OBJECTIVE Mu-Xiang-You-Fang(MXYF)is a classic prescription of Hui medicine,composed of five herbs,which has been used to treat ischemic stroke for many years.However,the potential pharmacological mecha⁃nisms of MXYF r...OBJECTIVE Mu-Xiang-You-Fang(MXYF)is a classic prescription of Hui medicine,composed of five herbs,which has been used to treat ischemic stroke for many years.However,the potential pharmacological mecha⁃nisms of MXYF remain unclear.The present research is to investigate the neuroprotective effect of MXYF and its role in modulating autophagy via AMPK/mTOR signaling pathway in the PC12 oxygen-glucose deprivation and reperfusion(OGD/R)injury model.METHODS MXYF was extracted by supercritical CO2 fluid extraction apparatus.PC12 OGD/R injury model was established by oxygen-glucose deprivation for 2 h and reperfusion for 24 h.The effects of MXYF on the viability and cytotoxicity of PC12 cells were determined through cell counting kit(CCK-8)assay.Colorimetric method was performed to determine the LDH leakage rate.The calcium concentration was determined by chemical fluorescence method and the mitochondrial membrane potential was determined through flow cytometry.Monodansylcadaverine(MDC)staining was conducted to detect autophagosome formation.The expression of LC3,Beclin1,p62,p-AMPK,ULK1,p-mTOR and p-p70s6k proteins were determined by immunofluorescence and Western blotting analyses.RESULTS MXYF(1,2 and 4 mg·L^-1)could significantly increase the cell viability and mitochondrial membrane potential,while decreased the release of lactate dehydrogenase(LDH)and calcium concentration in PC12 cells.Mechanistic studies showed that MXYF reduced the LC3-II/LC3-I ratio and inhibited the expression of beclin1,p-AMPK and ULK1.In comparison,the expres⁃sion of p-mTOR,p-p70s6k and p62 were significantly enhanced.CONCLUSION MXYF inhibits autophagy after OGD/Rinduced PC12 cell injury through AMPK-mTOR pathway,thus MXYF might have therapeutic potential for treating the ischemic stroke.展开更多
近年来随着学术交流的不断深入,美国芝加哥菲尔德自然历史博物馆(Field Museum of Natural History)收藏的中国碑帖拓本日益为人们所关注,《宋拓赵侍郎不流本游相〈兰亭〉》就是其中之一。该本为游相《兰亭》"己之二"(第五十...近年来随着学术交流的不断深入,美国芝加哥菲尔德自然历史博物馆(Field Museum of Natural History)收藏的中国碑帖拓本日益为人们所关注,《宋拓赵侍郎不流本游相〈兰亭〉》就是其中之一。该本为游相《兰亭》"己之二"(第五十二种),翻刻于一"九字已损"、"五字未损"的定武古本。由于卷改册装,其上"赵氏孟林"骑缝印被一分为二,十分罕见。在目前关于"赵氏孟林"的文献记载尚不充分、明确的情况下,可以从已知的游相《兰亭》拓本装潢钤印定式本身入手,对其进行分析。检览统计若干游相《兰亭》相关信息,笔者认为,赵孟林是明初晋藩朱棡装潢人的可能性较大。作为游相《兰亭》标志要素之一的蓝纸隔水,则是经朱氏收藏后重新进行装裱的一个显著特征。而同一取材、或有或无的蓝纸小签上所标注的"天干编次帖本名称",亦是游相《兰亭》标志要素之一,或许正是出自赵孟林之手,当然也不排除朱棡亲笔的可能。展开更多
基金National Natural Science Foundation of China(8166070081260679)Ningxia College FirstClass Discipline Construction Project(Chinese Medicine)Funded Project(NXYLXK2017A06)
文摘OBJECTIVE Mu-Xiang-You-Fang(MXYF)is a classic prescription of Hui medicine,composed of five herbs,which has been used to treat ischemic stroke for many years.However,the potential pharmacological mecha⁃nisms of MXYF remain unclear.The present research is to investigate the neuroprotective effect of MXYF and its role in modulating autophagy via AMPK/mTOR signaling pathway in the PC12 oxygen-glucose deprivation and reperfusion(OGD/R)injury model.METHODS MXYF was extracted by supercritical CO2 fluid extraction apparatus.PC12 OGD/R injury model was established by oxygen-glucose deprivation for 2 h and reperfusion for 24 h.The effects of MXYF on the viability and cytotoxicity of PC12 cells were determined through cell counting kit(CCK-8)assay.Colorimetric method was performed to determine the LDH leakage rate.The calcium concentration was determined by chemical fluorescence method and the mitochondrial membrane potential was determined through flow cytometry.Monodansylcadaverine(MDC)staining was conducted to detect autophagosome formation.The expression of LC3,Beclin1,p62,p-AMPK,ULK1,p-mTOR and p-p70s6k proteins were determined by immunofluorescence and Western blotting analyses.RESULTS MXYF(1,2 and 4 mg·L^-1)could significantly increase the cell viability and mitochondrial membrane potential,while decreased the release of lactate dehydrogenase(LDH)and calcium concentration in PC12 cells.Mechanistic studies showed that MXYF reduced the LC3-II/LC3-I ratio and inhibited the expression of beclin1,p-AMPK and ULK1.In comparison,the expres⁃sion of p-mTOR,p-p70s6k and p62 were significantly enhanced.CONCLUSION MXYF inhibits autophagy after OGD/Rinduced PC12 cell injury through AMPK-mTOR pathway,thus MXYF might have therapeutic potential for treating the ischemic stroke.
文摘近年来随着学术交流的不断深入,美国芝加哥菲尔德自然历史博物馆(Field Museum of Natural History)收藏的中国碑帖拓本日益为人们所关注,《宋拓赵侍郎不流本游相〈兰亭〉》就是其中之一。该本为游相《兰亭》"己之二"(第五十二种),翻刻于一"九字已损"、"五字未损"的定武古本。由于卷改册装,其上"赵氏孟林"骑缝印被一分为二,十分罕见。在目前关于"赵氏孟林"的文献记载尚不充分、明确的情况下,可以从已知的游相《兰亭》拓本装潢钤印定式本身入手,对其进行分析。检览统计若干游相《兰亭》相关信息,笔者认为,赵孟林是明初晋藩朱棡装潢人的可能性较大。作为游相《兰亭》标志要素之一的蓝纸隔水,则是经朱氏收藏后重新进行装裱的一个显著特征。而同一取材、或有或无的蓝纸小签上所标注的"天干编次帖本名称",亦是游相《兰亭》标志要素之一,或许正是出自赵孟林之手,当然也不排除朱棡亲笔的可能。
