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基于LC-MS代谢组学研究黄芪注射液对白细胞减少症模型小鼠的干预作用
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| 篇名： | 基于LC-MS代谢组学研究黄芪注射液对白细胞减少症模型小鼠的干预作用 |
| TITLE： | Study on the Intervention Effects of Huangqi Injection on Leukopenia Model Mice Based on LC-MS Metabolomics |
| 摘要： | 目的：研究黄芪注射液对白细胞减少症模型小鼠的干预作用。方法：将昆明种小鼠随机分为正常组、模型组和药物组，每组8只。模型组和药物组小鼠均腹腔注射环磷酰胺以复制白细胞减少症模型，正常组小鼠腹腔注射等体积无菌注射用水。造模成功后，正常组和模型组小鼠均腹腔注射等体积无菌注射用水，药物组小鼠腹腔注射黄芪注射液0.04mL/10g，每日1次，连续7d。在检测各组小鼠血常规指标（白细胞、淋巴细胞、中性粒细胞、单核细胞计数）的基础上，采用液质联用技术（LC-MS）分析血清中的代谢产物，并通过主成分分析（PCA）、偏最小二乘法-判别分析（PLS-DA）、正交偏最小二乘法-判别分析（OPLS-DA）等多元统计分析以及HMDB、METLIN、KEGG等数据库和相关文献指认、鉴定差异代谢物；借助MetPA在线工具对差异代谢物的代谢通路进行分析，并基于皮尔逊相关系数分析血常规指标与差异代谢物的相关性。结果：与正常组比较，模型组小鼠血常规指标计数均显著减少（P＜0.01）；与模型组比较，黄芪注射液组小鼠上述血常规指标均显著增加（P＜0.01）。正常组和模型组小鼠血清样品的LC-MS图谱明显不同，且黄芪注射液组血清样品LC-MS代谢组学数据与正常组接近。多元统计分析和相关数据库分析显示，与正常组比较，模型组小鼠血清中L-异亮氨酸、二十碳五烯酸等17个代谢物含量显著升高，亚精胺等4个代谢物含量显著降低（P＜0.05或P＜0.01）；与模型组比较，黄芪注射液可回调小鼠血清中柠檬酸、L-脯氨酸、乙酰肉碱、L-异亮氨酸、L-苯丙氨酸、1-磷酸鞘氨醇、溶血磷脂酰肌醇、二十碳五烯酸和亚油酸等9个代谢物的含量（P＜0.05或P＜0.01），且主要与亚油酸代谢，苯丙氨酸、酪氨酸和色氨酸的生物合成，苯丙氨酸代谢等通路有关（代谢通路影响值均大于0.1）。相关性分析显示，血常规指数与模型组中含量升高的D-鞘氨醇、亚油酸、柠檬酸等含量之间的相关性较明显（r的绝对值普遍大于0.5）。结论：黄芪注射液可增加白细胞减少症模型小鼠的白细胞、淋巴细胞、中性粒细胞、单核细胞计数；其升高白细胞的作用可能与亚油酸代谢，苯丙氨酸、酪氨酸和色氨酸的生物合成及苯丙氨酸代谢等途径有关。 |
| ABSTRACT： | OBJECTIVE：To study the intervention effects of Huan gqi injecti on on leucopenia model mice. METHODS ： Kunming mice were randomly divided into normal group ，model group and drug group ，with 8 mice in each group. Model group and drug group were given intraperitoneal injection of cyclophosphamide to induce leukopenia model. Normal group was given intraperitoneal injection of equal volume of sterile water. After modeling successfully ，normal group and model group were given intraperitoneal injection of equal volume of sterile water ；drug group was given intraperitoneal injection of Huangqi injection 0.04 mL/10 g，once a day ，for consecutive 7 d. Based on the detection of blood routine indexes （leukocyte，lymphocyte，neutrophil， monocyte count ）of mice in each group ，the metabolites in serum were analyzed by LC-MS. Principal component analysis （PCA）， partial least squares discriminant analysis （PLS-DA），orthogonal partial least squares-discriminant analysis （OPLS-DA），HMDB， METLIN， KEGG and other databases as well as related literatures were used to identify the differential metabolites. The metabolic pathways of differential metabolites were analyzed with MetPA online tools ，and the correlation of blood routine indexes and differential metabolites were analyzed on the basis of Pearson correlation coefficient. RESULTS： Compared with normal group ， blood rountine indexes of model group were decreased significantly （P＜ 0.01）. Compared with model group ，above blood r ountine indexes of drug group were all increased siginificantly （P＜0.01）. LC-MS chromatogram of serum samples in normal group and model group were significantly different ，and LC-MS metabonomics data of serum samples in drug group were similar to those of normal group. Multivariate statistical analysis and correlated database analysis revealed that compared with normal group ，serum contents of 17 metabolites as L-isoleucine，eicosapentaenoic acid were increased significantly in model group ，while the contents of 4 metabolites as spermidine were decreased significantly （P＜0.05 or P＜0.01）. Compared with model group ，Huangqi injection could reverse the serum contents of 9 metabolites in mice ，such as citric acid ，L-proline，acetylcarnitine，L-isoleucine， L-phenylalanine，sphingosine-1-phosphate，lysophosphatidylinositol，eicosapentaenoic acid and linoleic acid （P＜0.05 or P＜ 0.01），which were associated with linoleic acid metabolism ，biosynthesis of phenylalanine ，tyrosine and tryptophan ，and phenylalanine metabolism （metabolism pathway influential values were all higher than 0.1）. Correlation analysis showed that there was a significant correlation between blood routine indexes and the contents of D-sphingosine，linoleic acid and citric acid in model group（the absolute values of r were generally greater than 0.5）. CONCLUSIONS ：Huangqi injection can increase the counts of leukocytes，lymphocytes，neutrophils and monocytes in leucopenia model mice. The increase of leukocytes may be related to linoleic acid metabolism ，biosynthesis of phenylalanine ，tyrosine and tryptophan ，and phenylalanine metabolism. |
| 期刊： | 2020年第31卷第21期 |
| 作者： | 刘腾，许晋芳，卢晓林，侯宏保，李震宇，曲婷丽，赵正保 |
| AUTHORS： | LIU Teng，XU Jinfang ，LU Xiaolin ，HOU Hongbao ，LI Zhenyu ，QU Tingli，ZHAO Zhengbao |
| 关键字： | 黄芪注射液；白细胞减少症；环磷酰胺；液质联用技术；代谢组学；小鼠 |
| KEYWORDS： | Huangqi injection ；Leucopenia；Cyclophosphamide；LC-MS；Metabolomics；Mice |
| 阅读数： | 394 次 |
| 本月下载数： | 3 次 |
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