黑水缬草提取物对阿尔茨海默病模型小鼠治疗作用的代谢组学研究

Metabonomics study on the effect of Valeriana amurensis extractive for model mice with Alzheimer disease

目的运用超高效液相色谱-质谱联用技术(UPLC-MS)探讨黑水缬草提取物对阿尔茨海默病(AD)的治疗机制。方法将雄性昆明小鼠分为3组,分别为对照组(2%吐温80)、AD模型组(2%吐温80)和治疗组(95%黑水缬草粗提物2 g/kg),灌胃给药30 d。运用Morris水迷宫实验和UPLC-MS技术对小鼠进行研究。结果 Morris水迷宫定位航行实验显示,与对照组比较,AD模型组小鼠逃避潜伏期明显延长(P<0.05),治疗组小鼠逃避潜伏期明显缩短(P<0.05)。空间探索实验显示,与对照组比较,AD模型组小鼠单位时间目标象限停留时间短,穿台次数减少(P<0.05),治疗组小鼠单位时间内目标象限停留时间长,穿台次数增加(P<0.05)。通过UPLC-MS/MS技术,确定了10种AD的血浆生物标志物,分别为组氨酸、溶血磷脂酰胆碱C16∶0、溶血磷脂酰胆碱C18∶2、溶血磷脂酰胆碱C18∶1、溶血磷脂酰胆碱C20∶4、溶血磷脂酰胆碱C22∶6、十六碳鞘磷脂、二氢神经鞘氨醇、4-羟双氢(神经)鞘氨醇C18、4-羟双氢(神经)鞘氨醇C20,与对照组比较,AD模型组小鼠血浆中10种标志物的含量均明显降低,差异有统计学意义(P<0.05或P<0.01)。治疗组小鼠血浆中10种标志物的含量均明显上升(P<0.05或P<0.01)。结论黑水缬草提取物可提高AD小鼠的认知功能,其作用机制与氨基酸、磷脂和鞘脂代谢等相关代谢通路有关。

Objective To investigate the therapeutic mechanism of Valeriana amurensis extractive for Alzheimer dis- ease （AD） by ultra-performance liquid chromatography-mass spectrometry （UPLC-MS）. Methods KM male mice were divided into three groups： control group （2% Tween-80 solvent）, AD model group （2% Tween-80 solvent）, treatment group （95% EtOH extract of Valeriana amurensis 2 g/kg）, all mice were given drugs by gavage for 30 days. Mortis wa- ter maze and UPLC-MS/MS technique were conducted in the experiment. Results In the place navigation test, mice in the AD group took much more time to reach the platform on the last three training days compared to the control group （P 〈 0.05）. The treatment group reduced escape latency compared to the AD group on last 4 days （P 〈 0.05）. In the spatial exploration test, the total time spent in the target quadrant and the number of mice that exactly crossed the pre- vious position of the platform in AD model group were clearly shorter and lower than those of the control group （P 〈 0.05）. However, mice in the treatment group showed longer time spent in the target quadrant and higher frequencies of platform crossings compared to the AD group （P 〈 0.05）. By means of UPLC-MS, a total of ten kinds of analytes were identified, which were L-Histidine, C18 PHS, Hexadecasphinganine, LPC C22：6, LPC C18：2, LPC C16：0, LPC C20：4, LPC C18：1, dihydrosphingosine and C20 PHS. When compared to control group, the levels of ten kinds of markers in AD model group were Significantly reduced （P 〈 0.05 or P 〈 0.01）. But the contents of ten kinds of markers in mice plasma in the treatment group were obviously increased （P 〈 0.05 or P 〈 0.01）. Conclusion Valeriana amurensis extract can improve the cognitive tunetion of AD mice, its mechanism is closely related to amino acids, phospho- lipids and sheath lipid metabolism as well as related metabolic pathways.