阿纳其根醇提物对D-半乳糖致衰老小鼠抗衰老作用的血清核磁共振代谢组学研究

Study on anti-aging effect of Anacyclus pyrethrum ethanol extracts to D-galactose induced aging model mice by serum nuclear magnetic resonance metabolomics

目的利用核磁共振代谢组学方法研究阿纳其根提取物对D-半乳糖致衰老小鼠抗衰老作用机制。方法选取3月龄的昆明小鼠60只并将其随机分为正常组、模型组、阳性对照组和阿纳其根95%醇提物低、中、高剂量给药组,正常组以外的小鼠每天1次皮下注射D-半乳糖100mg/kg,同时对阿纳其根低、中、高剂量给药组分别灌胃相应浓度的阿纳其根95%醇提物(0.15、0.3、0.6g/kg),对阳性对照组灌胃盐酸多奈哌齐(0.92 mg/kg),模型组灌胃等体积的生理盐水,连续给药45 d,第45 d末次给药1 h后利用Morris水迷宫实验观察小鼠学习和记忆能力并收集各组小鼠血清,用1H-核磁共振氢谱法(hydrogen-nuclear magnetic resonance,1H-NMR)测定核磁共振氢谱,采用偏最小二乘回归分析法(partial least squares-discriminant analysis, PLS-DA)和正交偏最小二乘法判别分析法(orthogonalpartialleastsquares-discriminantanalysis,OPLS-DA)分析研究每组小鼠血清中代谢产物差异。结果行为学检测结果显示与模型组小鼠比较,阳性对照组和阿纳其根醇提物不同剂量组的逃避潜伏期逐渐缩短,差异有统计学意义(P<0.01);各组小鼠的空间探索实验显示与模型组相比,阿纳其根95%醇提物各剂量组小鼠经过有效区次数显著增加(P<0.05);且各组血清多种代谢物有显著性差异(P<0.05)。结论 D-半乳糖诱导的衰老模型小鼠的糖代谢代谢、氨基酸代谢发生紊乱,而阿纳其根醇提物对其紊乱有一定的改善作用从而发挥抗衰老作用。

Objective To study the anti-aging mechanism of Anacyclus pyrethrum root extracts on D-galactose-induced aging mice using nuclear magnetic resonance metabolomics method. Methods Sixty 3-month-old Kunming mice were randomly divided into normal group, model group, positive control group and low, medium and high-dose groups of 95% alcohol extract of Anacyclus pyrethrum. Mice outside the normal group were injected subcutaneously with D-galactose 100 mg/kg once a day. At the same time, Anacyclus pyrethrum roots low-, medium-, and high-dose groups were gave corresponding concentrations of Anacyclus pyrethrum root 95% alcohol extract(0.15, 0.3, 0.6 g/kg) by gavage. The positive control group was given donepezil hydrochloride(0.92 mg/kg) by gavage, and the model group was given an equal volume of normal saline by gavage. After continuous administration for 45 d, the learning and memory ability of mice were observed by Morris water maze experiment and serum of each group were collected after the last administration for 1 h on 45 d. Hydrogen-nuclear magnetic resonance spectroscopy was measured by hydrogen-nuclear magnetic resonance(1 H-NMR). The differences in serum metabolites of each group of mice were analyzed and studied by partial least squares-discriminant analysis(PLS-DA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Results The behavioral test results showed that compared with the model group of mice, the escape latency of the positive control group and the alcohol extract of Anacyclus pyrethrum root in different dose groups gradually shortened, and the differences were statistically significant. The space exploration experiment of each group of mice showed that compared with the model group, the number of times the mice in each dose group of Anacyclus pyrethrum 95% alcohol extract passed through the effective area was significantly increased(P<0.05). And there were significant differences in various serum metabolites in each group(P<0.05). Conclusion D-galactose-induced aging model mice had disorder of glucose metabolism and amino acid metabolism. Ethanol extracts of Anacyclus pyrethrum can significantly improve the learning and memory ability of aging mice, it`s anti-ageing effect may be achieved by regulating the disordered sugar and amino acid metabolism in the body of aging mice.