胃复方干预SGC-7901荷瘤裸鼠的血浆非靶标代谢组学研究★

Non-target plasma metabolomics of Weifufang in SGC-7901 tumor-bearing nude mice★

摘要:

目的 利用超高效液相色谱-质谱（UHPLC-MS）非靶标技术探索胃复方干预SGC-7901荷瘤裸鼠的潜在代谢通路，为研究胃复方的体内代谢机制提供基础。方法 将胃癌细胞SGC-7901接种于6只裸鼠右侧乳垫处皮下，制备供瘤鼠模型。供瘤鼠瘤块长至10 mm×10 mm左右时，剥离瘤体进行鼠间传代接种，建立SGC-7901皮下移植瘤模型。将24只实验鼠按随机数字表法分为对照组和胃复方高、中、低剂量组，每组6只。对照组裸鼠每天给予0.2 mL生理盐水灌胃，胃复方高、中、低剂量组裸鼠每天给予0.2 mL相应药物灌胃，均连续给药14 d。通过祼鼠体质量、瘤体质量、抑瘤率分析干预效果，采用HE染色观察肿瘤标本。使用R程序包提取代谢物信息后，运用SIMCA软件进行主成分分析（PCA）及正交偏最小二乘法-判别分析（OPLS-DA），在Metabo Analyst软件中筛选出意义最大的代谢通路。结果 与对照组比较，中剂量胃复方抑制肿瘤生长效果较好（P<0.05），抑瘤率为35.97%，其次是高剂量和低剂量（P<0.05），抑瘤率分别为30.29%、30.07%。给药14 d内，对照组裸鼠体质量均高于胃复方高、中、低剂量组；第11天起，胃复方高、中剂量组裸鼠体质量均明显低于对照组（P<0.05）。胃复方干预荷瘤裸鼠后，筛选出19种显著差异代谢物，包括甜菜碱、葫芦巴碱、水苏碱、蛋氨酸亚砜、L-酪氨酸、2，3-二羟基-3-甲基丁酸酯、肌酸、鸟氨酸、L-脯氨酸、N-乙酰亮氨酸等，其中精氨酸和脯氨酸代谢通路，烟酸和烟酰胺代谢通路，苯丙氨酸、酪氨酸和色氨酸代谢通路为3条关键代谢通路。结论 胃复方能抑制SGC-7901荷瘤裸鼠肿瘤生长，可能与调节体内氨基酸代谢通路有关。

Abstract:

Objective To explore the potential metabolic pathway of Weifufang in the intervention of SGC-7901 tumor-bearing nude mice by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) non-target metabolomics technology, in order to provide a basis for the study of metabolic mechanism of Weifufang in vivo.Methods The SGC-7901 gastric cancer cells were inoculated subcutaneously into the right breast pad of 6 nude mice (donor mice) to construct the tumor donor mice model. When the tumor mass of donor mice grew to about 10 mm×10 mm, the tumor was dissected and subcultured to establish the SGC-7901 subcutaneous transplanted tumor model. 24 experimental nude mice were divided into control group, Weifufang high-dose group, medium-dose group, low-dose group, with 6 mice in each group. The control group was given 0.2 mL normal saline by intragastric administration every day. The Weifufang high-, medium-, low-dose groups were given by intragastric administration 0.2 mL Weifufang decoction in corresponding concentrations (52, 26, 13 g?kg^-1) for 14 consecutive days. The effect of intervention was analyzed by weight, tumor weight, tumor inhibition rate of nude mice, and tumor samples were observed by HE staining. R program package was used to extract metabolite information, and SIMCA software was used for principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). The most significant metabolic pathways were screened out with Metabo Analyst software.Results Compared with the control group, the Weifufang medium-dose group had better inhibitory effect on tumor growth (P<0.05), and the inhibitory rate was 35.97%, followed by the Weifufang high-dose group and low-dose group (P<0.05) with the inhibitory rates of 30.29% and 30.07% respectively. After 14 days of administration, the body weight of nude mice in the control group was higher than that in the Weifufang high-, medium- and low-dose groups. From the 11th day, the body weight of nude mice in the Weifufang high- and medium- dose groups was significantly lower than that in the control group (P<0.05). Totally, 19 significantly different metabolites were screened, including betaine, trigonelline, stachydrine, methionine sulfoxide, L-tyrosine, 2, 3-dihydroxy-3-methylbutanoate, creatine, ornithine, L-proline, N-acetylleucine. Three key metabolic pathways were the arginine and proline metabolic pathway, the nicotinic acid and nicotinamide metabolic pathway, and the phenylalanine, tyrosine and tryptophan metabolic pathway.Conclusion Weifufang can inhibit the tumor growth in SGC-7901 nude mice, which may be related with its regulation on the amino acids metabolism pathway in vivo.