Diabetes have been shown to cause progressive neuronal injury with pain and numbness via advanced glycation end-products(AGEs)-induced neuronal cell apoptosis;however, the valuable drug targets for diabetic neuropathy...Diabetes have been shown to cause progressive neuronal injury with pain and numbness via advanced glycation end-products(AGEs)-induced neuronal cell apoptosis;however, the valuable drug targets for diabetic neuropathy have been poorly reported so far. In this study, we discovered a natural small-molecule schisandrol A(SolA) with significant protective effect against AGEs-induced neuronal cell apoptosis. ATP6V0D1, a major subunit of vacuolar-type ATPase(V-ATPase) in lysosome was identified as a crucial cellular target of SolA. Moreover, SolA allosterically mediated ATP6V0D1 conformation via targeting a unique cysteine 335 residue to activate V-ATPase-dependent lysosomal acidification.Interestingly, SolA-induced lysosome pH downregulation resulted in a mitochondrial-lysosomal crosstalk by selectively promoting mitochondrial BH3-only protein BIM degradation, thereby preserving mitochondrial homeostasis and neuronal cells survival. Collectively, our findings reveal ATP6V0D1 is a valuable pharmacological target for diabetes-associated neuronal injury via controlling lysosomal acidification, and also provide the first small-molecule template allosterically activating V-ATPase for preventing diabetic neuropathy.展开更多
目的建立反相高效液相色谱法同时测定五味子中6种木脂素含量的方法,并应用此方法检测南、北五味子及3种市售成药:参芪五味子片、安神补心片、安神补心胶囊中此6种木脂素的含量。方法采用SHIMADZU VP-ODS(4.6 mm×150 mm i.d,5μm)...目的建立反相高效液相色谱法同时测定五味子中6种木脂素含量的方法,并应用此方法检测南、北五味子及3种市售成药:参芪五味子片、安神补心片、安神补心胶囊中此6种木脂素的含量。方法采用SHIMADZU VP-ODS(4.6 mm×150 mm i.d,5μm)色谱柱,以乙腈-水为流动相,进行梯度洗脱,流速0.5mL/min,检测波长220 nm,柱温30℃。结果五味子醇甲、五味子醇乙、五味子酯甲、五味子甲素、五味子乙素和五味子丙素分离度良好,分别在0.243~2.430μg、0.087~0.870μg、0.060~0.600μg、0.093~0.930μg、0.246~2.460μg、0.063~0.630μg的进样量范围内有良好的线性关系,平均加样回收率分别为100.93%、100.59%、99.78%、100.43%、96.72%、102.33%,RSD值分别为2.14%、2.95%、2.26%、2.81%、1.88%、0.985%。结论本研究所建立的反相高效液相色谱法方法准确可靠、简单可行,可用于五味子中木脂素类成分的定量分析,为五味子药材及以五味子为组方药味的中成药的质量评价奠定基础。展开更多
基金supported by National Key Research and Development Program of China(Nos.2019YFC1708902 and 2019YFC1711000)National Natural Science Foundation of China(Nos.81973505,81773932 and 82104621).
文摘Diabetes have been shown to cause progressive neuronal injury with pain and numbness via advanced glycation end-products(AGEs)-induced neuronal cell apoptosis;however, the valuable drug targets for diabetic neuropathy have been poorly reported so far. In this study, we discovered a natural small-molecule schisandrol A(SolA) with significant protective effect against AGEs-induced neuronal cell apoptosis. ATP6V0D1, a major subunit of vacuolar-type ATPase(V-ATPase) in lysosome was identified as a crucial cellular target of SolA. Moreover, SolA allosterically mediated ATP6V0D1 conformation via targeting a unique cysteine 335 residue to activate V-ATPase-dependent lysosomal acidification.Interestingly, SolA-induced lysosome pH downregulation resulted in a mitochondrial-lysosomal crosstalk by selectively promoting mitochondrial BH3-only protein BIM degradation, thereby preserving mitochondrial homeostasis and neuronal cells survival. Collectively, our findings reveal ATP6V0D1 is a valuable pharmacological target for diabetes-associated neuronal injury via controlling lysosomal acidification, and also provide the first small-molecule template allosterically activating V-ATPase for preventing diabetic neuropathy.
