Z-十八碳-9-烯-丙磺酰胺对糖尿病认知功能障碍小鼠学习记忆功能的影响

探索新化合物Z-十八碳-9-烯-丙磺酰胺（N15）对糖尿病认知功能障碍（DACD）小鼠学习记忆功能的作用及机制。采用链脲佐菌素（STZ）连续小剂量腹腔注射以及高脂高糖加STZ诱导2型糖尿病小鼠模型,N15(50和100mg·kg-1·d-1)连续灌胃给药6周,于给药末期进行跳台、避暗以及Morris水迷宫测试以评价小鼠学习记忆功能;对海马内葡萄糖和乳酸水平进行测定;通过realtimePCR测定海马内突触成长相关蛋白-43（GAP-43）、突触素（SYN）、脑源性生长因子（BDNF）和神经营养因子-3（NT-3）m RNA的表达。结果表明,N15可显著改善模型小鼠学习记忆能力,降低海马内葡萄糖和乳酸含量,显著上调海马内GAP-43、SYN、BDNF和NT-3 m RNA表达水平。上述结果表明,新型化合物N15具有改善糖尿病认知功能障碍的作用,其机制可能与增加海马内突触生长相关因子和神经营养因子表达相关。福建省教育厅资助项目(JAT160583

Expression and Purification of Human Interferon-ω in Yeast Pichia pastoris

通信作者：[email protected][中文文摘]ω干扰素(IFN-ω)是同IFN-α功能相近但抗原性不同的一类干扰素分子,它具有同IFN-α、IFN-γ以及肿瘤坏死因子α相似的抑制病毒复制的作用.采用5L发酵罐发酵表达IFN-ω,发酵培养液经超滤膜浓缩、SP层析柱梯度分离,得到糖基化的IFN-ω,用基质辅助激光解析电离飞行时间质谱(MALDI TOF-TOF MS)和N端蛋白序列分析仪分析其分子质量和N端序列,用细胞病变抑制法测定活性.结果表明,诱导约48h时,目的蛋白表达量最高,通过SP梯度洗脱,可将糖基化和非糖基化的IFN-ω分离,质谱分析糖基化蛋白样品,质谱峰为糖基化蛋白特有的峰.纯化后糖基化的IFN-ω抗病毒活性为1.46×108 U/mL. [英文文摘]IFN-wis a kind of interferon molecules,which has similar function but different antigenicity to IFN－α.It can inhibit virus replication like IFN－α,IFN-γ and tumor necrosis factor alpha.5 L fermenter was used to ferment and interferon-ω was expressed.Then,the glycosylated interferon-ω was purified by a procedure stepped by ultrafiltration and gradient elution SP chromatography column.Its molecular weight and N amino acid sequence were analyzed by MALD I TOF-TOF MS and the N protein sequence analyzer,respectively.In addition,IFN-ω activity was 　determined by cell changes suppression method.The results showed that IFN-ω expression level reached to highest in induced 48h.The glycosylated and unglycosylated IFN－ω could be separated by SP gradient elution completely. The glycosylated IFN-ω showed a series of continuous peaks by mass spectrometry,which is the characteristic peak of glycosyl protein.The antiviral activity of 　the purified glycosylated IFN-ω was 1.46×108 u/／mL.国家自然科学基金项目(31071611);国家基础科学人才培养基金项目(J1030626