Aim To analyze the secondary structure and neurotrophic effect of a specific protein in sensory neurons. Methods Comparison of the proteins expressed in the rat spinal sensory neurons and motor neurons was made by t...Aim To analyze the secondary structure and neurotrophic effect of a specific protein in sensory neurons. Methods Comparison of the proteins expressed in the rat spinal sensory neurons and motor neurons was made by two dimensional electrophoresis. One specific protein in sensory neurons was isolated and purified by DEAE Sephacel ion exchange chromatography and high performance liquid chromatography. A primary analysis of its secondary structure by circular dichroism, and its neurotrophic effects were investigated using the model of dorsal root ganglia(DRG) cultured in vitro . Results The molecular weight and isoelectric point of the protein were 33 1 kDa and 5 52, respectively. Its circular dichroism showed that there were 20 8% α helix, 54 8% β sheet, 7 3% turn, and 17 1% random coil in its secondary structure. Biological experiments showed that the protein could promote the neurite outgrowth of DRG. Conclusion A specific protein in spinal sensory tissue with molecular weight of 33 1 kDa has been purified. There is mainly β sheet in the secondary structure of the protein. And the protein has neurotrophic effects in the model of DRG.展开更多
A green, rapid and precise sample pretreatment technique, IL-based UAE（ionic liquid-based ultrasonic- assisted extraction）, was coupled with high-performance liquid chromatographic separation to identify the main ef...A green, rapid and precise sample pretreatment technique, IL-based UAE（ionic liquid-based ultrasonic- assisted extraction）, was coupled with high-performance liquid chromatographic separation to identify the main effective components in Schisandra sphenanthera（S, sphenanthera） and Schisandra chinensis（S, ehinensis） including schisantherin A, schisandrin A, and deoxyschizandrin. Four different types of ionic liquids have been investigated, finally [C6MIM][BF4] was used as the extraction solvent. A powder form of S. sphenanthera and S. chinensis was mixed with the [C6MIM][BF4] to produce a suspension. This suspension was ultrasonically extracted in a water bath at room temperature. Several of the process parameters were optimized, including the type of ionic liquid used and its volume, the sample amount, the size of the sample particle, the extraction time, etc. HPLC calibration curves were estab-lished for all the analytes and proved to be linear（r〉0.9999）. The lowest detection level for schisandrin A was 0.12μg/mL, for schisantherin A was 0.08 μg/mL, and for deoxyschizandfin was 0.10μg/mL. The recoveries of the target compounds were from 74.19% to 109.33%. The standard deviations for detection were generally no more than 6.31%. In contrast to conventional extraction methods, the IL-based UAE did not involve volatile organic volatile solvents, and the analysis time, required sample and solvent vohtrnes were also lower than those of the conventional techniques.展开更多
文摘Aim To analyze the secondary structure and neurotrophic effect of a specific protein in sensory neurons. Methods Comparison of the proteins expressed in the rat spinal sensory neurons and motor neurons was made by two dimensional electrophoresis. One specific protein in sensory neurons was isolated and purified by DEAE Sephacel ion exchange chromatography and high performance liquid chromatography. A primary analysis of its secondary structure by circular dichroism, and its neurotrophic effects were investigated using the model of dorsal root ganglia(DRG) cultured in vitro . Results The molecular weight and isoelectric point of the protein were 33 1 kDa and 5 52, respectively. Its circular dichroism showed that there were 20 8% α helix, 54 8% β sheet, 7 3% turn, and 17 1% random coil in its secondary structure. Biological experiments showed that the protein could promote the neurite outgrowth of DRG. Conclusion A specific protein in spinal sensory tissue with molecular weight of 33 1 kDa has been purified. There is mainly β sheet in the secondary structure of the protein. And the protein has neurotrophic effects in the model of DRG.
基金Supported by the National Natural Science Foundation of China（Nos. 81671220, 81502516） and the Jilin Provincial School Joint Construction Special Project, China（No.SXGJQY2017-13）.
文摘A green, rapid and precise sample pretreatment technique, IL-based UAE（ionic liquid-based ultrasonic- assisted extraction）, was coupled with high-performance liquid chromatographic separation to identify the main effective components in Schisandra sphenanthera（S, sphenanthera） and Schisandra chinensis（S, ehinensis） including schisantherin A, schisandrin A, and deoxyschizandrin. Four different types of ionic liquids have been investigated, finally [C6MIM][BF4] was used as the extraction solvent. A powder form of S. sphenanthera and S. chinensis was mixed with the [C6MIM][BF4] to produce a suspension. This suspension was ultrasonically extracted in a water bath at room temperature. Several of the process parameters were optimized, including the type of ionic liquid used and its volume, the sample amount, the size of the sample particle, the extraction time, etc. HPLC calibration curves were estab-lished for all the analytes and proved to be linear（r〉0.9999）. The lowest detection level for schisandrin A was 0.12μg/mL, for schisantherin A was 0.08 μg/mL, and for deoxyschizandfin was 0.10μg/mL. The recoveries of the target compounds were from 74.19% to 109.33%. The standard deviations for detection were generally no more than 6.31%. In contrast to conventional extraction methods, the IL-based UAE did not involve volatile organic volatile solvents, and the analysis time, required sample and solvent vohtrnes were also lower than those of the conventional techniques.
