt),e NOS,i NOS蛋白表达水平。结果:与正常组比较,模型组心肌细胞活力明显下降(P<0.01),心肌细胞NO释放量减少(P<0.01),心肌细胞中p-Akt,Akt及e NOS mRNA和蛋白表达显著降低(P<0.01),而i NOS mRNA和蛋白表达显著升高(P<0.01);与模型组比较,瓜蒌皮提取物预处理24 h后能改善心肌细胞形态,明显提高心肌细胞活力(P<0.01)。瓜蒌皮提取物能促进p-Akt,Akt及e NOS mRNA和蛋白表达(P<0.01),抑制i NOS mRNA和蛋白的表达(P<0.01),增加心肌细胞NO的释放量(P<0.01)。PI3K抑制剂LY294002与瓜蒌皮提取物联合处理后,逆转了瓜蒌皮提取物对心肌细胞的上述作用,导致细胞存活率下降。结论:瓜蒌皮提取物可能通过激活PI3K/Akt信号通路上调e NOS,下调i NOS的表达,增加内源性NO的基础含量水平,发挥抗凋亡作用,从而改善缺氧/复氧损伤诱导的心肌细胞损伤。
