黑水缬草提取物对Aβ_(25-35)致阿尔茨海默病模型小鼠生物标记物调控作用的研究

Study on the biomarkers regulation mechanism of Valeriana amurensis extractive on Aβ_(25-35) induced Alzheimer's disease mouse model

目的:运用超高效液相色谱质谱(UPLC-MS)技术研究黑水缬草提取物对Aβ_(25-35)诱导所致阿尔茨海默病(AD)模型小鼠的干预机制,为AD治疗提供新的思路。方法:将实验小鼠随机分为3组:对照组、AD模型组和治疗组,运用免疫组化实验观察AD小鼠脑部组织病理学改变,运用UPLC-MS技术研究黑水缬草提取物对AD模型小鼠生物标记物的调控作用。结果:对照组小鼠海马区细胞排列整齐且形态正常。AD模型组小鼠海马区可见到大量淀粉样斑块。治疗组小鼠海马区淀粉样斑块数量减少,细胞趋于正常。通过对实验小鼠血浆代谢组学进行分析,确定了色氨酸、组氨酸、溶血磷脂酰胆碱(LPC)C22∶6、LPC C18∶2、LPC C16∶0(m/z 496.2)、LPC C16∶0(m/z 991.6)、二氢神经鞘氨醇(DHS)、C16 DHS、C20 DHS、十八碳4-羟双氢(神经)鞘氨醇(C18PHS)、C20 PHS 11种AD血浆生物标记物,通过比较发现,AD模型组小鼠血浆中11种标记物含量较对照组均显著降低(P<0.01),而治疗组小鼠血浆中11种生物标记物含量较AD模型组均明显上升(P<0.05,P<0.01)。结论:黑水缬草提取物可通过干预AD小鼠体内氨基酸、磷脂和鞘脂等相关代谢通路而缓解小鼠海马区淀粉样斑块沉积,从而起到治疗AD作用。

Objective: To study the intervention mechanism of Valeriana amurensis extractive on Aβ_(25-35) induced Alzheimer's disease(AD) model mice by UPLC-MS technology. Methods: The experimental mice were randomly divided into three groups: control group, AD model group and treatment group. The pathological changes of brain tissue in micewere observed by immunohistochemistry. And UPLC-MS technique was conducted to study the regulation mechanism of Valeriana amurensis extract on biomarkers of AD mice.Results: The mice in control group showed that hippocampus cells arranged orderly and had normal morphology. Mice in AD model group showed a large number of amyloid plaques. Mice in treatment group showed decreased amyloid plaques compared to the AD mice. Eleven plasma biomarkers were identified by UPLC-MS, which were tryptophan, histidine, LPC(C22∶6, C18∶2, C16∶0), DHS, DHS(C16, C20), PHS(C18, C20). When compared to the control group, the levels of eleven kinds of plasma biomarkers in AD model group were significantly reduced(P<0.05, P<0.01). But after intervention of Valeriana amurensis extract, the contents of eleven kinds of markers in mice plasma were obvious increased(P<0.05, P<0.01). Conclusion: Valeriana amurensis extractive have a protective effect on AD mice, and its treatment mechanism may be closely related to interfering the related metabolic pathways such as amino acids, phospholipids and sphingolipids, so as to alleviate the deposition of amyloid plaques in hippocampus of mice.